Hot off the presses! Jun 01 Nat Med

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Latest Articles Include:

• When off label is off target
  - Nat Med 17(6):633 (2011)
  Nature Medicine | Editorial When off label is off target Journal name:Nature MedicineVolume: 17,Page:633Year published:(2011)DOI:doi:10.1038/nm0611-633Published online06 June 2011 Off-label prescribing of drugs can add to a clinician's toolbox, but it may also pose risks to patients. Careful monitoring and clinical validation should increase the safety and guarantee the efficacy of using existing drugs for new indications. View full text Additional data
• Market overlap points to irresponsible use of tuberculosis drugs
  - Nat Med 17(6):635 (2011)
  Article preview View full access options Nature Medicine | News Market overlap points to irresponsible use of tuberculosis drugs * Erika Check-HaydenJournal name:Nature MedicineVolume: 17,Page:635Year published:(2011)DOI:doi:10.1038/nm0611-635Published online06 June 2011 sy5/sy5/ZUMA Press/Newscom The private drug market can treat two thirds of those who develop TB. People with active tuberculosis infections turn to the private market for treatment far more often than anyone had realized. And when they do, they encounter a chaotic array of treatment choices, many of which do not meet guidelines drawn up by the World Health Organization. These are the conclusions of a paper published on 4 May that counters the prevailing wisdom that the vast majority of people with tuberculosis are treated through publicly funded programs. Article preview Read the full article * Instant access to this article: US$32 Buy now * Subscribe to Nature Medicine for full access: Subscribe * Personal subscribers: Log in Additional access options: * Login via Athens * Login via your Institution * Purchase a site license * Use a document delivery service * Rent this article from DeepDyve * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. Additional data Author Details * Erika Check-Hayden Search for this author in: * NPG journals * PubMed * Google Scholar
• New diabetes drugs go beyond insulin to flush out excess sugar
  - Nat Med 17(6):636 (2011)
  Article preview View full access options Nature Medicine | News New diabetes drugs go beyond insulin to flush out excess sugar * Monica HegerJournal name:Nature MedicineVolume: 17,Page:636Year published:(2011)DOI:doi:10.1038/nm0611-636aPublished online06 June 2011 The list of approved medications for type 2 diabetes is long and varied. Some drugs come in pill form; others must be injected. Some cause weight gain; others trigger weight loss. But all existing treatments to lower blood sugar, different as they may be, essentially work by modulating the activity of insulin. These drugs have proven quite effective in the short term, but they tend to lose their potency over time. As a result, drug companies have been racing to develop new diabetes medicines that act on a pathway that doesn't involve insulin. A drug called dapagliflozin could be the first such agent to gain market approval. Co-developed by Bristol-Myers Squibb (BMS) and AstraZeneca, dapagliflozin reduces blood glucose levels in an insulin-independent manner by preventing a protein called sodium-dependent glucose cotransporter 2 (SGLT2) from reabsorbing glucose in the kidney. As such, the simple sugar gets excreted through the urine instead of reentering the blood stream and elevating blood sugar levels, a condition known as hyperglycemia, one of the main symptoms of diabetes. Article preview Read the full article * Instant access to this article: US$32 Buy now * Subscribe to Nature Medicine for full access: Subscribe * Personal subscribers: Log in Additional access options: * Login via Athens * Login via your Institution * Purchase a site license * Use a document delivery service * Rent this article from DeepDyve * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. Additional data Author Details * Monica Heger Search for this author in: * NPG journals * PubMed * Google Scholar
• Disputed EU herbal medicine rules take force
  - Nat Med 17(6):636 (2011)
  Article preview View full access options Nature Medicine | News Disputed EU herbal medicine rules take force * Karen DenteJournal name:Nature MedicineVolume: 17,Page:636Year published:(2011)DOI:doi:10.1038/nm0611-636bPublished online06 June 2011 A controversial set of rules in the EU that changes the way herbal medicinal products are licensed and marketed in Europe came into full force this May, ending a seven-year grace period that had allowed manufacturers to come up to speed on the new measures. Under the law, any plant product with a new curative claim sold over the counter is subject to an approval process the same as the one that pharmaceutical drugs are required to undergo when seeking marketing authorization. Additionally the new regulations, known as the Traditional Herbal Medicinal Products Directive, will force natural products producers to adhere to the standards of good manufacturing practice, a set of guidelines designed to ensure the quality of a product. Article preview Read the full article * Instant access to this article: US$32 Buy now * Subscribe to Nature Medicine for full access: Subscribe * Personal subscribers: Log in Additional access options: * Login via Athens * Login via your Institution * Purchase a site license * Use a document delivery service * Rent this article from DeepDyve * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. Additional data Author Details * Karen Dente Search for this author in: * NPG journals * PubMed * Google Scholar
• HPV protection in older groups may be in the eye of the beholder
  - Nat Med 17(6):637 (2011)
  Article preview View full access options Nature Medicine | News New diabetes drugs go beyond insulin to flush out excess sugar * Monica HegerJournal name:Nature MedicineVolume: 17,Page:636Year published:(2011)DOI:doi:10.1038/nm0611-636aPublished online06 June 2011 The list of approved medications for type 2 diabetes is long and varied. Some drugs come in pill form; others must be injected. Some cause weight gain; others trigger weight loss. But all existing treatments to lower blood sugar, different as they may be, essentially work by modulating the activity of insulin. These drugs have proven quite effective in the short term, but they tend to lose their potency over time. As a result, drug companies have been racing to develop new diabetes medicines that act on a pathway that doesn't involve insulin. A drug called dapagliflozin could be the first such agent to gain market approval. Co-developed by Bristol-Myers Squibb (BMS) and AstraZeneca, dapagliflozin reduces blood glucose levels in an insulin-independent manner by preventing a protein called sodium-dependent glucose cotransporter 2 (SGLT2) from reabsorbing glucose in the kidney. As such, the simple sugar gets excreted through the urine instead of reentering the blood stream and elevating blood sugar levels, a condition known as hyperglycemia, one of the main symptoms of diabetes. Article preview Read the full article * Instant access to this article: US$32 Buy now * Subscribe to Nature Medicine for full access: Subscribe * Personal subscribers: Log in Additional access options: * Login via Athens * Login via your Institution * Purchase a site license * Use a document delivery service * Rent this article from DeepDyve * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. Additional data Author Details * Monica Heger Search for this author in: * NPG journals * PubMed * Google Scholar
• NIH funding rates drop to record lows
  - Nat Med 17(6):637 (2011)
  Article preview View full access options Nature Medicine | News Disputed EU herbal medicine rules take force * Karen DenteJournal name:Nature MedicineVolume: 17,Page:636Year published:(2011)DOI:doi:10.1038/nm0611-636bPublished online06 June 2011 A controversial set of rules in the EU that changes the way herbal medicinal products are licensed and marketed in Europe came into full force this May, ending a seven-year grace period that had allowed manufacturers to come up to speed on the new measures. Under the law, any plant product with a new curative claim sold over the counter is subject to an approval process the same as the one that pharmaceutical drugs are required to undergo when seeking marketing authorization. Additionally the new regulations, known as the Traditional Herbal Medicinal Products Directive, will force natural products producers to adhere to the standards of good manufacturing practice, a set of guidelines designed to ensure the quality of a product. Article preview Read the full article * Instant access to this article: US$32 Buy now * Subscribe to Nature Medicine for full access: Subscribe * Personal subscribers: Log in Additional access options: * Login via Athens * Login via your Institution * Purchase a site license * Use a document delivery service * Rent this article from DeepDyve * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. Additional data Author Details * Karen Dente Search for this author in: * NPG journals * PubMed * Google Scholar
• Gene tests for brain injury still far from the football field
  - Nat Med 17(6):638 (2011)
  Article preview View full access options Nature Medicine | News Market overlap points to irresponsible use of tuberculosis drugs * Erika Check-HaydenJournal name:Nature MedicineVolume: 17,Page:635Year published:(2011)DOI:doi:10.1038/nm0611-635Published online06 June 2011 sy5/sy5/ZUMA Press/Newscom The private drug market can treat two thirds of those who develop TB. People with active tuberculosis infections turn to the private market for treatment far more often than anyone had realized. And when they do, they encounter a chaotic array of treatment choices, many of which do not meet guidelines drawn up by the World Health Organization. These are the conclusions of a paper published on 4 May that counters the prevailing wisdom that the vast majority of people with tuberculosis are treated through publicly funded programs. Article preview Read the full article * Instant access to this article: US$32 Buy now * Subscribe to Nature Medicine for full access: Subscribe * Personal subscribers: Log in Additional access options: * Login via Athens * Login via your Institution * Purchase a site license * Use a document delivery service * Rent this article from DeepDyve * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. Additional data Author Details * Erika Check-Hayden Search for this author in: * NPG journals * PubMed * Google Scholar
• Jewish genetic screening grows despite questions about breadth
  - Nat Med 17(6):639 (2011)
  Article preview View full access options Nature Medicine | News Market overlap points to irresponsible use of tuberculosis drugs * Erika Check-HaydenJournal name:Nature MedicineVolume: 17,Page:635Year published:(2011)DOI:doi:10.1038/nm0611-635Published online06 June 2011 sy5/sy5/ZUMA Press/Newscom The private drug market can treat two thirds of those who develop TB. People with active tuberculosis infections turn to the private market for treatment far more often than anyone had realized. And when they do, they encounter a chaotic array of treatment choices, many of which do not meet guidelines drawn up by the World Health Organization. These are the conclusions of a paper published on 4 May that counters the prevailing wisdom that the vast majority of people with tuberculosis are treated through publicly funded programs. Article preview Read the full article * Instant access to this article: US$32 Buy now * Subscribe to Nature Medicine for full access: Subscribe * Personal subscribers: Log in Additional access options: * Login via Athens * Login via your Institution * Purchase a site license * Use a document delivery service * Rent this article from DeepDyve * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. Additional data Author Details * Erika Check-Hayden Search for this author in: * NPG journals * PubMed * Google Scholar
• Attending conferences... under the influence
  - Nat Med 17(6):640-641 (2011)
  Article preview View full access options Nature Medicine | News Market overlap points to irresponsible use of tuberculosis drugs * Erika Check-HaydenJournal name:Nature MedicineVolume: 17,Page:635Year published:(2011)DOI:doi:10.1038/nm0611-635Published online06 June 2011 sy5/sy5/ZUMA Press/Newscom The private drug market can treat two thirds of those who develop TB. People with active tuberculosis infections turn to the private market for treatment far more often than anyone had realized. And when they do, they encounter a chaotic array of treatment choices, many of which do not meet guidelines drawn up by the World Health Organization. These are the conclusions of a paper published on 4 May that counters the prevailing wisdom that the vast majority of people with tuberculosis are treated through publicly funded programs. Article preview Read the full article * Instant access to this article: US$32 Buy now * Subscribe to Nature Medicine for full access: Subscribe * Personal subscribers: Log in Additional access options: * Login via Athens * Login via your Institution * Purchase a site license * Use a document delivery service * Rent this article from DeepDyve * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. Additional data Author Details * Erika Check-Hayden Search for this author in: * NPG journals * PubMed * Google Scholar
• Chile plans new agency to boost pharma oversight
  - Nat Med 17(6):642 (2011)
  Article preview View full access options Nature Medicine | News New diabetes drugs go beyond insulin to flush out excess sugar * Monica HegerJournal name:Nature MedicineVolume: 17,Page:636Year published:(2011)DOI:doi:10.1038/nm0611-636aPublished online06 June 2011 The list of approved medications for type 2 diabetes is long and varied. Some drugs come in pill form; others must be injected. Some cause weight gain; others trigger weight loss. But all existing treatments to lower blood sugar, different as they may be, essentially work by modulating the activity of insulin. These drugs have proven quite effective in the short term, but they tend to lose their potency over time. As a result, drug companies have been racing to develop new diabetes medicines that act on a pathway that doesn't involve insulin. A drug called dapagliflozin could be the first such agent to gain market approval. Co-developed by Bristol-Myers Squibb (BMS) and AstraZeneca, dapagliflozin reduces blood glucose levels in an insulin-independent manner by preventing a protein called sodium-dependent glucose cotransporter 2 (SGLT2) from reabsorbing glucose in the kidney. As such, the simple sugar gets excreted through the urine instead of reentering the blood stream and elevating blood sugar levels, a condition known as hyperglycemia, one of the main symptoms of diabetes. Article preview Read the full article * Instant access to this article: US$32 Buy now * Subscribe to Nature Medicine for full access: Subscribe * Personal subscribers: Log in Additional access options: * Login via Athens * Login via your Institution * Purchase a site license * Use a document delivery service * Rent this article from DeepDyve * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. Additional data Author Details * Monica Heger Search for this author in: * NPG journals * PubMed * Google Scholar
• Drug companies hope to breathe life into asthma pipeline
  - Nat Med 17(6):642-643 (2011)
  Article preview View full access options Nature Medicine | News Disputed EU herbal medicine rules take force * Karen DenteJournal name:Nature MedicineVolume: 17,Page:636Year published:(2011)DOI:doi:10.1038/nm0611-636bPublished online06 June 2011 A controversial set of rules in the EU that changes the way herbal medicinal products are licensed and marketed in Europe came into full force this May, ending a seven-year grace period that had allowed manufacturers to come up to speed on the new measures. Under the law, any plant product with a new curative claim sold over the counter is subject to an approval process the same as the one that pharmaceutical drugs are required to undergo when seeking marketing authorization. Additionally the new regulations, known as the Traditional Herbal Medicinal Products Directive, will force natural products producers to adhere to the standards of good manufacturing practice, a set of guidelines designed to ensure the quality of a product. Article preview Read the full article * Instant access to this article: US$32 Buy now * Subscribe to Nature Medicine for full access: Subscribe * Personal subscribers: Log in Additional access options: * Login via Athens * Login via your Institution * Purchase a site license * Use a document delivery service * Rent this article from DeepDyve * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. Additional data Author Details * Karen Dente Search for this author in: * NPG journals * PubMed * Google Scholar
• On thirtieth anniversary, calls for HIV cure research intensify
  - Nat Med 17(6):643 (2011)
  Article preview View full access options Nature Medicine | News Market overlap points to irresponsible use of tuberculosis drugs * Erika Check-HaydenJournal name:Nature MedicineVolume: 17,Page:635Year published:(2011)DOI:doi:10.1038/nm0611-635Published online06 June 2011 sy5/sy5/ZUMA Press/Newscom The private drug market can treat two thirds of those who develop TB. People with active tuberculosis infections turn to the private market for treatment far more often than anyone had realized. And when they do, they encounter a chaotic array of treatment choices, many of which do not meet guidelines drawn up by the World Health Organization. These are the conclusions of a paper published on 4 May that counters the prevailing wisdom that the vast majority of people with tuberculosis are treated through publicly funded programs. Article preview Read the full article * Instant access to this article: US$32 Buy now * Subscribe to Nature Medicine for full access: Subscribe * Personal subscribers: Log in Additional access options: * Login via Athens * Login via your Institution * Purchase a site license * Use a document delivery service * Rent this article from DeepDyve * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. Additional data Author Details * Erika Check-Hayden Search for this author in: * NPG journals * PubMed * Google Scholar
• News in brief: Biomedical briefing
  - Nat Med 17(6):644-645 (2011)
  Article preview View full access options Nature Medicine | News Market overlap points to irresponsible use of tuberculosis drugs * Erika Check-HaydenJournal name:Nature MedicineVolume: 17,Page:635Year published:(2011)DOI:doi:10.1038/nm0611-635Published online06 June 2011 sy5/sy5/ZUMA Press/Newscom The private drug market can treat two thirds of those who develop TB. People with active tuberculosis infections turn to the private market for treatment far more often than anyone had realized. And when they do, they encounter a chaotic array of treatment choices, many of which do not meet guidelines drawn up by the World Health Organization. These are the conclusions of a paper published on 4 May that counters the prevailing wisdom that the vast majority of people with tuberculosis are treated through publicly funded programs. Article preview Read the full article * Instant access to this article: US$32 Buy now * Subscribe to Nature Medicine for full access: Subscribe * Personal subscribers: Log in Additional access options: * Login via Athens * Login via your Institution * Purchase a site license * Use a document delivery service * Rent this article from DeepDyve * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. Additional data Author Details * Erika Check-Hayden Search for this author in: * NPG journals * PubMed * Google Scholar
• Heat shock and awe
  - Nat Med 17(6):646-649 (2011)
  Nature Medicine | Editorial When off label is off target Journal name:Nature MedicineVolume: 17,Page:633Year published:(2011)DOI:doi:10.1038/nm0611-633Published online06 June 2011 Off-label prescribing of drugs can add to a clinician's toolbox, but it may also pose risks to patients. Careful monitoring and clinical validation should increase the safety and guarantee the efficacy of using existing drugs for new indications. View full text Additional data
• Caught on film
  - Nat Med 17(6):650-653 (2011)
  Nature Medicine | Editorial When off label is off target Journal name:Nature MedicineVolume: 17,Page:633Year published:(2011)DOI:doi:10.1038/nm0611-633Published online06 June 2011 Off-label prescribing of drugs can add to a clinician's toolbox, but it may also pose risks to patients. Careful monitoring and clinical validation should increase the safety and guarantee the efficacy of using existing drugs for new indications. View full text Additional data
• Persistence may pay off for antibiotics innovators
  - Nat Med 17(6):652 (2011)
  Article preview View full access options Nature Medicine | News Market overlap points to irresponsible use of tuberculosis drugs * Erika Check-HaydenJournal name:Nature MedicineVolume: 17,Page:635Year published:(2011)DOI:doi:10.1038/nm0611-635Published online06 June 2011 sy5/sy5/ZUMA Press/Newscom The private drug market can treat two thirds of those who develop TB. People with active tuberculosis infections turn to the private market for treatment far more often than anyone had realized. And when they do, they encounter a chaotic array of treatment choices, many of which do not meet guidelines drawn up by the World Health Organization. These are the conclusions of a paper published on 4 May that counters the prevailing wisdom that the vast majority of people with tuberculosis are treated through publicly funded programs. Article preview Read the full article * Instant access to this article: US$32 Buy now * Subscribe to Nature Medicine for full access: Subscribe * Personal subscribers: Log in Additional access options: * Login via Athens * Login via your Institution * Purchase a site license * Use a document delivery service * Rent this article from DeepDyve * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. Additional data Author Details * Erika Check-Hayden Search for this author in: * NPG journals * PubMed * Google Scholar
• Straight talk with...Joel Kupersmith
  - Nat Med 17(6):654 (2011)
  Nature Medicine | Editorial When off label is off target Journal name:Nature MedicineVolume: 17,Page:633Year published:(2011)DOI:doi:10.1038/nm0611-633Published online06 June 2011 Off-label prescribing of drugs can add to a clinician's toolbox, but it may also pose risks to patients. Careful monitoring and clinical validation should increase the safety and guarantee the efficacy of using existing drugs for new indications. View full text Additional data
• China needs to boost funding for graduate students to stay competitive
  - Nat Med 17(6):655 (2011)
  Nature Medicine | Editorial When off label is off target Journal name:Nature MedicineVolume: 17,Page:633Year published:(2011)DOI:doi:10.1038/nm0611-633Published online06 June 2011 Off-label prescribing of drugs can add to a clinician's toolbox, but it may also pose risks to patients. Careful monitoring and clinical validation should increase the safety and guarantee the efficacy of using existing drugs for new indications. View full text Additional data
• Vaccine politics
  - Nat Med 17(6):656 (2011)
  Article preview View full access options Nature Medicine | News Market overlap points to irresponsible use of tuberculosis drugs * Erika Check-HaydenJournal name:Nature MedicineVolume: 17,Page:635Year published:(2011)DOI:doi:10.1038/nm0611-635Published online06 June 2011 sy5/sy5/ZUMA Press/Newscom The private drug market can treat two thirds of those who develop TB. People with active tuberculosis infections turn to the private market for treatment far more often than anyone had realized. And when they do, they encounter a chaotic array of treatment choices, many of which do not meet guidelines drawn up by the World Health Organization. These are the conclusions of a paper published on 4 May that counters the prevailing wisdom that the vast majority of people with tuberculosis are treated through publicly funded programs. Article preview Read the full article * Instant access to this article: US$32 Buy now * Subscribe to Nature Medicine for full access: Subscribe * Personal subscribers: Log in Additional access options: * Login via Athens * Login via your Institution * Purchase a site license * Use a document delivery service * Rent this article from DeepDyve * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. Additional data Author Details * Erika Check-Hayden Search for this author in: * NPG journals * PubMed * Google Scholar
• The holy grail of high bone mass
  - Nat Med 17(6):657-658 (2011)
  Article preview View full access options Nature Medicine | Article Lrp5 functions in bone to regulate bone mass * Yajun Cui1, 2 * Paul J Niziolek3, 4, 5 * Bryan T MacDonald6, 7 * Cassandra R Zylstra8 * Natalia Alenina9 * Daniel R Robinson8 * Zhendong Zhong8 * Susann Matthes9 * Christina M Jacobsen1 * Ronald A Conlon2 * Robert Brommage10 * Qingyun Liu10 * Faika Mseeh10 * David R Powell10 * Qi M Yang10 * Brian Zambrowicz10 * Han Gerrits11 * Jan A Gossen11 * Xi He6, 7 * Michael Bader9 * Bart O Williams8 * Matthew L Warman1, 12, 13 * Alexander G Robling4, 5 * Affiliations * Contributions * Corresponding authorJournal name:Nature MedicineVolume: 17,Pages:684–691Year published:(2011)DOI:doi:10.1038/nm.2388Received14 December 2010Accepted27 April 2011Published online22 May 2011 Abstract * Abstract * Author information * Supplementary information Article tools * Full text * 日本語要約 * Print * Email * Download citation * Order reprints * Rights and permissions * Share/bookmark * Connotea * CiteULike * Facebook * Twitter * Delicious * Digg The human skeleton is affected by mutations in low-density lipoprotein receptor-related protein 5 (LRP5). To understand how LRP5 influences bone properties, we generated mice with osteocyte-specific expression of inducible Lrp5 mutations that cause high and low bone mass phenotypes in humans. We found that bone properties in these mice were comparable to bone properties in mice with inherited mutations. We also induced an Lrp5 mutation in cells that form the appendicular skeleton but not in cells that form the axial skeleton; we observed that bone properties were altered in the limb but not in the spine. These data indicate that Lrp5 signaling functions locally, and they suggest that increasing LRP5 signaling in mature bone cells may be a strategy for treating human disorders associated with low bone mass, such as osteoporosis. Figures at a glance * Figure 1: Generation and characterization of HBM-causing Lrp5 knock-in mice. () Schematic depicting the targeting and genotyping strategies for Lrp5 HBM alleles. The 5′ targeting arm contains a neomycin-resistance cassette (NeoR) flanked by loxP sites (arrowheads). The 3′ targeting arm begins in intron 2 and extends into intron 4, and is followed by a thymidine kinase (TK) cassette. Site-directed mutagenesis altered specific amino acid residues encoded by Lrp5 exon 3 (asterisks). The relative locations and orientation of primers used for PCR genotyping and their expected amplimer sizes are noted. () Autoradiographs of a northern blot containing whole-bone total RNA from mice with different Lrp5 genotypes initially hybridized with a radioactive Lrp5 cDNA probe (top) and subsequently with a glyceraldehyde 3-phosphate dehydrogenase (Gapdh) cDNA probe which serves as a loading control (bottom). () Photograph of an agarose gel containing PCR amplimers derived from genomic DNA of mice with different Lrp5 genotypes. () Graphs depicting the areal bone mi! neral density (aBMD) measured by dual-energy X-ray absorptiometry (DEXA) in mice with different Lrp5 genotypes; mice followed until the age of 16.5 weeks (top and middle). Graphs depicting the percentage of trabecular bone volume in the total volume (BV/TV) of the distal femora and fifth lumbar vertebrae of 16.5-week-old male and female mice with different Lrp5 genotypes (bottom). () Representative μCT scan images obtained from 16.5-week-old mice with different Lrp5 genotypes. Scale bars, 1 mm. () Graphs depicting biomechanical properties of whole femora in a three-point bending assay from 16.5-week-old mice with different Lrp5 genotypes. () Representative images of new bone formation assessed by double calcein labeling of mice with different Lrp5 genotypes. Bone formation rates/bone surface area (BFR/BS); mineral apposition rates (MAR); mineralizing surface/bone surface (MS/BS). The numbers of mice studied are indicated. Error bars show means ± s.d. *P < 0.05 versus WT m! ice; #P < 0.05 versus heterozygous Lrp5 HBM mice. * Figure 2: Effect of activating Lrp5 NeoR-containing HBM alleles. () Graphs depicting femoral and vertebral trabecular BV/TV in WT mice and in mice with Lrp5 NeoR-containing HBM alleles. (,) Graphs depicting femoral trabecular BV/TV () and vertebral trabecular BV/TV () in mice with (shaded bars) and without (unshaded bars) inherited NeoR-containing Lrp5 HBM alleles (+/AN or +/GN, and +/A or +/G, respectively), and with and without Vil1-Cre (+V and –V, respectively) or Dmp1-Cre (+D and –D, respectively) transgenes. () Photographs of agarose gels containing PCR amplimers derived from mouse genomic DNA extracted from either duodenum or femur cortex of mice with different Lrp5 and Cre-transgene genotypes. PCR amplimers correspond to the sizes depicted in Figure 1a. Top, amplimers from the Dmp1-Cre cross. Bottom, amplimers from the Vil1-Cre cross. WT allele (arrowheads), AN or GN allele (double arrowheads) A or G allele (arrows). () Graphs depicting fluorochrome-derived bone formation parameters in the distal femur from 9-week-old female mi! ce that were administered double calcein labeling. Group notations (x axis) follow those described for panel . () Graphs depicting the proportion of distal femur trabecular bone surface covered by osteoclasts (Oc.S; left) and osteoblasts (Ob.S; right). () Graphs depicting femoral and vertebral trabecular BV/TV in 12-week-old mice with (shaded bars) and without (unshaded bars) inherited NeoR-containing Lrp5 HBM alleles (+/AN and +/A), respectively, and with and without the Prrx1-Cre transgene (+P and −P, respectively). MS/BS, MAR, BFR/BS are as defined in Figure 1. The numbers of mice studied are indicated. Error bars show means ± s.d. *P < 0.05 versus NeoR-containing littermates that did not inherit a Cre transgene. * Figure 3: Generation and characterization of mice with a conditional knockout allele of Lrp5. () Schematic depicting the creation of the Lrp5 floxed allele. loxP sites (arrowheads), the neomycin-resistance cassette (NeoR), flippase (Flp) recognition target (FRT) sites (diamonds) and the diphtheria toxin (DT) cassette are shown. The relative locations and orientation of the three primers (arrows) used for PCR genotyping and their expected amplimer sizes are noted. () Photograph of agarose gel depicting PCR amplimers for WT (+), floxed (f) and knockout (−) Lrp5 alleles from genomic DNA of mice with different Lrp5 genotypes. () Photographs of agarose gels containing PCR amplimers derived from mouse genomic DNA extracted from either duodenum or femur cortex of floxed Lrp5 mice, with or without the Dmp1-Cre transgene (top) and with or without the Vil1-Cre transgene (bottom). () Graphs depicting whole-body aBMD (left), femoral trabecular BV/TV (middle) and vertebral trabecular BV/TV (right) in 16-week-old mice homozygous for WT or floxed Lrp5 alleles, with or without the! Dmp1-Cre transgene (+D and –D, respectively). () Graphs depicting whole-body aBMD and tibial trabecular BV/TV in 3-month-old and 12-month-old mice heterozygous or homozygous for floxed Lrp5 alleles with or without the Vil1-Cre transgene (+V and –V, respectively). The numbers of mice studied are indicated. Error bars show means ± s.d. *P < 0.05 versus floxed Lrp5 littermates that did not inherit the Cre transgene. * Figure 4: Effect of Lrp5 genotype on 5HT concentration and on Tph1 expression. () Graphs depicting whole-blood 5HT measured by HPLC in 6-month-old Lrp5 WT and knockout mice that had been backcrossed to C57BL6/J mice. Shown are 5HT measurements in Lrp5 WT and HBM-causing knock-in (G/G) mice on a mixed 129Sv/C57BL/6J background (far left); in 3-month-old Lrp5 WT, knockout and HBM knock-in (+/A) mice on a mixed 129Sv/C57BL/6J genetic background (middle left); and in 3-month-old male (middle right) and 13-month-old female (far right) WT and knockout littermates on a mixed 129SvEvBrd/ C57BL/6J-Tyrc-Brd background. () Graphs depicting the quantity of 5HT extracted from several regions of the intestine, beginning in the duodenum and proceeding through the jejunum, ileum and proximal colon in 3-month-old male (left) and in 13-month-old female (right) Lrp5 WT and knockout littermates on a mixed 129SvEvBrd/ C57BL/6J-Tyrc-Brd background. () Scattergram depicting vertebral trabecular BV/TV and whole blood 5HT measurements in individual Lrp5 WT (open symbols) and k! nockout (filled symbols) littermates. Correlations between BV/TV and whole blood serotonin were r2 = 0.13 (P = 0.16) for male mice, and r2 = 0.02 (P = 0.53) for female mice. () Graphs depicting normalized Tph1 transcript levels in duodenum RNA extracts from Lrp5 WT, knockout, and HBM-causing knock-in (+/A) mice on a mixed 129Sv/C57BL/6J genetic background (left) and duodenum and colon RNA extracts from Lrp5 WT and knockout mice on a 129SvEvBrd/ C57BL/6J-Tyrc-Brd background (right) with Gapdh serving as the internal control. The mean Tph1 expression level for Lrp5 WT duodenum is set as 100%. The numbers of mice studied are indicated. Error bars show means ± s.d. *P < 0.05 versus WT mice. * Figure 5: Bone mass in WT and Tph1−/− mice. () Graphs depicting femoral trabecular BV/TV in WT and Tph1−/− mice on either FVB/N or C57BL/6 backgrounds (left) or on a mixed 129SvEvBrd/C57BL/6J-Tyrc-Brd background (right). () Graphs depicting the vertebral trabecular BV/TV of the fifth lumbar vertebrae in the same mice described in panel . () Graphs depicting lumbar spine aBMD, as measured by DEXA, in the same mice described in panel . The numbers of mice studied are indicated. Error bars show means ± s.d. *P < 0.05 versus WT mice using an unpaired t test; none of these differences remain significant after correcting for multiple testing. * Figure 6: Bone mass after pharmacologic inhibition of Tph1 activity. () Graph depicting dose-dependent changes in 5HT content, compared to vehicle-treated controls, in 9-week-old WT female C57BL/6 mice after receiving daily doses of LP-923941 for 7 days. A daily dose of 250 mg kg−1 lowered 5HT content in whole blood and in intestine, but not in brain. () Graph depicting changes in the intestinal 5HT content in sham-operated (SHM) and ovariectomized (OVX) mice that received vehicle or LP-923941 (250 mg per kg per day) for 6 weeks. Ovariectomy alone reduced 5HT content in the duodenum and colon by ~12% (P < 0.05) compared with SHM mice. Treatment with LP-923941 significantly reduced serotonin content equally in all regions of the intestine in SHM and in OVX mice. () Effect of treatment with LP-923941 (250 mg per kg per day) or teriparatide (80 μg per kg per day), which is the 1–34 residue amino-terminal fragment of human parathyroid hormone (PTH), on serum P1NP levels (left), vertebral trabecular BV/TV (middle left), midshaft femur cortica! l thickness (Ct.th; middle right) and midshaft femoral volumetric BMD (right) in SHM and in OVX mice. () Effect of treating SHM and OVX rats with LP-923941 (50 or 250 mg per kg per day) or teriparatide (80 μg per kg per day) for 6 weeks on jejunal 5HT content (left), femoral trabecular BV/TV (middle) and vertebral trabecular BV/TV (right). The numbers of mice studied are indicated. Error bars show means ± s.d. *P < 0.05 versus vehicle-treated mice. Article preview Read the full article * Instant access to this article: US$32 Buy now * Subscribe to Nature Medicine for full access: Subscribe * Personal subscribers: Log in Additional access options: * Login via Athens * Login via your Institution * Purchase a site license * Use a document delivery service * Rent this article from DeepDyve * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. Author information * Abstract * Author information * Supplementary information Affiliations * Orthopedic Research Laboratories, Department of Orthopedic Surgery, Children's Hospital, Boston, Massachusetts, USA. * Yajun Cui, * Christina M Jacobsen & * Matthew L Warman * Department of Genetics, Case Western Reserve University School of Medicine, Cleveland, Ohio, USA. * Yajun Cui & * Ronald A Conlon * Weldon School of Biomedical Engineering, Purdue University, West Lafayette, Indiana, USA. * Paul J Niziolek * Department of Anatomy & Cell Biology, Indiana University School of Medicine, Indianapolis, Indiana, USA. * Paul J Niziolek & * Alexander G Robling * Department of Biomedical Engineering, Indiana University School of Medicine, Indianapolis, Indiana, USA. * Paul J Niziolek & * Alexander G Robling * F.M. Kirby Neurobiology Center, Children's Hospital, Boston, Massachusetts, USA. * Bryan T MacDonald & * Xi He * Department of Neurology, Harvard Medical School, Boston, Massachusetts, USA. * Bryan T MacDonald & * Xi He * Center for Skeletal Disease Research, Van Andel Research Institute, Grand Rapids, Michigan, USA. * Cassandra R Zylstra, * Daniel R Robinson, * Zhendong Zhong & * Bart O Williams * Max Delbrück Center for Molecular Medicine, Berlin-Buch, Germany. * Natalia Alenina, * Susann Matthes & * Michael Bader * Lexicon Pharmaceuticals, The Woodlands, Texas, USA. * Robert Brommage, * Qingyun Liu, * Faika Mseeh, * David R Powell, * Qi M Yang & * Brian Zambrowicz * Merck Sharp & Dohme Research Laboratories, Oss, The Netherlands. * Han Gerrits & * Jan A Gossen * Howard Hughes Medical Institute, Children's Hospital, Boston, Massachusetts, USA. * Matthew L Warman * Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA. * Matthew L Warman Contributions Y.C. created and did studies on the mice with the Lrp5 HBM alleles and measured serum serotonin levels by competitive ELISA. P.J.N. did radiographic imaging and biomechanical testing on the mice with HBM-associated alleles. B.T.M. contributed to the serotonin and Tph1 qRT-PCR measurements in HBM-causing and Lrp5 knockout mice. C.R.Z. did multiple studies using the conditional Lrp5 knockout mice. N.A. studied the Tph1−/− mice, and with S.M. measured whole blood serotonin levels from HBM-causing and Lrp5-knockout mice by HPLC. D.R.R. generated the conditional Lrp5 knockout strain and Z.Z. participated in conditional inactivation of this allele using different Cre transgenes. C.M.J. carried out the Prrx1-Cre experiments. R.B., F.M. and Q.M.Y. organized studies on Lrp5- and Tph -knockout mice, and also organized the mouse pharmacology experiment. H.G. and J.A.G. organized the rat pharmacology experiment. R.A.C., X.H., M.B., D.R.P., Q.L., B.Z., B.O.W., A.G.R. and M.L.W. desig! ned experiments and provided reagents and financial support. M.L.W. prepared the first draft of the manuscript. All co-authors contributed detailed methods and results, and revised and approved the manuscript. Competing financial interests Employees of Lexicon Pharmaceuticals (R.B., Q.L., F.M., D.R.P., Q.M.Y. and B.Z.) and Merck Sharp & Dohme Research Laboratories (H.G. and J.A.G.) have received compensation in the form of salary and stock options. Corresponding author Correspondence to: * Matthew L Warman Author Details * Yajun Cui Search for this author in: * NPG journals * PubMed * Google Scholar * Paul J Niziolek Search for this author in: * NPG journals * PubMed * Google Scholar * Bryan T MacDonald Search for this author in: * NPG journals * PubMed * Google Scholar * Cassandra R Zylstra Search for this author in: * NPG journals * PubMed * Google Scholar * Natalia Alenina Search for this author in: * NPG journals * PubMed * Google Scholar * Daniel R Robinson Search for this author in: * NPG journals * PubMed * Google Scholar * Zhendong Zhong Search for this author in: * NPG journals * PubMed * Google Scholar * Susann Matthes Search for this author in: * NPG journals * PubMed * Google Scholar * Christina M Jacobsen Search for this author in: * NPG journals * PubMed * Google Scholar * Ronald A Conlon Search for this author in: * NPG journals * PubMed * Google Scholar * Robert Brommage Search for this author in: * NPG journals * PubMed * Google Scholar * Qingyun Liu Search for this author in: * NPG journals * PubMed * Google Scholar * Faika Mseeh Search for this author in: * NPG journals * PubMed * Google Scholar * David R Powell Search for this author in: * NPG journals * PubMed * Google Scholar * Qi M Yang Search for this author in: * NPG journals * PubMed * Google Scholar * Brian Zambrowicz Search for this author in: * NPG journals * PubMed * Google Scholar * Han Gerrits Search for this author in: * NPG journals * PubMed * Google Scholar * Jan A Gossen Search for this author in: * NPG journals * PubMed * Google Scholar * Xi He Search for this author in: * NPG journals * PubMed * Google Scholar * Michael Bader Search for this author in: * NPG journals * PubMed * Google Scholar * Bart O Williams Search for this author in: * NPG journals * PubMed * Google Scholar * Matthew L Warman Contact Matthew L Warman Search for this author in: * NPG journals * PubMed * Google Scholar * Alexander G Robling Search for this author in: * NPG journals * PubMed * Google Scholar Supplementary information * Abstract * Author information * Supplementary information PDF files * Supplementary Text and Figures (9M) Supplementary Figures 1–10, Supplementary Tables 1–4 and Supplementary Methods Additional data
• A CUE hints at tumor resistance
  - Nat Med 17(6):658-660 (2011)
  Article preview View full access options Nature Medicine | Article Lrp5 functions in bone to regulate bone mass * Yajun Cui1, 2 * Paul J Niziolek3, 4, 5 * Bryan T MacDonald6, 7 * Cassandra R Zylstra8 * Natalia Alenina9 * Daniel R Robinson8 * Zhendong Zhong8 * Susann Matthes9 * Christina M Jacobsen1 * Ronald A Conlon2 * Robert Brommage10 * Qingyun Liu10 * Faika Mseeh10 * David R Powell10 * Qi M Yang10 * Brian Zambrowicz10 * Han Gerrits11 * Jan A Gossen11 * Xi He6, 7 * Michael Bader9 * Bart O Williams8 * Matthew L Warman1, 12, 13 * Alexander G Robling4, 5 * Affiliations * Contributions * Corresponding authorJournal name:Nature MedicineVolume: 17,Pages:684–691Year published:(2011)DOI:doi:10.1038/nm.2388Received14 December 2010Accepted27 April 2011Published online22 May 2011 Abstract * Abstract * Author information * Supplementary information Article tools * Full text * 日本語要約 * Print * Email * Download citation * Order reprints * Rights and permissions * Share/bookmark * Connotea * CiteULike * Facebook * Twitter * Delicious * Digg The human skeleton is affected by mutations in low-density lipoprotein receptor-related protein 5 (LRP5). To understand how LRP5 influences bone properties, we generated mice with osteocyte-specific expression of inducible Lrp5 mutations that cause high and low bone mass phenotypes in humans. We found that bone properties in these mice were comparable to bone properties in mice with inherited mutations. We also induced an Lrp5 mutation in cells that form the appendicular skeleton but not in cells that form the axial skeleton; we observed that bone properties were altered in the limb but not in the spine. These data indicate that Lrp5 signaling functions locally, and they suggest that increasing LRP5 signaling in mature bone cells may be a strategy for treating human disorders associated with low bone mass, such as osteoporosis. Figures at a glance * Figure 1: Generation and characterization of HBM-causing Lrp5 knock-in mice. () Schematic depicting the targeting and genotyping strategies for Lrp5 HBM alleles. The 5′ targeting arm contains a neomycin-resistance cassette (NeoR) flanked by loxP sites (arrowheads). The 3′ targeting arm begins in intron 2 and extends into intron 4, and is followed by a thymidine kinase (TK) cassette. Site-directed mutagenesis altered specific amino acid residues encoded by Lrp5 exon 3 (asterisks). The relative locations and orientation of primers used for PCR genotyping and their expected amplimer sizes are noted. () Autoradiographs of a northern blot containing whole-bone total RNA from mice with different Lrp5 genotypes initially hybridized with a radioactive Lrp5 cDNA probe (top) and subsequently with a glyceraldehyde 3-phosphate dehydrogenase (Gapdh) cDNA probe which serves as a loading control (bottom). () Photograph of an agarose gel containing PCR amplimers derived from genomic DNA of mice with different Lrp5 genotypes. () Graphs depicting the areal bone mi! neral density (aBMD) measured by dual-energy X-ray absorptiometry (DEXA) in mice with different Lrp5 genotypes; mice followed until the age of 16.5 weeks (top and middle). Graphs depicting the percentage of trabecular bone volume in the total volume (BV/TV) of the distal femora and fifth lumbar vertebrae of 16.5-week-old male and female mice with different Lrp5 genotypes (bottom). () Representative μCT scan images obtained from 16.5-week-old mice with different Lrp5 genotypes. Scale bars, 1 mm. () Graphs depicting biomechanical properties of whole femora in a three-point bending assay from 16.5-week-old mice with different Lrp5 genotypes. () Representative images of new bone formation assessed by double calcein labeling of mice with different Lrp5 genotypes. Bone formation rates/bone surface area (BFR/BS); mineral apposition rates (MAR); mineralizing surface/bone surface (MS/BS). The numbers of mice studied are indicated. Error bars show means ± s.d. *P < 0.05 versus WT m! ice; #P < 0.05 versus heterozygous Lrp5 HBM mice. * Figure 2: Effect of activating Lrp5 NeoR-containing HBM alleles. () Graphs depicting femoral and vertebral trabecular BV/TV in WT mice and in mice with Lrp5 NeoR-containing HBM alleles. (,) Graphs depicting femoral trabecular BV/TV () and vertebral trabecular BV/TV () in mice with (shaded bars) and without (unshaded bars) inherited NeoR-containing Lrp5 HBM alleles (+/AN or +/GN, and +/A or +/G, respectively), and with and without Vil1-Cre (+V and –V, respectively) or Dmp1-Cre (+D and –D, respectively) transgenes. () Photographs of agarose gels containing PCR amplimers derived from mouse genomic DNA extracted from either duodenum or femur cortex of mice with different Lrp5 and Cre-transgene genotypes. PCR amplimers correspond to the sizes depicted in Figure 1a. Top, amplimers from the Dmp1-Cre cross. Bottom, amplimers from the Vil1-Cre cross. WT allele (arrowheads), AN or GN allele (double arrowheads) A or G allele (arrows). () Graphs depicting fluorochrome-derived bone formation parameters in the distal femur from 9-week-old female mi! ce that were administered double calcein labeling. Group notations (x axis) follow those described for panel . () Graphs depicting the proportion of distal femur trabecular bone surface covered by osteoclasts (Oc.S; left) and osteoblasts (Ob.S; right). () Graphs depicting femoral and vertebral trabecular BV/TV in 12-week-old mice with (shaded bars) and without (unshaded bars) inherited NeoR-containing Lrp5 HBM alleles (+/AN and +/A), respectively, and with and without the Prrx1-Cre transgene (+P and −P, respectively). MS/BS, MAR, BFR/BS are as defined in Figure 1. The numbers of mice studied are indicated. Error bars show means ± s.d. *P < 0.05 versus NeoR-containing littermates that did not inherit a Cre transgene. * Figure 3: Generation and characterization of mice with a conditional knockout allele of Lrp5. () Schematic depicting the creation of the Lrp5 floxed allele. loxP sites (arrowheads), the neomycin-resistance cassette (NeoR), flippase (Flp) recognition target (FRT) sites (diamonds) and the diphtheria toxin (DT) cassette are shown. The relative locations and orientation of the three primers (arrows) used for PCR genotyping and their expected amplimer sizes are noted. () Photograph of agarose gel depicting PCR amplimers for WT (+), floxed (f) and knockout (−) Lrp5 alleles from genomic DNA of mice with different Lrp5 genotypes. () Photographs of agarose gels containing PCR amplimers derived from mouse genomic DNA extracted from either duodenum or femur cortex of floxed Lrp5 mice, with or without the Dmp1-Cre transgene (top) and with or without the Vil1-Cre transgene (bottom). () Graphs depicting whole-body aBMD (left), femoral trabecular BV/TV (middle) and vertebral trabecular BV/TV (right) in 16-week-old mice homozygous for WT or floxed Lrp5 alleles, with or without the! Dmp1-Cre transgene (+D and –D, respectively). () Graphs depicting whole-body aBMD and tibial trabecular BV/TV in 3-month-old and 12-month-old mice heterozygous or homozygous for floxed Lrp5 alleles with or without the Vil1-Cre transgene (+V and –V, respectively). The numbers of mice studied are indicated. Error bars show means ± s.d. *P < 0.05 versus floxed Lrp5 littermates that did not inherit the Cre transgene. * Figure 4: Effect of Lrp5 genotype on 5HT concentration and on Tph1 expression. () Graphs depicting whole-blood 5HT measured by HPLC in 6-month-old Lrp5 WT and knockout mice that had been backcrossed to C57BL6/J mice. Shown are 5HT measurements in Lrp5 WT and HBM-causing knock-in (G/G) mice on a mixed 129Sv/C57BL/6J background (far left); in 3-month-old Lrp5 WT, knockout and HBM knock-in (+/A) mice on a mixed 129Sv/C57BL/6J genetic background (middle left); and in 3-month-old male (middle right) and 13-month-old female (far right) WT and knockout littermates on a mixed 129SvEvBrd/ C57BL/6J-Tyrc-Brd background. () Graphs depicting the quantity of 5HT extracted from several regions of the intestine, beginning in the duodenum and proceeding through the jejunum, ileum and proximal colon in 3-month-old male (left) and in 13-month-old female (right) Lrp5 WT and knockout littermates on a mixed 129SvEvBrd/ C57BL/6J-Tyrc-Brd background. () Scattergram depicting vertebral trabecular BV/TV and whole blood 5HT measurements in individual Lrp5 WT (open symbols) and k! nockout (filled symbols) littermates. Correlations between BV/TV and whole blood serotonin were r2 = 0.13 (P = 0.16) for male mice, and r2 = 0.02 (P = 0.53) for female mice. () Graphs depicting normalized Tph1 transcript levels in duodenum RNA extracts from Lrp5 WT, knockout, and HBM-causing knock-in (+/A) mice on a mixed 129Sv/C57BL/6J genetic background (left) and duodenum and colon RNA extracts from Lrp5 WT and knockout mice on a 129SvEvBrd/ C57BL/6J-Tyrc-Brd background (right) with Gapdh serving as the internal control. The mean Tph1 expression level for Lrp5 WT duodenum is set as 100%. The numbers of mice studied are indicated. Error bars show means ± s.d. *P < 0.05 versus WT mice. * Figure 5: Bone mass in WT and Tph1−/− mice. () Graphs depicting femoral trabecular BV/TV in WT and Tph1−/− mice on either FVB/N or C57BL/6 backgrounds (left) or on a mixed 129SvEvBrd/C57BL/6J-Tyrc-Brd background (right). () Graphs depicting the vertebral trabecular BV/TV of the fifth lumbar vertebrae in the same mice described in panel . () Graphs depicting lumbar spine aBMD, as measured by DEXA, in the same mice described in panel . The numbers of mice studied are indicated. Error bars show means ± s.d. *P < 0.05 versus WT mice using an unpaired t test; none of these differences remain significant after correcting for multiple testing. * Figure 6: Bone mass after pharmacologic inhibition of Tph1 activity. () Graph depicting dose-dependent changes in 5HT content, compared to vehicle-treated controls, in 9-week-old WT female C57BL/6 mice after receiving daily doses of LP-923941 for 7 days. A daily dose of 250 mg kg−1 lowered 5HT content in whole blood and in intestine, but not in brain. () Graph depicting changes in the intestinal 5HT content in sham-operated (SHM) and ovariectomized (OVX) mice that received vehicle or LP-923941 (250 mg per kg per day) for 6 weeks. Ovariectomy alone reduced 5HT content in the duodenum and colon by ~12% (P < 0.05) compared with SHM mice. Treatment with LP-923941 significantly reduced serotonin content equally in all regions of the intestine in SHM and in OVX mice. () Effect of treatment with LP-923941 (250 mg per kg per day) or teriparatide (80 μg per kg per day), which is the 1–34 residue amino-terminal fragment of human parathyroid hormone (PTH), on serum P1NP levels (left), vertebral trabecular BV/TV (middle left), midshaft femur cortica! l thickness (Ct.th; middle right) and midshaft femoral volumetric BMD (right) in SHM and in OVX mice. () Effect of treating SHM and OVX rats with LP-923941 (50 or 250 mg per kg per day) or teriparatide (80 μg per kg per day) for 6 weeks on jejunal 5HT content (left), femoral trabecular BV/TV (middle) and vertebral trabecular BV/TV (right). The numbers of mice studied are indicated. Error bars show means ± s.d. *P < 0.05 versus vehicle-treated mice. Article preview Read the full article * Instant access to this article: US$32 Buy now * Subscribe to Nature Medicine for full access: Subscribe * Personal subscribers: Log in Additional access options: * Login via Athens * Login via your Institution * Purchase a site license * Use a document delivery service * Rent this article from DeepDyve * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. Author information * Abstract * Author information * Supplementary information Affiliations * Orthopedic Research Laboratories, Department of Orthopedic Surgery, Children's Hospital, Boston, Massachusetts, USA. * Yajun Cui, * Christina M Jacobsen & * Matthew L Warman * Department of Genetics, Case Western Reserve University School of Medicine, Cleveland, Ohio, USA. * Yajun Cui & * Ronald A Conlon * Weldon School of Biomedical Engineering, Purdue University, West Lafayette, Indiana, USA. * Paul J Niziolek * Department of Anatomy & Cell Biology, Indiana University School of Medicine, Indianapolis, Indiana, USA. * Paul J Niziolek & * Alexander G Robling * Department of Biomedical Engineering, Indiana University School of Medicine, Indianapolis, Indiana, USA. * Paul J Niziolek & * Alexander G Robling * F.M. Kirby Neurobiology Center, Children's Hospital, Boston, Massachusetts, USA. * Bryan T MacDonald & * Xi He * Department of Neurology, Harvard Medical School, Boston, Massachusetts, USA. * Bryan T MacDonald & * Xi He * Center for Skeletal Disease Research, Van Andel Research Institute, Grand Rapids, Michigan, USA. * Cassandra R Zylstra, * Daniel R Robinson, * Zhendong Zhong & * Bart O Williams * Max Delbrück Center for Molecular Medicine, Berlin-Buch, Germany. * Natalia Alenina, * Susann Matthes & * Michael Bader * Lexicon Pharmaceuticals, The Woodlands, Texas, USA. * Robert Brommage, * Qingyun Liu, * Faika Mseeh, * David R Powell, * Qi M Yang & * Brian Zambrowicz * Merck Sharp & Dohme Research Laboratories, Oss, The Netherlands. * Han Gerrits & * Jan A Gossen * Howard Hughes Medical Institute, Children's Hospital, Boston, Massachusetts, USA. * Matthew L Warman * Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA. * Matthew L Warman Contributions Y.C. created and did studies on the mice with the Lrp5 HBM alleles and measured serum serotonin levels by competitive ELISA. P.J.N. did radiographic imaging and biomechanical testing on the mice with HBM-associated alleles. B.T.M. contributed to the serotonin and Tph1 qRT-PCR measurements in HBM-causing and Lrp5 knockout mice. C.R.Z. did multiple studies using the conditional Lrp5 knockout mice. N.A. studied the Tph1−/− mice, and with S.M. measured whole blood serotonin levels from HBM-causing and Lrp5-knockout mice by HPLC. D.R.R. generated the conditional Lrp5 knockout strain and Z.Z. participated in conditional inactivation of this allele using different Cre transgenes. C.M.J. carried out the Prrx1-Cre experiments. R.B., F.M. and Q.M.Y. organized studies on Lrp5- and Tph -knockout mice, and also organized the mouse pharmacology experiment. H.G. and J.A.G. organized the rat pharmacology experiment. R.A.C., X.H., M.B., D.R.P., Q.L., B.Z., B.O.W., A.G.R. and M.L.W. desig! ned experiments and provided reagents and financial support. M.L.W. prepared the first draft of the manuscript. All co-authors contributed detailed methods and results, and revised and approved the manuscript. Competing financial interests Employees of Lexicon Pharmaceuticals (R.B., Q.L., F.M., D.R.P., Q.M.Y. and B.Z.) and Merck Sharp & Dohme Research Laboratories (H.G. and J.A.G.) have received compensation in the form of salary and stock options. Corresponding author Correspondence to: * Matthew L Warman Author Details * Yajun Cui Search for this author in: * NPG journals * PubMed * Google Scholar * Paul J Niziolek Search for this author in: * NPG journals * PubMed * Google Scholar * Bryan T MacDonald Search for this author in: * NPG journals * PubMed * Google Scholar * Cassandra R Zylstra Search for this author in: * NPG journals * PubMed * Google Scholar * Natalia Alenina Search for this author in: * NPG journals * PubMed * Google Scholar * Daniel R Robinson Search for this author in: * NPG journals * PubMed * Google Scholar * Zhendong Zhong Search for this author in: * NPG journals * PubMed * Google Scholar * Susann Matthes Search for this author in: * NPG journals * PubMed * Google Scholar * Christina M Jacobsen Search for this author in: * NPG journals * PubMed * Google Scholar * Ronald A Conlon Search for this author in: * NPG journals * PubMed * Google Scholar * Robert Brommage Search for this author in: * NPG journals * PubMed * Google Scholar * Qingyun Liu Search for this author in: * NPG journals * PubMed * Google Scholar * Faika Mseeh Search for this author in: * NPG journals * PubMed * Google Scholar * David R Powell Search for this author in: * NPG journals * PubMed * Google Scholar * Qi M Yang Search for this author in: * NPG journals * PubMed * Google Scholar * Brian Zambrowicz Search for this author in: * NPG journals * PubMed * Google Scholar * Han Gerrits Search for this author in: * NPG journals * PubMed * Google Scholar * Jan A Gossen Search for this author in: * NPG journals * PubMed * Google Scholar * Xi He Search for this author in: * NPG journals * PubMed * Google Scholar * Michael Bader Search for this author in: * NPG journals * PubMed * Google Scholar * Bart O Williams Search for this author in: * NPG journals * PubMed * Google Scholar * Matthew L Warman Contact Matthew L Warman Search for this author in: * NPG journals * PubMed * Google Scholar * Alexander G Robling Search for this author in: * NPG journals * PubMed * Google Scholar Supplementary information * Abstract * Author information * Supplementary information PDF files * Supplementary Text and Figures (9M) Supplementary Figures 1–10, Supplementary Tables 1–4 and Supplementary Methods Additional data
• ERasing breast cancer resistance through the kinome
  - Nat Med 17(6):660-661 (2011)
  Article preview View full access options Nature Medicine | News Market overlap points to irresponsible use of tuberculosis drugs * Erika Check-HaydenJournal name:Nature MedicineVolume: 17,Page:635Year published:(2011)DOI:doi:10.1038/nm0611-635Published online06 June 2011 sy5/sy5/ZUMA Press/Newscom The private drug market can treat two thirds of those who develop TB. People with active tuberculosis infections turn to the private market for treatment far more often than anyone had realized. And when they do, they encounter a chaotic array of treatment choices, many of which do not meet guidelines drawn up by the World Health Organization. These are the conclusions of a paper published on 4 May that counters the prevailing wisdom that the vast majority of people with tuberculosis are treated through publicly funded programs. Article preview Read the full article * Instant access to this article: US$32 Buy now * Subscribe to Nature Medicine for full access: Subscribe * Personal subscribers: Log in Additional access options: * Login via Athens * Login via your Institution * Purchase a site license * Use a document delivery service * Rent this article from DeepDyve * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. Additional data Author Details * Erika Check-Hayden Search for this author in: * NPG journals * PubMed * Google Scholar
• Imaging transplant rejection: a new view
  - Nat Med 17(6):662-663 (2011)
  Article preview View full access options Nature Medicine | Article Lrp5 functions in bone to regulate bone mass * Yajun Cui1, 2 * Paul J Niziolek3, 4, 5 * Bryan T MacDonald6, 7 * Cassandra R Zylstra8 * Natalia Alenina9 * Daniel R Robinson8 * Zhendong Zhong8 * Susann Matthes9 * Christina M Jacobsen1 * Ronald A Conlon2 * Robert Brommage10 * Qingyun Liu10 * Faika Mseeh10 * David R Powell10 * Qi M Yang10 * Brian Zambrowicz10 * Han Gerrits11 * Jan A Gossen11 * Xi He6, 7 * Michael Bader9 * Bart O Williams8 * Matthew L Warman1, 12, 13 * Alexander G Robling4, 5 * Affiliations * Contributions * Corresponding authorJournal name:Nature MedicineVolume: 17,Pages:684–691Year published:(2011)DOI:doi:10.1038/nm.2388Received14 December 2010Accepted27 April 2011Published online22 May 2011 Abstract * Abstract * Author information * Supplementary information Article tools * Full text * 日本語要約 * Print * Email * Download citation * Order reprints * Rights and permissions * Share/bookmark * Connotea * CiteULike * Facebook * Twitter * Delicious * Digg The human skeleton is affected by mutations in low-density lipoprotein receptor-related protein 5 (LRP5). To understand how LRP5 influences bone properties, we generated mice with osteocyte-specific expression of inducible Lrp5 mutations that cause high and low bone mass phenotypes in humans. We found that bone properties in these mice were comparable to bone properties in mice with inherited mutations. We also induced an Lrp5 mutation in cells that form the appendicular skeleton but not in cells that form the axial skeleton; we observed that bone properties were altered in the limb but not in the spine. These data indicate that Lrp5 signaling functions locally, and they suggest that increasing LRP5 signaling in mature bone cells may be a strategy for treating human disorders associated with low bone mass, such as osteoporosis. Figures at a glance * Figure 1: Generation and characterization of HBM-causing Lrp5 knock-in mice. () Schematic depicting the targeting and genotyping strategies for Lrp5 HBM alleles. The 5′ targeting arm contains a neomycin-resistance cassette (NeoR) flanked by loxP sites (arrowheads). The 3′ targeting arm begins in intron 2 and extends into intron 4, and is followed by a thymidine kinase (TK) cassette. Site-directed mutagenesis altered specific amino acid residues encoded by Lrp5 exon 3 (asterisks). The relative locations and orientation of primers used for PCR genotyping and their expected amplimer sizes are noted. () Autoradiographs of a northern blot containing whole-bone total RNA from mice with different Lrp5 genotypes initially hybridized with a radioactive Lrp5 cDNA probe (top) and subsequently with a glyceraldehyde 3-phosphate dehydrogenase (Gapdh) cDNA probe which serves as a loading control (bottom). () Photograph of an agarose gel containing PCR amplimers derived from genomic DNA of mice with different Lrp5 genotypes. () Graphs depicting the areal bone mi! neral density (aBMD) measured by dual-energy X-ray absorptiometry (DEXA) in mice with different Lrp5 genotypes; mice followed until the age of 16.5 weeks (top and middle). Graphs depicting the percentage of trabecular bone volume in the total volume (BV/TV) of the distal femora and fifth lumbar vertebrae of 16.5-week-old male and female mice with different Lrp5 genotypes (bottom). () Representative μCT scan images obtained from 16.5-week-old mice with different Lrp5 genotypes. Scale bars, 1 mm. () Graphs depicting biomechanical properties of whole femora in a three-point bending assay from 16.5-week-old mice with different Lrp5 genotypes. () Representative images of new bone formation assessed by double calcein labeling of mice with different Lrp5 genotypes. Bone formation rates/bone surface area (BFR/BS); mineral apposition rates (MAR); mineralizing surface/bone surface (MS/BS). The numbers of mice studied are indicated. Error bars show means ± s.d. *P < 0.05 versus WT m! ice; #P < 0.05 versus heterozygous Lrp5 HBM mice. * Figure 2: Effect of activating Lrp5 NeoR-containing HBM alleles. () Graphs depicting femoral and vertebral trabecular BV/TV in WT mice and in mice with Lrp5 NeoR-containing HBM alleles. (,) Graphs depicting femoral trabecular BV/TV () and vertebral trabecular BV/TV () in mice with (shaded bars) and without (unshaded bars) inherited NeoR-containing Lrp5 HBM alleles (+/AN or +/GN, and +/A or +/G, respectively), and with and without Vil1-Cre (+V and –V, respectively) or Dmp1-Cre (+D and –D, respectively) transgenes. () Photographs of agarose gels containing PCR amplimers derived from mouse genomic DNA extracted from either duodenum or femur cortex of mice with different Lrp5 and Cre-transgene genotypes. PCR amplimers correspond to the sizes depicted in Figure 1a. Top, amplimers from the Dmp1-Cre cross. Bottom, amplimers from the Vil1-Cre cross. WT allele (arrowheads), AN or GN allele (double arrowheads) A or G allele (arrows). () Graphs depicting fluorochrome-derived bone formation parameters in the distal femur from 9-week-old female mi! ce that were administered double calcein labeling. Group notations (x axis) follow those described for panel . () Graphs depicting the proportion of distal femur trabecular bone surface covered by osteoclasts (Oc.S; left) and osteoblasts (Ob.S; right). () Graphs depicting femoral and vertebral trabecular BV/TV in 12-week-old mice with (shaded bars) and without (unshaded bars) inherited NeoR-containing Lrp5 HBM alleles (+/AN and +/A), respectively, and with and without the Prrx1-Cre transgene (+P and −P, respectively). MS/BS, MAR, BFR/BS are as defined in Figure 1. The numbers of mice studied are indicated. Error bars show means ± s.d. *P < 0.05 versus NeoR-containing littermates that did not inherit a Cre transgene. * Figure 3: Generation and characterization of mice with a conditional knockout allele of Lrp5. () Schematic depicting the creation of the Lrp5 floxed allele. loxP sites (arrowheads), the neomycin-resistance cassette (NeoR), flippase (Flp) recognition target (FRT) sites (diamonds) and the diphtheria toxin (DT) cassette are shown. The relative locations and orientation of the three primers (arrows) used for PCR genotyping and their expected amplimer sizes are noted. () Photograph of agarose gel depicting PCR amplimers for WT (+), floxed (f) and knockout (−) Lrp5 alleles from genomic DNA of mice with different Lrp5 genotypes. () Photographs of agarose gels containing PCR amplimers derived from mouse genomic DNA extracted from either duodenum or femur cortex of floxed Lrp5 mice, with or without the Dmp1-Cre transgene (top) and with or without the Vil1-Cre transgene (bottom). () Graphs depicting whole-body aBMD (left), femoral trabecular BV/TV (middle) and vertebral trabecular BV/TV (right) in 16-week-old mice homozygous for WT or floxed Lrp5 alleles, with or without the! Dmp1-Cre transgene (+D and –D, respectively). () Graphs depicting whole-body aBMD and tibial trabecular BV/TV in 3-month-old and 12-month-old mice heterozygous or homozygous for floxed Lrp5 alleles with or without the Vil1-Cre transgene (+V and –V, respectively). The numbers of mice studied are indicated. Error bars show means ± s.d. *P < 0.05 versus floxed Lrp5 littermates that did not inherit the Cre transgene. * Figure 4: Effect of Lrp5 genotype on 5HT concentration and on Tph1 expression. () Graphs depicting whole-blood 5HT measured by HPLC in 6-month-old Lrp5 WT and knockout mice that had been backcrossed to C57BL6/J mice. Shown are 5HT measurements in Lrp5 WT and HBM-causing knock-in (G/G) mice on a mixed 129Sv/C57BL/6J background (far left); in 3-month-old Lrp5 WT, knockout and HBM knock-in (+/A) mice on a mixed 129Sv/C57BL/6J genetic background (middle left); and in 3-month-old male (middle right) and 13-month-old female (far right) WT and knockout littermates on a mixed 129SvEvBrd/ C57BL/6J-Tyrc-Brd background. () Graphs depicting the quantity of 5HT extracted from several regions of the intestine, beginning in the duodenum and proceeding through the jejunum, ileum and proximal colon in 3-month-old male (left) and in 13-month-old female (right) Lrp5 WT and knockout littermates on a mixed 129SvEvBrd/ C57BL/6J-Tyrc-Brd background. () Scattergram depicting vertebral trabecular BV/TV and whole blood 5HT measurements in individual Lrp5 WT (open symbols) and k! nockout (filled symbols) littermates. Correlations between BV/TV and whole blood serotonin were r2 = 0.13 (P = 0.16) for male mice, and r2 = 0.02 (P = 0.53) for female mice. () Graphs depicting normalized Tph1 transcript levels in duodenum RNA extracts from Lrp5 WT, knockout, and HBM-causing knock-in (+/A) mice on a mixed 129Sv/C57BL/6J genetic background (left) and duodenum and colon RNA extracts from Lrp5 WT and knockout mice on a 129SvEvBrd/ C57BL/6J-Tyrc-Brd background (right) with Gapdh serving as the internal control. The mean Tph1 expression level for Lrp5 WT duodenum is set as 100%. The numbers of mice studied are indicated. Error bars show means ± s.d. *P < 0.05 versus WT mice. * Figure 5: Bone mass in WT and Tph1−/− mice. () Graphs depicting femoral trabecular BV/TV in WT and Tph1−/− mice on either FVB/N or C57BL/6 backgrounds (left) or on a mixed 129SvEvBrd/C57BL/6J-Tyrc-Brd background (right). () Graphs depicting the vertebral trabecular BV/TV of the fifth lumbar vertebrae in the same mice described in panel . () Graphs depicting lumbar spine aBMD, as measured by DEXA, in the same mice described in panel . The numbers of mice studied are indicated. Error bars show means ± s.d. *P < 0.05 versus WT mice using an unpaired t test; none of these differences remain significant after correcting for multiple testing. * Figure 6: Bone mass after pharmacologic inhibition of Tph1 activity. () Graph depicting dose-dependent changes in 5HT content, compared to vehicle-treated controls, in 9-week-old WT female C57BL/6 mice after receiving daily doses of LP-923941 for 7 days. A daily dose of 250 mg kg−1 lowered 5HT content in whole blood and in intestine, but not in brain. () Graph depicting changes in the intestinal 5HT content in sham-operated (SHM) and ovariectomized (OVX) mice that received vehicle or LP-923941 (250 mg per kg per day) for 6 weeks. Ovariectomy alone reduced 5HT content in the duodenum and colon by ~12% (P < 0.05) compared with SHM mice. Treatment with LP-923941 significantly reduced serotonin content equally in all regions of the intestine in SHM and in OVX mice. () Effect of treatment with LP-923941 (250 mg per kg per day) or teriparatide (80 μg per kg per day), which is the 1–34 residue amino-terminal fragment of human parathyroid hormone (PTH), on serum P1NP levels (left), vertebral trabecular BV/TV (middle left), midshaft femur cortica! l thickness (Ct.th; middle right) and midshaft femoral volumetric BMD (right) in SHM and in OVX mice. () Effect of treating SHM and OVX rats with LP-923941 (50 or 250 mg per kg per day) or teriparatide (80 μg per kg per day) for 6 weeks on jejunal 5HT content (left), femoral trabecular BV/TV (middle) and vertebral trabecular BV/TV (right). The numbers of mice studied are indicated. Error bars show means ± s.d. *P < 0.05 versus vehicle-treated mice. Article preview Read the full article * Instant access to this article: US$32 Buy now * Subscribe to Nature Medicine for full access: Subscribe * Personal subscribers: Log in Additional access options: * Login via Athens * Login via your Institution * Purchase a site license * Use a document delivery service * Rent this article from DeepDyve * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. Author information * Abstract * Author information * Supplementary information Affiliations * Orthopedic Research Laboratories, Department of Orthopedic Surgery, Children's Hospital, Boston, Massachusetts, USA. * Yajun Cui, * Christina M Jacobsen & * Matthew L Warman * Department of Genetics, Case Western Reserve University School of Medicine, Cleveland, Ohio, USA. * Yajun Cui & * Ronald A Conlon * Weldon School of Biomedical Engineering, Purdue University, West Lafayette, Indiana, USA. * Paul J Niziolek * Department of Anatomy & Cell Biology, Indiana University School of Medicine, Indianapolis, Indiana, USA. * Paul J Niziolek & * Alexander G Robling * Department of Biomedical Engineering, Indiana University School of Medicine, Indianapolis, Indiana, USA. * Paul J Niziolek & * Alexander G Robling * F.M. Kirby Neurobiology Center, Children's Hospital, Boston, Massachusetts, USA. * Bryan T MacDonald & * Xi He * Department of Neurology, Harvard Medical School, Boston, Massachusetts, USA. * Bryan T MacDonald & * Xi He * Center for Skeletal Disease Research, Van Andel Research Institute, Grand Rapids, Michigan, USA. * Cassandra R Zylstra, * Daniel R Robinson, * Zhendong Zhong & * Bart O Williams * Max Delbrück Center for Molecular Medicine, Berlin-Buch, Germany. * Natalia Alenina, * Susann Matthes & * Michael Bader * Lexicon Pharmaceuticals, The Woodlands, Texas, USA. * Robert Brommage, * Qingyun Liu, * Faika Mseeh, * David R Powell, * Qi M Yang & * Brian Zambrowicz * Merck Sharp & Dohme Research Laboratories, Oss, The Netherlands. * Han Gerrits & * Jan A Gossen * Howard Hughes Medical Institute, Children's Hospital, Boston, Massachusetts, USA. * Matthew L Warman * Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA. * Matthew L Warman Contributions Y.C. created and did studies on the mice with the Lrp5 HBM alleles and measured serum serotonin levels by competitive ELISA. P.J.N. did radiographic imaging and biomechanical testing on the mice with HBM-associated alleles. B.T.M. contributed to the serotonin and Tph1 qRT-PCR measurements in HBM-causing and Lrp5 knockout mice. C.R.Z. did multiple studies using the conditional Lrp5 knockout mice. N.A. studied the Tph1−/− mice, and with S.M. measured whole blood serotonin levels from HBM-causing and Lrp5-knockout mice by HPLC. D.R.R. generated the conditional Lrp5 knockout strain and Z.Z. participated in conditional inactivation of this allele using different Cre transgenes. C.M.J. carried out the Prrx1-Cre experiments. R.B., F.M. and Q.M.Y. organized studies on Lrp5- and Tph -knockout mice, and also organized the mouse pharmacology experiment. H.G. and J.A.G. organized the rat pharmacology experiment. R.A.C., X.H., M.B., D.R.P., Q.L., B.Z., B.O.W., A.G.R. and M.L.W. desig! ned experiments and provided reagents and financial support. M.L.W. prepared the first draft of the manuscript. All co-authors contributed detailed methods and results, and revised and approved the manuscript. Competing financial interests Employees of Lexicon Pharmaceuticals (R.B., Q.L., F.M., D.R.P., Q.M.Y. and B.Z.) and Merck Sharp & Dohme Research Laboratories (H.G. and J.A.G.) have received compensation in the form of salary and stock options. Corresponding author Correspondence to: * Matthew L Warman Author Details * Yajun Cui Search for this author in: * NPG journals * PubMed * Google Scholar * Paul J Niziolek Search for this author in: * NPG journals * PubMed * Google Scholar * Bryan T MacDonald Search for this author in: * NPG journals * PubMed * Google Scholar * Cassandra R Zylstra Search for this author in: * NPG journals * PubMed * Google Scholar * Natalia Alenina Search for this author in: * NPG journals * PubMed * Google Scholar * Daniel R Robinson Search for this author in: * NPG journals * PubMed * Google Scholar * Zhendong Zhong Search for this author in: * NPG journals * PubMed * Google Scholar * Susann Matthes Search for this author in: * NPG journals * PubMed * Google Scholar * Christina M Jacobsen Search for this author in: * NPG journals * PubMed * Google Scholar * Ronald A Conlon Search for this author in: * NPG journals * PubMed * Google Scholar * Robert Brommage Search for this author in: * NPG journals * PubMed * Google Scholar * Qingyun Liu Search for this author in: * NPG journals * PubMed * Google Scholar * Faika Mseeh Search for this author in: * NPG journals * PubMed * Google Scholar * David R Powell Search for this author in: * NPG journals * PubMed * Google Scholar * Qi M Yang Search for this author in: * NPG journals * PubMed * Google Scholar * Brian Zambrowicz Search for this author in: * NPG journals * PubMed * Google Scholar * Han Gerrits Search for this author in: * NPG journals * PubMed * Google Scholar * Jan A Gossen Search for this author in: * NPG journals * PubMed * Google Scholar * Xi He Search for this author in: * NPG journals * PubMed * Google Scholar * Michael Bader Search for this author in: * NPG journals * PubMed * Google Scholar * Bart O Williams Search for this author in: * NPG journals * PubMed * Google Scholar * Matthew L Warman Contact Matthew L Warman Search for this author in: * NPG journals * PubMed * Google Scholar * Alexander G Robling Search for this author in: * NPG journals * PubMed * Google Scholar Supplementary information * Abstract * Author information * Supplementary information PDF files * Supplementary Text and Figures (9M) Supplementary Figures 1–10, Supplementary Tables 1–4 and Supplementary Methods Additional data
• Wormless without wingless
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  Article preview View full access options Nature Medicine | Article Lrp5 functions in bone to regulate bone mass * Yajun Cui1, 2 * Paul J Niziolek3, 4, 5 * Bryan T MacDonald6, 7 * Cassandra R Zylstra8 * Natalia Alenina9 * Daniel R Robinson8 * Zhendong Zhong8 * Susann Matthes9 * Christina M Jacobsen1 * Ronald A Conlon2 * Robert Brommage10 * Qingyun Liu10 * Faika Mseeh10 * David R Powell10 * Qi M Yang10 * Brian Zambrowicz10 * Han Gerrits11 * Jan A Gossen11 * Xi He6, 7 * Michael Bader9 * Bart O Williams8 * Matthew L Warman1, 12, 13 * Alexander G Robling4, 5 * Affiliations * Contributions * Corresponding authorJournal name:Nature MedicineVolume: 17,Pages:684–691Year published:(2011)DOI:doi:10.1038/nm.2388Received14 December 2010Accepted27 April 2011Published online22 May 2011 Abstract * Abstract * Author information * Supplementary information Article tools * Full text * 日本語要約 * Print * Email * Download citation * Order reprints * Rights and permissions * Share/bookmark * Connotea * CiteULike * Facebook * Twitter * Delicious * Digg The human skeleton is affected by mutations in low-density lipoprotein receptor-related protein 5 (LRP5). To understand how LRP5 influences bone properties, we generated mice with osteocyte-specific expression of inducible Lrp5 mutations that cause high and low bone mass phenotypes in humans. We found that bone properties in these mice were comparable to bone properties in mice with inherited mutations. We also induced an Lrp5 mutation in cells that form the appendicular skeleton but not in cells that form the axial skeleton; we observed that bone properties were altered in the limb but not in the spine. These data indicate that Lrp5 signaling functions locally, and they suggest that increasing LRP5 signaling in mature bone cells may be a strategy for treating human disorders associated with low bone mass, such as osteoporosis. Figures at a glance * Figure 1: Generation and characterization of HBM-causing Lrp5 knock-in mice. () Schematic depicting the targeting and genotyping strategies for Lrp5 HBM alleles. The 5′ targeting arm contains a neomycin-resistance cassette (NeoR) flanked by loxP sites (arrowheads). The 3′ targeting arm begins in intron 2 and extends into intron 4, and is followed by a thymidine kinase (TK) cassette. Site-directed mutagenesis altered specific amino acid residues encoded by Lrp5 exon 3 (asterisks). The relative locations and orientation of primers used for PCR genotyping and their expected amplimer sizes are noted. () Autoradiographs of a northern blot containing whole-bone total RNA from mice with different Lrp5 genotypes initially hybridized with a radioactive Lrp5 cDNA probe (top) and subsequently with a glyceraldehyde 3-phosphate dehydrogenase (Gapdh) cDNA probe which serves as a loading control (bottom). () Photograph of an agarose gel containing PCR amplimers derived from genomic DNA of mice with different Lrp5 genotypes. () Graphs depicting the areal bone mi! neral density (aBMD) measured by dual-energy X-ray absorptiometry (DEXA) in mice with different Lrp5 genotypes; mice followed until the age of 16.5 weeks (top and middle). Graphs depicting the percentage of trabecular bone volume in the total volume (BV/TV) of the distal femora and fifth lumbar vertebrae of 16.5-week-old male and female mice with different Lrp5 genotypes (bottom). () Representative μCT scan images obtained from 16.5-week-old mice with different Lrp5 genotypes. Scale bars, 1 mm. () Graphs depicting biomechanical properties of whole femora in a three-point bending assay from 16.5-week-old mice with different Lrp5 genotypes. () Representative images of new bone formation assessed by double calcein labeling of mice with different Lrp5 genotypes. Bone formation rates/bone surface area (BFR/BS); mineral apposition rates (MAR); mineralizing surface/bone surface (MS/BS). The numbers of mice studied are indicated. Error bars show means ± s.d. *P < 0.05 versus WT m! ice; #P < 0.05 versus heterozygous Lrp5 HBM mice. * Figure 2: Effect of activating Lrp5 NeoR-containing HBM alleles. () Graphs depicting femoral and vertebral trabecular BV/TV in WT mice and in mice with Lrp5 NeoR-containing HBM alleles. (,) Graphs depicting femoral trabecular BV/TV () and vertebral trabecular BV/TV () in mice with (shaded bars) and without (unshaded bars) inherited NeoR-containing Lrp5 HBM alleles (+/AN or +/GN, and +/A or +/G, respectively), and with and without Vil1-Cre (+V and –V, respectively) or Dmp1-Cre (+D and –D, respectively) transgenes. () Photographs of agarose gels containing PCR amplimers derived from mouse genomic DNA extracted from either duodenum or femur cortex of mice with different Lrp5 and Cre-transgene genotypes. PCR amplimers correspond to the sizes depicted in Figure 1a. Top, amplimers from the Dmp1-Cre cross. Bottom, amplimers from the Vil1-Cre cross. WT allele (arrowheads), AN or GN allele (double arrowheads) A or G allele (arrows). () Graphs depicting fluorochrome-derived bone formation parameters in the distal femur from 9-week-old female mi! ce that were administered double calcein labeling. Group notations (x axis) follow those described for panel . () Graphs depicting the proportion of distal femur trabecular bone surface covered by osteoclasts (Oc.S; left) and osteoblasts (Ob.S; right). () Graphs depicting femoral and vertebral trabecular BV/TV in 12-week-old mice with (shaded bars) and without (unshaded bars) inherited NeoR-containing Lrp5 HBM alleles (+/AN and +/A), respectively, and with and without the Prrx1-Cre transgene (+P and −P, respectively). MS/BS, MAR, BFR/BS are as defined in Figure 1. The numbers of mice studied are indicated. Error bars show means ± s.d. *P < 0.05 versus NeoR-containing littermates that did not inherit a Cre transgene. * Figure 3: Generation and characterization of mice with a conditional knockout allele of Lrp5. () Schematic depicting the creation of the Lrp5 floxed allele. loxP sites (arrowheads), the neomycin-resistance cassette (NeoR), flippase (Flp) recognition target (FRT) sites (diamonds) and the diphtheria toxin (DT) cassette are shown. The relative locations and orientation of the three primers (arrows) used for PCR genotyping and their expected amplimer sizes are noted. () Photograph of agarose gel depicting PCR amplimers for WT (+), floxed (f) and knockout (−) Lrp5 alleles from genomic DNA of mice with different Lrp5 genotypes. () Photographs of agarose gels containing PCR amplimers derived from mouse genomic DNA extracted from either duodenum or femur cortex of floxed Lrp5 mice, with or without the Dmp1-Cre transgene (top) and with or without the Vil1-Cre transgene (bottom). () Graphs depicting whole-body aBMD (left), femoral trabecular BV/TV (middle) and vertebral trabecular BV/TV (right) in 16-week-old mice homozygous for WT or floxed Lrp5 alleles, with or without the! Dmp1-Cre transgene (+D and –D, respectively). () Graphs depicting whole-body aBMD and tibial trabecular BV/TV in 3-month-old and 12-month-old mice heterozygous or homozygous for floxed Lrp5 alleles with or without the Vil1-Cre transgene (+V and –V, respectively). The numbers of mice studied are indicated. Error bars show means ± s.d. *P < 0.05 versus floxed Lrp5 littermates that did not inherit the Cre transgene. * Figure 4: Effect of Lrp5 genotype on 5HT concentration and on Tph1 expression. () Graphs depicting whole-blood 5HT measured by HPLC in 6-month-old Lrp5 WT and knockout mice that had been backcrossed to C57BL6/J mice. Shown are 5HT measurements in Lrp5 WT and HBM-causing knock-in (G/G) mice on a mixed 129Sv/C57BL/6J background (far left); in 3-month-old Lrp5 WT, knockout and HBM knock-in (+/A) mice on a mixed 129Sv/C57BL/6J genetic background (middle left); and in 3-month-old male (middle right) and 13-month-old female (far right) WT and knockout littermates on a mixed 129SvEvBrd/ C57BL/6J-Tyrc-Brd background. () Graphs depicting the quantity of 5HT extracted from several regions of the intestine, beginning in the duodenum and proceeding through the jejunum, ileum and proximal colon in 3-month-old male (left) and in 13-month-old female (right) Lrp5 WT and knockout littermates on a mixed 129SvEvBrd/ C57BL/6J-Tyrc-Brd background. () Scattergram depicting vertebral trabecular BV/TV and whole blood 5HT measurements in individual Lrp5 WT (open symbols) and k! nockout (filled symbols) littermates. Correlations between BV/TV and whole blood serotonin were r2 = 0.13 (P = 0.16) for male mice, and r2 = 0.02 (P = 0.53) for female mice. () Graphs depicting normalized Tph1 transcript levels in duodenum RNA extracts from Lrp5 WT, knockout, and HBM-causing knock-in (+/A) mice on a mixed 129Sv/C57BL/6J genetic background (left) and duodenum and colon RNA extracts from Lrp5 WT and knockout mice on a 129SvEvBrd/ C57BL/6J-Tyrc-Brd background (right) with Gapdh serving as the internal control. The mean Tph1 expression level for Lrp5 WT duodenum is set as 100%. The numbers of mice studied are indicated. Error bars show means ± s.d. *P < 0.05 versus WT mice. * Figure 5: Bone mass in WT and Tph1−/− mice. () Graphs depicting femoral trabecular BV/TV in WT and Tph1−/− mice on either FVB/N or C57BL/6 backgrounds (left) or on a mixed 129SvEvBrd/C57BL/6J-Tyrc-Brd background (right). () Graphs depicting the vertebral trabecular BV/TV of the fifth lumbar vertebrae in the same mice described in panel . () Graphs depicting lumbar spine aBMD, as measured by DEXA, in the same mice described in panel . The numbers of mice studied are indicated. Error bars show means ± s.d. *P < 0.05 versus WT mice using an unpaired t test; none of these differences remain significant after correcting for multiple testing. * Figure 6: Bone mass after pharmacologic inhibition of Tph1 activity. () Graph depicting dose-dependent changes in 5HT content, compared to vehicle-treated controls, in 9-week-old WT female C57BL/6 mice after receiving daily doses of LP-923941 for 7 days. A daily dose of 250 mg kg−1 lowered 5HT content in whole blood and in intestine, but not in brain. () Graph depicting changes in the intestinal 5HT content in sham-operated (SHM) and ovariectomized (OVX) mice that received vehicle or LP-923941 (250 mg per kg per day) for 6 weeks. Ovariectomy alone reduced 5HT content in the duodenum and colon by ~12% (P < 0.05) compared with SHM mice. Treatment with LP-923941 significantly reduced serotonin content equally in all regions of the intestine in SHM and in OVX mice. () Effect of treatment with LP-923941 (250 mg per kg per day) or teriparatide (80 μg per kg per day), which is the 1–34 residue amino-terminal fragment of human parathyroid hormone (PTH), on serum P1NP levels (left), vertebral trabecular BV/TV (middle left), midshaft femur cortica! l thickness (Ct.th; middle right) and midshaft femoral volumetric BMD (right) in SHM and in OVX mice. () Effect of treating SHM and OVX rats with LP-923941 (50 or 250 mg per kg per day) or teriparatide (80 μg per kg per day) for 6 weeks on jejunal 5HT content (left), femoral trabecular BV/TV (middle) and vertebral trabecular BV/TV (right). The numbers of mice studied are indicated. Error bars show means ± s.d. *P < 0.05 versus vehicle-treated mice. Article preview Read the full article * Instant access to this article: US$32 Buy now * Subscribe to Nature Medicine for full access: Subscribe * Personal subscribers: Log in Additional access options: * Login via Athens * Login via your Institution * Purchase a site license * Use a document delivery service * Rent this article from DeepDyve * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. Author information * Abstract * Author information * Supplementary information Affiliations * Orthopedic Research Laboratories, Department of Orthopedic Surgery, Children's Hospital, Boston, Massachusetts, USA. * Yajun Cui, * Christina M Jacobsen & * Matthew L Warman * Department of Genetics, Case Western Reserve University School of Medicine, Cleveland, Ohio, USA. * Yajun Cui & * Ronald A Conlon * Weldon School of Biomedical Engineering, Purdue University, West Lafayette, Indiana, USA. * Paul J Niziolek * Department of Anatomy & Cell Biology, Indiana University School of Medicine, Indianapolis, Indiana, USA. * Paul J Niziolek & * Alexander G Robling * Department of Biomedical Engineering, Indiana University School of Medicine, Indianapolis, Indiana, USA. * Paul J Niziolek & * Alexander G Robling * F.M. Kirby Neurobiology Center, Children's Hospital, Boston, Massachusetts, USA. * Bryan T MacDonald & * Xi He * Department of Neurology, Harvard Medical School, Boston, Massachusetts, USA. * Bryan T MacDonald & * Xi He * Center for Skeletal Disease Research, Van Andel Research Institute, Grand Rapids, Michigan, USA. * Cassandra R Zylstra, * Daniel R Robinson, * Zhendong Zhong & * Bart O Williams * Max Delbrück Center for Molecular Medicine, Berlin-Buch, Germany. * Natalia Alenina, * Susann Matthes & * Michael Bader * Lexicon Pharmaceuticals, The Woodlands, Texas, USA. * Robert Brommage, * Qingyun Liu, * Faika Mseeh, * David R Powell, * Qi M Yang & * Brian Zambrowicz * Merck Sharp & Dohme Research Laboratories, Oss, The Netherlands. * Han Gerrits & * Jan A Gossen * Howard Hughes Medical Institute, Children's Hospital, Boston, Massachusetts, USA. * Matthew L Warman * Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA. * Matthew L Warman Contributions Y.C. created and did studies on the mice with the Lrp5 HBM alleles and measured serum serotonin levels by competitive ELISA. P.J.N. did radiographic imaging and biomechanical testing on the mice with HBM-associated alleles. B.T.M. contributed to the serotonin and Tph1 qRT-PCR measurements in HBM-causing and Lrp5 knockout mice. C.R.Z. did multiple studies using the conditional Lrp5 knockout mice. N.A. studied the Tph1−/− mice, and with S.M. measured whole blood serotonin levels from HBM-causing and Lrp5-knockout mice by HPLC. D.R.R. generated the conditional Lrp5 knockout strain and Z.Z. participated in conditional inactivation of this allele using different Cre transgenes. C.M.J. carried out the Prrx1-Cre experiments. R.B., F.M. and Q.M.Y. organized studies on Lrp5- and Tph -knockout mice, and also organized the mouse pharmacology experiment. H.G. and J.A.G. organized the rat pharmacology experiment. R.A.C., X.H., M.B., D.R.P., Q.L., B.Z., B.O.W., A.G.R. and M.L.W. desig! ned experiments and provided reagents and financial support. M.L.W. prepared the first draft of the manuscript. All co-authors contributed detailed methods and results, and revised and approved the manuscript. Competing financial interests Employees of Lexicon Pharmaceuticals (R.B., Q.L., F.M., D.R.P., Q.M.Y. and B.Z.) and Merck Sharp & Dohme Research Laboratories (H.G. and J.A.G.) have received compensation in the form of salary and stock options. Corresponding author Correspondence to: * Matthew L Warman Author Details * Yajun Cui Search for this author in: * NPG journals * PubMed * Google Scholar * Paul J Niziolek Search for this author in: * NPG journals * PubMed * Google Scholar * Bryan T MacDonald Search for this author in: * NPG journals * PubMed * Google Scholar * Cassandra R Zylstra Search for this author in: * NPG journals * PubMed * Google Scholar * Natalia Alenina Search for this author in: * NPG journals * PubMed * Google Scholar * Daniel R Robinson Search for this author in: * NPG journals * PubMed * Google Scholar * Zhendong Zhong Search for this author in: * NPG journals * PubMed * Google Scholar * Susann Matthes Search for this author in: * NPG journals * PubMed * Google Scholar * Christina M Jacobsen Search for this author in: * NPG journals * PubMed * Google Scholar * Ronald A Conlon Search for this author in: * NPG journals * PubMed * Google Scholar * Robert Brommage Search for this author in: * NPG journals * PubMed * Google Scholar * Qingyun Liu Search for this author in: * NPG journals * PubMed * Google Scholar * Faika Mseeh Search for this author in: * NPG journals * PubMed * Google Scholar * David R Powell Search for this author in: * NPG journals * PubMed * Google Scholar * Qi M Yang Search for this author in: * NPG journals * PubMed * Google Scholar * Brian Zambrowicz Search for this author in: * NPG journals * PubMed * Google Scholar * Han Gerrits Search for this author in: * NPG journals * PubMed * Google Scholar * Jan A Gossen Search for this author in: * NPG journals * PubMed * Google Scholar * Xi He Search for this author in: * NPG journals * PubMed * Google Scholar * Michael Bader Search for this author in: * NPG journals * PubMed * Google Scholar * Bart O Williams Search for this author in: * NPG journals * PubMed * Google Scholar * Matthew L Warman Contact Matthew L Warman Search for this author in: * NPG journals * PubMed * Google Scholar * Alexander G Robling Search for this author in: * NPG journals * PubMed * Google Scholar Supplementary information * Abstract * Author information * Supplementary information PDF files * Supplementary Text and Figures (9M) Supplementary Figures 1–10, Supplementary Tables 1–4 and Supplementary Methods Additional data
• PGRPs kill with an ancient weapon
  - Nat Med 17(6):665-666 (2011)
  Article preview View full access options Nature Medicine | Article Lrp5 functions in bone to regulate bone mass * Yajun Cui1, 2 * Paul J Niziolek3, 4, 5 * Bryan T MacDonald6, 7 * Cassandra R Zylstra8 * Natalia Alenina9 * Daniel R Robinson8 * Zhendong Zhong8 * Susann Matthes9 * Christina M Jacobsen1 * Ronald A Conlon2 * Robert Brommage10 * Qingyun Liu10 * Faika Mseeh10 * David R Powell10 * Qi M Yang10 * Brian Zambrowicz10 * Han Gerrits11 * Jan A Gossen11 * Xi He6, 7 * Michael Bader9 * Bart O Williams8 * Matthew L Warman1, 12, 13 * Alexander G Robling4, 5 * Affiliations * Contributions * Corresponding authorJournal name:Nature MedicineVolume: 17,Pages:684–691Year published:(2011)DOI:doi:10.1038/nm.2388Received14 December 2010Accepted27 April 2011Published online22 May 2011 Abstract * Abstract * Author information * Supplementary information Article tools * Full text * 日本語要約 * Print * Email * Download citation * Order reprints * Rights and permissions * Share/bookmark * Connotea * CiteULike * Facebook * Twitter * Delicious * Digg The human skeleton is affected by mutations in low-density lipoprotein receptor-related protein 5 (LRP5). To understand how LRP5 influences bone properties, we generated mice with osteocyte-specific expression of inducible Lrp5 mutations that cause high and low bone mass phenotypes in humans. We found that bone properties in these mice were comparable to bone properties in mice with inherited mutations. We also induced an Lrp5 mutation in cells that form the appendicular skeleton but not in cells that form the axial skeleton; we observed that bone properties were altered in the limb but not in the spine. These data indicate that Lrp5 signaling functions locally, and they suggest that increasing LRP5 signaling in mature bone cells may be a strategy for treating human disorders associated with low bone mass, such as osteoporosis. Figures at a glance * Figure 1: Generation and characterization of HBM-causing Lrp5 knock-in mice. () Schematic depicting the targeting and genotyping strategies for Lrp5 HBM alleles. The 5′ targeting arm contains a neomycin-resistance cassette (NeoR) flanked by loxP sites (arrowheads). The 3′ targeting arm begins in intron 2 and extends into intron 4, and is followed by a thymidine kinase (TK) cassette. Site-directed mutagenesis altered specific amino acid residues encoded by Lrp5 exon 3 (asterisks). The relative locations and orientation of primers used for PCR genotyping and their expected amplimer sizes are noted. () Autoradiographs of a northern blot containing whole-bone total RNA from mice with different Lrp5 genotypes initially hybridized with a radioactive Lrp5 cDNA probe (top) and subsequently with a glyceraldehyde 3-phosphate dehydrogenase (Gapdh) cDNA probe which serves as a loading control (bottom). () Photograph of an agarose gel containing PCR amplimers derived from genomic DNA of mice with different Lrp5 genotypes. () Graphs depicting the areal bone mi! neral density (aBMD) measured by dual-energy X-ray absorptiometry (DEXA) in mice with different Lrp5 genotypes; mice followed until the age of 16.5 weeks (top and middle). Graphs depicting the percentage of trabecular bone volume in the total volume (BV/TV) of the distal femora and fifth lumbar vertebrae of 16.5-week-old male and female mice with different Lrp5 genotypes (bottom). () Representative μCT scan images obtained from 16.5-week-old mice with different Lrp5 genotypes. Scale bars, 1 mm. () Graphs depicting biomechanical properties of whole femora in a three-point bending assay from 16.5-week-old mice with different Lrp5 genotypes. () Representative images of new bone formation assessed by double calcein labeling of mice with different Lrp5 genotypes. Bone formation rates/bone surface area (BFR/BS); mineral apposition rates (MAR); mineralizing surface/bone surface (MS/BS). The numbers of mice studied are indicated. Error bars show means ± s.d. *P < 0.05 versus WT m! ice; #P < 0.05 versus heterozygous Lrp5 HBM mice. * Figure 2: Effect of activating Lrp5 NeoR-containing HBM alleles. () Graphs depicting femoral and vertebral trabecular BV/TV in WT mice and in mice with Lrp5 NeoR-containing HBM alleles. (,) Graphs depicting femoral trabecular BV/TV () and vertebral trabecular BV/TV () in mice with (shaded bars) and without (unshaded bars) inherited NeoR-containing Lrp5 HBM alleles (+/AN or +/GN, and +/A or +/G, respectively), and with and without Vil1-Cre (+V and –V, respectively) or Dmp1-Cre (+D and –D, respectively) transgenes. () Photographs of agarose gels containing PCR amplimers derived from mouse genomic DNA extracted from either duodenum or femur cortex of mice with different Lrp5 and Cre-transgene genotypes. PCR amplimers correspond to the sizes depicted in Figure 1a. Top, amplimers from the Dmp1-Cre cross. Bottom, amplimers from the Vil1-Cre cross. WT allele (arrowheads), AN or GN allele (double arrowheads) A or G allele (arrows). () Graphs depicting fluorochrome-derived bone formation parameters in the distal femur from 9-week-old female mi! ce that were administered double calcein labeling. Group notations (x axis) follow those described for panel . () Graphs depicting the proportion of distal femur trabecular bone surface covered by osteoclasts (Oc.S; left) and osteoblasts (Ob.S; right). () Graphs depicting femoral and vertebral trabecular BV/TV in 12-week-old mice with (shaded bars) and without (unshaded bars) inherited NeoR-containing Lrp5 HBM alleles (+/AN and +/A), respectively, and with and without the Prrx1-Cre transgene (+P and −P, respectively). MS/BS, MAR, BFR/BS are as defined in Figure 1. The numbers of mice studied are indicated. Error bars show means ± s.d. *P < 0.05 versus NeoR-containing littermates that did not inherit a Cre transgene. * Figure 3: Generation and characterization of mice with a conditional knockout allele of Lrp5. () Schematic depicting the creation of the Lrp5 floxed allele. loxP sites (arrowheads), the neomycin-resistance cassette (NeoR), flippase (Flp) recognition target (FRT) sites (diamonds) and the diphtheria toxin (DT) cassette are shown. The relative locations and orientation of the three primers (arrows) used for PCR genotyping and their expected amplimer sizes are noted. () Photograph of agarose gel depicting PCR amplimers for WT (+), floxed (f) and knockout (−) Lrp5 alleles from genomic DNA of mice with different Lrp5 genotypes. () Photographs of agarose gels containing PCR amplimers derived from mouse genomic DNA extracted from either duodenum or femur cortex of floxed Lrp5 mice, with or without the Dmp1-Cre transgene (top) and with or without the Vil1-Cre transgene (bottom). () Graphs depicting whole-body aBMD (left), femoral trabecular BV/TV (middle) and vertebral trabecular BV/TV (right) in 16-week-old mice homozygous for WT or floxed Lrp5 alleles, with or without the! Dmp1-Cre transgene (+D and –D, respectively). () Graphs depicting whole-body aBMD and tibial trabecular BV/TV in 3-month-old and 12-month-old mice heterozygous or homozygous for floxed Lrp5 alleles with or without the Vil1-Cre transgene (+V and –V, respectively). The numbers of mice studied are indicated. Error bars show means ± s.d. *P < 0.05 versus floxed Lrp5 littermates that did not inherit the Cre transgene. * Figure 4: Effect of Lrp5 genotype on 5HT concentration and on Tph1 expression. () Graphs depicting whole-blood 5HT measured by HPLC in 6-month-old Lrp5 WT and knockout mice that had been backcrossed to C57BL6/J mice. Shown are 5HT measurements in Lrp5 WT and HBM-causing knock-in (G/G) mice on a mixed 129Sv/C57BL/6J background (far left); in 3-month-old Lrp5 WT, knockout and HBM knock-in (+/A) mice on a mixed 129Sv/C57BL/6J genetic background (middle left); and in 3-month-old male (middle right) and 13-month-old female (far right) WT and knockout littermates on a mixed 129SvEvBrd/ C57BL/6J-Tyrc-Brd background. () Graphs depicting the quantity of 5HT extracted from several regions of the intestine, beginning in the duodenum and proceeding through the jejunum, ileum and proximal colon in 3-month-old male (left) and in 13-month-old female (right) Lrp5 WT and knockout littermates on a mixed 129SvEvBrd/ C57BL/6J-Tyrc-Brd background. () Scattergram depicting vertebral trabecular BV/TV and whole blood 5HT measurements in individual Lrp5 WT (open symbols) and k! nockout (filled symbols) littermates. Correlations between BV/TV and whole blood serotonin were r2 = 0.13 (P = 0.16) for male mice, and r2 = 0.02 (P = 0.53) for female mice. () Graphs depicting normalized Tph1 transcript levels in duodenum RNA extracts from Lrp5 WT, knockout, and HBM-causing knock-in (+/A) mice on a mixed 129Sv/C57BL/6J genetic background (left) and duodenum and colon RNA extracts from Lrp5 WT and knockout mice on a 129SvEvBrd/ C57BL/6J-Tyrc-Brd background (right) with Gapdh serving as the internal control. The mean Tph1 expression level for Lrp5 WT duodenum is set as 100%. The numbers of mice studied are indicated. Error bars show means ± s.d. *P < 0.05 versus WT mice. * Figure 5: Bone mass in WT and Tph1−/− mice. () Graphs depicting femoral trabecular BV/TV in WT and Tph1−/− mice on either FVB/N or C57BL/6 backgrounds (left) or on a mixed 129SvEvBrd/C57BL/6J-Tyrc-Brd background (right). () Graphs depicting the vertebral trabecular BV/TV of the fifth lumbar vertebrae in the same mice described in panel . () Graphs depicting lumbar spine aBMD, as measured by DEXA, in the same mice described in panel . The numbers of mice studied are indicated. Error bars show means ± s.d. *P < 0.05 versus WT mice using an unpaired t test; none of these differences remain significant after correcting for multiple testing. * Figure 6: Bone mass after pharmacologic inhibition of Tph1 activity. () Graph depicting dose-dependent changes in 5HT content, compared to vehicle-treated controls, in 9-week-old WT female C57BL/6 mice after receiving daily doses of LP-923941 for 7 days. A daily dose of 250 mg kg−1 lowered 5HT content in whole blood and in intestine, but not in brain. () Graph depicting changes in the intestinal 5HT content in sham-operated (SHM) and ovariectomized (OVX) mice that received vehicle or LP-923941 (250 mg per kg per day) for 6 weeks. Ovariectomy alone reduced 5HT content in the duodenum and colon by ~12% (P < 0.05) compared with SHM mice. Treatment with LP-923941 significantly reduced serotonin content equally in all regions of the intestine in SHM and in OVX mice. () Effect of treatment with LP-923941 (250 mg per kg per day) or teriparatide (80 μg per kg per day), which is the 1–34 residue amino-terminal fragment of human parathyroid hormone (PTH), on serum P1NP levels (left), vertebral trabecular BV/TV (middle left), midshaft femur cortica! l thickness (Ct.th; middle right) and midshaft femoral volumetric BMD (right) in SHM and in OVX mice. () Effect of treating SHM and OVX rats with LP-923941 (50 or 250 mg per kg per day) or teriparatide (80 μg per kg per day) for 6 weeks on jejunal 5HT content (left), femoral trabecular BV/TV (middle) and vertebral trabecular BV/TV (right). The numbers of mice studied are indicated. Error bars show means ± s.d. *P < 0.05 versus vehicle-treated mice. Article preview Read the full article * Instant access to this article: US$32 Buy now * Subscribe to Nature Medicine for full access: Subscribe * Personal subscribers: Log in Additional access options: * Login via Athens * Login via your Institution * Purchase a site license * Use a document delivery service * Rent this article from DeepDyve * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. Author information * Abstract * Author information * Supplementary information Affiliations * Orthopedic Research Laboratories, Department of Orthopedic Surgery, Children's Hospital, Boston, Massachusetts, USA. * Yajun Cui, * Christina M Jacobsen & * Matthew L Warman * Department of Genetics, Case Western Reserve University School of Medicine, Cleveland, Ohio, USA. * Yajun Cui & * Ronald A Conlon * Weldon School of Biomedical Engineering, Purdue University, West Lafayette, Indiana, USA. * Paul J Niziolek * Department of Anatomy & Cell Biology, Indiana University School of Medicine, Indianapolis, Indiana, USA. * Paul J Niziolek & * Alexander G Robling * Department of Biomedical Engineering, Indiana University School of Medicine, Indianapolis, Indiana, USA. * Paul J Niziolek & * Alexander G Robling * F.M. Kirby Neurobiology Center, Children's Hospital, Boston, Massachusetts, USA. * Bryan T MacDonald & * Xi He * Department of Neurology, Harvard Medical School, Boston, Massachusetts, USA. * Bryan T MacDonald & * Xi He * Center for Skeletal Disease Research, Van Andel Research Institute, Grand Rapids, Michigan, USA. * Cassandra R Zylstra, * Daniel R Robinson, * Zhendong Zhong & * Bart O Williams * Max Delbrück Center for Molecular Medicine, Berlin-Buch, Germany. * Natalia Alenina, * Susann Matthes & * Michael Bader * Lexicon Pharmaceuticals, The Woodlands, Texas, USA. * Robert Brommage, * Qingyun Liu, * Faika Mseeh, * David R Powell, * Qi M Yang & * Brian Zambrowicz * Merck Sharp & Dohme Research Laboratories, Oss, The Netherlands. * Han Gerrits & * Jan A Gossen * Howard Hughes Medical Institute, Children's Hospital, Boston, Massachusetts, USA. * Matthew L Warman * Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA. * Matthew L Warman Contributions Y.C. created and did studies on the mice with the Lrp5 HBM alleles and measured serum serotonin levels by competitive ELISA. P.J.N. did radiographic imaging and biomechanical testing on the mice with HBM-associated alleles. B.T.M. contributed to the serotonin and Tph1 qRT-PCR measurements in HBM-causing and Lrp5 knockout mice. C.R.Z. did multiple studies using the conditional Lrp5 knockout mice. N.A. studied the Tph1−/− mice, and with S.M. measured whole blood serotonin levels from HBM-causing and Lrp5-knockout mice by HPLC. D.R.R. generated the conditional Lrp5 knockout strain and Z.Z. participated in conditional inactivation of this allele using different Cre transgenes. C.M.J. carried out the Prrx1-Cre experiments. R.B., F.M. and Q.M.Y. organized studies on Lrp5- and Tph -knockout mice, and also organized the mouse pharmacology experiment. H.G. and J.A.G. organized the rat pharmacology experiment. R.A.C., X.H., M.B., D.R.P., Q.L., B.Z., B.O.W., A.G.R. and M.L.W. desig! ned experiments and provided reagents and financial support. M.L.W. prepared the first draft of the manuscript. All co-authors contributed detailed methods and results, and revised and approved the manuscript. Competing financial interests Employees of Lexicon Pharmaceuticals (R.B., Q.L., F.M., D.R.P., Q.M.Y. and B.Z.) and Merck Sharp & Dohme Research Laboratories (H.G. and J.A.G.) have received compensation in the form of salary and stock options. Corresponding author Correspondence to: * Matthew L Warman Author Details * Yajun Cui Search for this author in: * NPG journals * PubMed * Google Scholar * Paul J Niziolek Search for this author in: * NPG journals * PubMed * Google Scholar * Bryan T MacDonald Search for this author in: * NPG journals * PubMed * Google Scholar * Cassandra R Zylstra Search for this author in: * NPG journals * PubMed * Google Scholar * Natalia Alenina Search for this author in: * NPG journals * PubMed * Google Scholar * Daniel R Robinson Search for this author in: * NPG journals * PubMed * Google Scholar * Zhendong Zhong Search for this author in: * NPG journals * PubMed * Google Scholar * Susann Matthes Search for this author in: * NPG journals * PubMed * Google Scholar * Christina M Jacobsen Search for this author in: * NPG journals * PubMed * Google Scholar * Ronald A Conlon Search for this author in: * NPG journals * PubMed * Google Scholar * Robert Brommage Search for this author in: * NPG journals * PubMed * Google Scholar * Qingyun Liu Search for this author in: * NPG journals * PubMed * Google Scholar * Faika Mseeh Search for this author in: * NPG journals * PubMed * Google Scholar * David R Powell Search for this author in: * NPG journals * PubMed * Google Scholar * Qi M Yang Search for this author in: * NPG journals * PubMed * Google Scholar * Brian Zambrowicz Search for this author in: * NPG journals * PubMed * Google Scholar * Han Gerrits Search for this author in: * NPG journals * PubMed * Google Scholar * Jan A Gossen Search for this author in: * NPG journals * PubMed * Google Scholar * Xi He Search for this author in: * NPG journals * PubMed * Google Scholar * Michael Bader Search for this author in: * NPG journals * PubMed * Google Scholar * Bart O Williams Search for this author in: * NPG journals * PubMed * Google Scholar * Matthew L Warman Contact Matthew L Warman Search for this author in: * NPG journals * PubMed * Google Scholar * Alexander G Robling Search for this author in: * NPG journals * PubMed * Google Scholar Supplementary information * Abstract * Author information * Supplementary information PDF files * Supplementary Text and Figures (9M) Supplementary Figures 1–10, Supplementary Tables 1–4 and Supplementary Methods Additional data
• Tracing the roots of squamous cell carcinomas
  - Nat Med 17(6):668-669 (2011)
  Article preview View full access options Nature Medicine | News Market overlap points to irresponsible use of tuberculosis drugs * Erika Check-HaydenJournal name:Nature MedicineVolume: 17,Page:635Year published:(2011)DOI:doi:10.1038/nm0611-635Published online06 June 2011 sy5/sy5/ZUMA Press/Newscom The private drug market can treat two thirds of those who develop TB. People with active tuberculosis infections turn to the private market for treatment far more often than anyone had realized. And when they do, they encounter a chaotic array of treatment choices, many of which do not meet guidelines drawn up by the World Health Organization. These are the conclusions of a paper published on 4 May that counters the prevailing wisdom that the vast majority of people with tuberculosis are treated through publicly funded programs. Article preview Read the full article * Instant access to this article: US$32 Buy now * Subscribe to Nature Medicine for full access: Subscribe * Personal subscribers: Log in Additional access options: * Login via Athens * Login via your Institution * Purchase a site license * Use a document delivery service * Rent this article from DeepDyve * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. Additional data Author Details * Erika Check-Hayden Search for this author in: * NPG journals * PubMed * Google Scholar
• Research Highlights
  - Nat Med 17(6):670-671 (2011)
  Article preview View full access options Nature Medicine | News Market overlap points to irresponsible use of tuberculosis drugs * Erika Check-HaydenJournal name:Nature MedicineVolume: 17,Page:635Year published:(2011)DOI:doi:10.1038/nm0611-635Published online06 June 2011 sy5/sy5/ZUMA Press/Newscom The private drug market can treat two thirds of those who develop TB. People with active tuberculosis infections turn to the private market for treatment far more often than anyone had realized. And when they do, they encounter a chaotic array of treatment choices, many of which do not meet guidelines drawn up by the World Health Organization. These are the conclusions of a paper published on 4 May that counters the prevailing wisdom that the vast majority of people with tuberculosis are treated through publicly funded programs. Article preview Read the full article * Instant access to this article: US$32 Buy now * Subscribe to Nature Medicine for full access: Subscribe * Personal subscribers: Log in Additional access options: * Login via Athens * Login via your Institution * Purchase a site license * Use a document delivery service * Rent this article from DeepDyve * British Library Document Supply Centre * Infotrieve * Thompson ISI Document Delivery * You can also request this document from your local library through inter-library loan services. Additional data Author Details * Erika Check-Hayden Search for this author in: * NPG journals * PubMed * Google Scholar
• Identification of T helper type 1–like, Foxp3+ regulatory T cells in human autoimmune disease
  - Nat Med 17(6):673-675 (2011)
  Nature Medicine | Brief Communication Identification of T helper type 1–like, Foxp3+ regulatory T cells in human autoimmune disease * Margarita Dominguez-Villar1, 2 * Clare M Baecher-Allan2, 3 * David A Hafler1, 2, 3 * Affiliations * Contributions * Corresponding authorJournal name:Nature MedicineVolume: 17,Pages:673–675Year published:(2011)DOI:doi:10.1038/nm.2389Received09 November 2010Accepted28 April 2010Published online03 May 2011 Abstract * Abstract * Author information * Supplementary information Article tools * Full text * 日本語要約 * Print * Email * Download PDF * Download citation * Order reprints * Rights and permissions * Share/bookmark * Connotea * CiteULike * Facebook * Twitter * Delicious * Digg CD4+CD25highCD127low/– forkhead box p3 (Foxp3)+ regulatory T cells (Treg cells) possess functional plasticity. Here we describe a higher frequency of T helper type 1 (TH1)-like, interferon-γ (IFN-γ)-secreting Foxp3+ T cells in untreated subjects with relapsing remitting multiple sclerosis (RRMS) as compared to healthy control individuals. In subjects treated with IFN-β, the frequency of IFN-γ+Foxp3+ T cells is similar to that in healthy control subjects. In vitro, human Treg cells from healthy subjects acquire a TH1-like phenotype when cultured in the presence of interleukin-12 (IL-12). TH1-like Treg cells show reduced suppressive activity in vitro, which can partially be reversed by IFN-γ–specific antibodies or by removal of IL-12. View full text Figures at a glance * Figure 1: Treg cells from individuals with RRMS secrete IFN-γex vivo. () The frequency of FACS-sorted IFN-γ+ and IL-17+ Treg cells in healthy control individuals (left) and untreated individuals with RRMS (middle, n = 17) gated on Foxp3+ Treg cells. Right, purity analysis of the sorted IFN-γ+Foxp3+ and IFN-γ−Foxp3+ populations from subjects with RRMS used for methylation analysis in . () Percentage of IFN-γ+Foxp3+ and IL-17+Foxp3+ Treg cells (n = 17) as a proportion of total Foxp3+ Treg cells. () Representative example of methylation analysis of the TSDR region of the FOXP3 locus in sorted IFN-γ+Foxp3+ and IFN-γ−Foxp3+ Treg cells from subjects with RRMS. An analysis of IFN-γ+Foxp3− memory T cells from subjects with RRMS is shown as a control. () Proliferation of responder T (Tresp) cells cultured with ex vivo FACS-sorted Treg cells from healthy control subjects and untreated subjects with multiple sclerosis (MS; Treg cell:Tresp cell ratio of 1:2) in the presence or absence of an IFN-γ–specific antibody (n = 4). () The frequency! of IFN-γ+ and IL-17+ Treg cells in healthy control subjects (left) or IFN-β–treated patients with RRMS (right) as assessed by intracellular cytokine staining and FACS analysis. The bar diagram (right) shows the percentage of IFN-γ+Foxp3+ and IL-17+Foxp3+ cells as a proportion of total Foxp3+ Treg cells in healthy controls or IFN-β–treated patients with RRMS (n = 12). Approval for studies was obtained from the Brigham and Women's Hospital Institutional Review Board, and informed consent was obtained from all donors. * Figure 2: Characterization of IL-12–driven, IFN-γ+Foxp3+ Treg cells in vitro from healthy controls. () Intracellular staining for IFN-γ, IL-4, IL-17 and IL-10 of untreated (upper row) and IL-12-stimulated (bottom row) human Treg cells from healthy controls at day 4. () mRNA expression of FOXP3 (left), GATA3 (middle) and TBX21 (right) in Treg cells stimulated in the presence or absence of IL-12 for 5 d (data are a representative example of three experiments performed with similar results; *P < 0.05). () Proliferation of Tresp cells cocultured for 3 d with Treg cells (Treg cell:Tresp cell ratio of 1:2) previously treated with IL-2 (top) or IL-2 + IL-12 (bottom), as assessed by carboxyfluorescein succinimidyl ester (CFSE) dilution. Histograms depict unstimulated Tresp cells alone (top left), Tresp cells stimulated with antibody to CD3 (anti-CD3) and antigen-presenting cells without Treg cells (bottom left) and Tresp cell and Treg cell cocultures without blocking antibodies (second column), in the presence of an IL-10–specific blocking antibody (anti–IL-10; third column),! IFN-γ–specific blocking antibody (anti–IFN-γ; fourth column) or both antibodies (fifth column). () Representative example of staining for TH1-associated chemokines (CCR5 and CXCR3) and TGF-β on IFN-γ+Foxp3+ and IFN-γ−Foxp3+ Treg cells (data are representative of three experiments). Author information * Abstract * Author information * Supplementary information Primary authors * These authors contributed equally to this work. * Clare M Baecher-Allan & * David A Hafler Affiliations * Departments of Neurology and Immunobiology, Yale School of Medicine, New Haven, Connecticut, USA. * Margarita Dominguez-Villar & * David A Hafler * Harvard Medical School, Boston, Massachusetts, USA. * Margarita Dominguez-Villar, * Clare M Baecher-Allan & * David A Hafler Contributions M.D.-V. designed and performed the experiments, analyzed data and wrote the manuscript; C.M.B.-A. and D.A.H. supervised the study and wrote the manuscript. Competing financial interests The authors declare no competing financial interests. Corresponding author Correspondence to: * David A Hafler Author Details * Margarita Dominguez-Villar Search for this author in: * NPG journals * PubMed * Google Scholar * Clare M Baecher-Allan Search for this author in: * NPG journals * PubMed * Google Scholar * David A Hafler Contact David A Hafler Search for this author in: * NPG journals * PubMed * Google Scholar Supplementary information * Abstract * Author information * Supplementary information PDF files * Supplementary Text and Figures (561K) Supplementary Figures 1–15, Supplementary Table 1 and Supplementary Methods Additional data
• Peptidoglycan recognition proteins kill bacteria by activating protein-sensing two-component systems
  - Nat Med 17(6):676-683 (2011)
  Nature Medicine | Brief Communication Identification of T helper type 1–like, Foxp3+ regulatory T cells in human autoimmune disease * Margarita Dominguez-Villar1, 2 * Clare M Baecher-Allan2, 3 * David A Hafler1, 2, 3 * Affiliations * Contributions * Corresponding authorJournal name:Nature MedicineVolume: 17,Pages:673–675Year published:(2011)DOI:doi:10.1038/nm.2389Received09 November 2010Accepted28 April 2010Published online03 May 2011 Abstract * Abstract * Author information * Supplementary information Article tools * Full text * 日本語要約 * Print * Email * Download PDF * Download citation * Order reprints * Rights and permissions * Share/bookmark * Connotea * CiteULike * Facebook * Twitter * Delicious * Digg CD4+CD25highCD127low/– forkhead box p3 (Foxp3)+ regulatory T cells (Treg cells) possess functional plasticity. Here we describe a higher frequency of T helper type 1 (TH1)-like, interferon-γ (IFN-γ)-secreting Foxp3+ T cells in untreated subjects with relapsing remitting multiple sclerosis (RRMS) as compared to healthy control individuals. In subjects treated with IFN-β, the frequency of IFN-γ+Foxp3+ T cells is similar to that in healthy control subjects. In vitro, human Treg cells from healthy subjects acquire a TH1-like phenotype when cultured in the presence of interleukin-12 (IL-12). TH1-like Treg cells show reduced suppressive activity in vitro, which can partially be reversed by IFN-γ–specific antibodies or by removal of IL-12. View full text Figures at a glance * Figure 1: Treg cells from individuals with RRMS secrete IFN-γex vivo. () The frequency of FACS-sorted IFN-γ+ and IL-17+ Treg cells in healthy control individuals (left) and untreated individuals with RRMS (middle, n = 17) gated on Foxp3+ Treg cells. Right, purity analysis of the sorted IFN-γ+Foxp3+ and IFN-γ−Foxp3+ populations from subjects with RRMS used for methylation analysis in . () Percentage of IFN-γ+Foxp3+ and IL-17+Foxp3+ Treg cells (n = 17) as a proportion of total Foxp3+ Treg cells. () Representative example of methylation analysis of the TSDR region of the FOXP3 locus in sorted IFN-γ+Foxp3+ and IFN-γ−Foxp3+ Treg cells from subjects with RRMS. An analysis of IFN-γ+Foxp3− memory T cells from subjects with RRMS is shown as a control. () Proliferation of responder T (Tresp) cells cultured with ex vivo FACS-sorted Treg cells from healthy control subjects and untreated subjects with multiple sclerosis (MS; Treg cell:Tresp cell ratio of 1:2) in the presence or absence of an IFN-γ–specific antibody (n = 4). () The frequency! of IFN-γ+ and IL-17+ Treg cells in healthy control subjects (left) or IFN-β–treated patients with RRMS (right) as assessed by intracellular cytokine staining and FACS analysis. The bar diagram (right) shows the percentage of IFN-γ+Foxp3+ and IL-17+Foxp3+ cells as a proportion of total Foxp3+ Treg cells in healthy controls or IFN-β–treated patients with RRMS (n = 12). Approval for studies was obtained from the Brigham and Women's Hospital Institutional Review Board, and informed consent was obtained from all donors. * Figure 2: Characterization of IL-12–driven, IFN-γ+Foxp3+ Treg cells in vitro from healthy controls. () Intracellular staining for IFN-γ, IL-4, IL-17 and IL-10 of untreated (upper row) and IL-12-stimulated (bottom row) human Treg cells from healthy controls at day 4. () mRNA expression of FOXP3 (left), GATA3 (middle) and TBX21 (right) in Treg cells stimulated in the presence or absence of IL-12 for 5 d (data are a representative example of three experiments performed with similar results; *P < 0.05). () Proliferation of Tresp cells cocultured for 3 d with Treg cells (Treg cell:Tresp cell ratio of 1:2) previously treated with IL-2 (top) or IL-2 + IL-12 (bottom), as assessed by carboxyfluorescein succinimidyl ester (CFSE) dilution. Histograms depict unstimulated Tresp cells alone (top left), Tresp cells stimulated with antibody to CD3 (anti-CD3) and antigen-presenting cells without Treg cells (bottom left) and Tresp cell and Treg cell cocultures without blocking antibodies (second column), in the presence of an IL-10–specific blocking antibody (anti–IL-10; third column),! IFN-γ–specific blocking antibody (anti–IFN-γ; fourth column) or both antibodies (fifth column). () Representative example of staining for TH1-associated chemokines (CCR5 and CXCR3) and TGF-β on IFN-γ+Foxp3+ and IFN-γ−Foxp3+ Treg cells (data are representative of three experiments). Author information * Abstract * Author information * Supplementary information Primary authors * These authors contributed equally to this work. * Clare M Baecher-Allan & * David A Hafler Affiliations * Departments of Neurology and Immunobiology, Yale School of Medicine, New Haven, Connecticut, USA. * Margarita Dominguez-Villar & * David A Hafler * Harvard Medical School, Boston, Massachusetts, USA. * Margarita Dominguez-Villar, * Clare M Baecher-Allan & * David A Hafler Contributions M.D.-V. designed and performed the experiments, analyzed data and wrote the manuscript; C.M.B.-A. and D.A.H. supervised the study and wrote the manuscript. Competing financial interests The authors declare no competing financial interests. Corresponding author Correspondence to: * David A Hafler Author Details * Margarita Dominguez-Villar Search for this author in: * NPG journals * PubMed * Google Scholar * Clare M Baecher-Allan Search for this author in: * NPG journals * PubMed * Google Scholar * David A Hafler Contact David A Hafler Search for this author in: * NPG journals * PubMed * Google Scholar Supplementary information * Abstract * Author information * Supplementary information PDF files * Supplementary Text and Figures (561K) Supplementary Figures 1–15, Supplementary Table 1 and Supplementary Methods Additional data
• Lrp5 functions in bone to regulate bone mass
  - Nat Med 17(6):684-691 (2011)
  Nature Medicine | Brief Communication Identification of T helper type 1–like, Foxp3+ regulatory T cells in human autoimmune disease * Margarita Dominguez-Villar1, 2 * Clare M Baecher-Allan2, 3 * David A Hafler1, 2, 3 * Affiliations * Contributions * Corresponding authorJournal name:Nature MedicineVolume: 17,Pages:673–675Year published:(2011)DOI:doi:10.1038/nm.2389Received09 November 2010Accepted28 April 2010Published online03 May 2011 Abstract * Abstract * Author information * Supplementary information Article tools * Full text * 日本語要約 * Print * Email * Download PDF * Download citation * Order reprints * Rights and permissions * Share/bookmark * Connotea * CiteULike * Facebook * Twitter * Delicious * Digg CD4+CD25highCD127low/– forkhead box p3 (Foxp3)+ regulatory T cells (Treg cells) possess functional plasticity. Here we describe a higher frequency of T helper type 1 (TH1)-like, interferon-γ (IFN-γ)-secreting Foxp3+ T cells in untreated subjects with relapsing remitting multiple sclerosis (RRMS) as compared to healthy control individuals. In subjects treated with IFN-β, the frequency of IFN-γ+Foxp3+ T cells is similar to that in healthy control subjects. In vitro, human Treg cells from healthy subjects acquire a TH1-like phenotype when cultured in the presence of interleukin-12 (IL-12). TH1-like Treg cells show reduced suppressive activity in vitro, which can partially be reversed by IFN-γ–specific antibodies or by removal of IL-12. View full text Figures at a glance * Figure 1: Treg cells from individuals with RRMS secrete IFN-γex vivo. () The frequency of FACS-sorted IFN-γ+ and IL-17+ Treg cells in healthy control individuals (left) and untreated individuals with RRMS (middle, n = 17) gated on Foxp3+ Treg cells. Right, purity analysis of the sorted IFN-γ+Foxp3+ and IFN-γ−Foxp3+ populations from subjects with RRMS used for methylation analysis in . () Percentage of IFN-γ+Foxp3+ and IL-17+Foxp3+ Treg cells (n = 17) as a proportion of total Foxp3+ Treg cells. () Representative example of methylation analysis of the TSDR region of the FOXP3 locus in sorted IFN-γ+Foxp3+ and IFN-γ−Foxp3+ Treg cells from subjects with RRMS. An analysis of IFN-γ+Foxp3− memory T cells from subjects with RRMS is shown as a control. () Proliferation of responder T (Tresp) cells cultured with ex vivo FACS-sorted Treg cells from healthy control subjects and untreated subjects with multiple sclerosis (MS; Treg cell:Tresp cell ratio of 1:2) in the presence or absence of an IFN-γ–specific antibody (n = 4). () The frequency! of IFN-γ+ and IL-17+ Treg cells in healthy control subjects (left) or IFN-β–treated patients with RRMS (right) as assessed by intracellular cytokine staining and FACS analysis. The bar diagram (right) shows the percentage of IFN-γ+Foxp3+ and IL-17+Foxp3+ cells as a proportion of total Foxp3+ Treg cells in healthy controls or IFN-β–treated patients with RRMS (n = 12). Approval for studies was obtained from the Brigham and Women's Hospital Institutional Review Board, and informed consent was obtained from all donors. * Figure 2: Characterization of IL-12–driven, IFN-γ+Foxp3+ Treg cells in vitro from healthy controls. () Intracellular staining for IFN-γ, IL-4, IL-17 and IL-10 of untreated (upper row) and IL-12-stimulated (bottom row) human Treg cells from healthy controls at day 4. () mRNA expression of FOXP3 (left), GATA3 (middle) and TBX21 (right) in Treg cells stimulated in the presence or absence of IL-12 for 5 d (data are a representative example of three experiments performed with similar results; *P < 0.05). () Proliferation of Tresp cells cocultured for 3 d with Treg cells (Treg cell:Tresp cell ratio of 1:2) previously treated with IL-2 (top) or IL-2 + IL-12 (bottom), as assessed by carboxyfluorescein succinimidyl ester (CFSE) dilution. Histograms depict unstimulated Tresp cells alone (top left), Tresp cells stimulated with antibody to CD3 (anti-CD3) and antigen-presenting cells without Treg cells (bottom left) and Tresp cell and Treg cell cocultures without blocking antibodies (second column), in the presence of an IL-10–specific blocking antibody (anti–IL-10; third column),! IFN-γ–specific blocking antibody (anti–IFN-γ; fourth column) or both antibodies (fifth column). () Representative example of staining for TH1-associated chemokines (CCR5 and CXCR3) and TGF-β on IFN-γ+Foxp3+ and IFN-γ−Foxp3+ Treg cells (data are representative of three experiments). Author information * Abstract * Author information * Supplementary information Primary authors * These authors contributed equally to this work. * Clare M Baecher-Allan & * David A Hafler Affiliations * Departments of Neurology and Immunobiology, Yale School of Medicine, New Haven, Connecticut, USA. * Margarita Dominguez-Villar & * David A Hafler * Harvard Medical School, Boston, Massachusetts, USA. * Margarita Dominguez-Villar, * Clare M Baecher-Allan & * David A Hafler Contributions M.D.-V. designed and performed the experiments, analyzed data and wrote the manuscript; C.M.B.-A. and D.A.H. supervised the study and wrote the manuscript. Competing financial interests The authors declare no competing financial interests. Corresponding author Correspondence to: * David A Hafler Author Details * Margarita Dominguez-Villar Search for this author in: * NPG journals * PubMed * Google Scholar * Clare M Baecher-Allan Search for this author in: * NPG journals * PubMed * Google Scholar * David A Hafler Contact David A Hafler Search for this author in: * NPG journals * PubMed * Google Scholar Supplementary information * Abstract * Author information * Supplementary information PDF files * Supplementary Text and Figures (561K) Supplementary Figures 1–15, Supplementary Table 1 and Supplementary Methods Additional data
• Iduna protects the brain from glutamate excitotoxicity and stroke by interfering with poly(ADP-ribose) polymer-induced cell death
  - Nat Med 17(6):692-699 (2011)
  Nature Medicine | Brief Communication Identification of T helper type 1–like, Foxp3+ regulatory T cells in human autoimmune disease * Margarita Dominguez-Villar1, 2 * Clare M Baecher-Allan2, 3 * David A Hafler1, 2, 3 * Affiliations * Contributions * Corresponding authorJournal name:Nature MedicineVolume: 17,Pages:673–675Year published:(2011)DOI:doi:10.1038/nm.2389Received09 November 2010Accepted28 April 2010Published online03 May 2011 Abstract * Abstract * Author information * Supplementary information Article tools * Full text * 日本語要約 * Print * Email * Download PDF * Download citation * Order reprints * Rights and permissions * Share/bookmark * Connotea * CiteULike * Facebook * Twitter * Delicious * Digg CD4+CD25highCD127low/– forkhead box p3 (Foxp3)+ regulatory T cells (Treg cells) possess functional plasticity. Here we describe a higher frequency of T helper type 1 (TH1)-like, interferon-γ (IFN-γ)-secreting Foxp3+ T cells in untreated subjects with relapsing remitting multiple sclerosis (RRMS) as compared to healthy control individuals. In subjects treated with IFN-β, the frequency of IFN-γ+Foxp3+ T cells is similar to that in healthy control subjects. In vitro, human Treg cells from healthy subjects acquire a TH1-like phenotype when cultured in the presence of interleukin-12 (IL-12). TH1-like Treg cells show reduced suppressive activity in vitro, which can partially be reversed by IFN-γ–specific antibodies or by removal of IL-12. View full text Figures at a glance * Figure 1: Treg cells from individuals with RRMS secrete IFN-γex vivo. () The frequency of FACS-sorted IFN-γ+ and IL-17+ Treg cells in healthy control individuals (left) and untreated individuals with RRMS (middle, n = 17) gated on Foxp3+ Treg cells. Right, purity analysis of the sorted IFN-γ+Foxp3+ and IFN-γ−Foxp3+ populations from subjects with RRMS used for methylation analysis in . () Percentage of IFN-γ+Foxp3+ and IL-17+Foxp3+ Treg cells (n = 17) as a proportion of total Foxp3+ Treg cells. () Representative example of methylation analysis of the TSDR region of the FOXP3 locus in sorted IFN-γ+Foxp3+ and IFN-γ−Foxp3+ Treg cells from subjects with RRMS. An analysis of IFN-γ+Foxp3− memory T cells from subjects with RRMS is shown as a control. () Proliferation of responder T (Tresp) cells cultured with ex vivo FACS-sorted Treg cells from healthy control subjects and untreated subjects with multiple sclerosis (MS; Treg cell:Tresp cell ratio of 1:2) in the presence or absence of an IFN-γ–specific antibody (n = 4). () The frequency! of IFN-γ+ and IL-17+ Treg cells in healthy control subjects (left) or IFN-β–treated patients with RRMS (right) as assessed by intracellular cytokine staining and FACS analysis. The bar diagram (right) shows the percentage of IFN-γ+Foxp3+ and IL-17+Foxp3+ cells as a proportion of total Foxp3+ Treg cells in healthy controls or IFN-β–treated patients with RRMS (n = 12). Approval for studies was obtained from the Brigham and Women's Hospital Institutional Review Board, and informed consent was obtained from all donors. * Figure 2: Characterization of IL-12–driven, IFN-γ+Foxp3+ Treg cells in vitro from healthy controls. () Intracellular staining for IFN-γ, IL-4, IL-17 and IL-10 of untreated (upper row) and IL-12-stimulated (bottom row) human Treg cells from healthy controls at day 4. () mRNA expression of FOXP3 (left), GATA3 (middle) and TBX21 (right) in Treg cells stimulated in the presence or absence of IL-12 for 5 d (data are a representative example of three experiments performed with similar results; *P < 0.05). () Proliferation of Tresp cells cocultured for 3 d with Treg cells (Treg cell:Tresp cell ratio of 1:2) previously treated with IL-2 (top) or IL-2 + IL-12 (bottom), as assessed by carboxyfluorescein succinimidyl ester (CFSE) dilution. Histograms depict unstimulated Tresp cells alone (top left), Tresp cells stimulated with antibody to CD3 (anti-CD3) and antigen-presenting cells without Treg cells (bottom left) and Tresp cell and Treg cell cocultures without blocking antibodies (second column), in the presence of an IL-10–specific blocking antibody (anti–IL-10; third column),! IFN-γ–specific blocking antibody (anti–IFN-γ; fourth column) or both antibodies (fifth column). () Representative example of staining for TH1-associated chemokines (CCR5 and CXCR3) and TGF-β on IFN-γ+Foxp3+ and IFN-γ−Foxp3+ Treg cells (data are representative of three experiments). Author information * Abstract * Author information * Supplementary information Primary authors * These authors contributed equally to this work. * Clare M Baecher-Allan & * David A Hafler Affiliations * Departments of Neurology and Immunobiology, Yale School of Medicine, New Haven, Connecticut, USA. * Margarita Dominguez-Villar & * David A Hafler * Harvard Medical School, Boston, Massachusetts, USA. * Margarita Dominguez-Villar, * Clare M Baecher-Allan & * David A Hafler Contributions M.D.-V. designed and performed the experiments, analyzed data and wrote the manuscript; C.M.B.-A. and D.A.H. supervised the study and wrote the manuscript. Competing financial interests The authors declare no competing financial interests. Corresponding author Correspondence to: * David A Hafler Author Details * Margarita Dominguez-Villar Search for this author in: * NPG journals * PubMed * Google Scholar * Clare M Baecher-Allan Search for this author in: * NPG journals * PubMed * Google Scholar * David A Hafler Contact David A Hafler Search for this author in: * NPG journals * PubMed * Google Scholar Supplementary information * Abstract * Author information * Supplementary information PDF files * Supplementary Text and Figures (561K) Supplementary Figures 1–15, Supplementary Table 1 and Supplementary Methods Additional data
• Alternatively spliced NKp30 isoforms affect the prognosis of gastrointestinal stromal tumors
  - Nat Med 17(6):700-707 (2011)
  Nature Medicine | Brief Communication Identification of T helper type 1–like, Foxp3+ regulatory T cells in human autoimmune disease * Margarita Dominguez-Villar1, 2 * Clare M Baecher-Allan2, 3 * David A Hafler1, 2, 3 * Affiliations * Contributions * Corresponding authorJournal name:Nature MedicineVolume: 17,Pages:673–675Year published:(2011)DOI:doi:10.1038/nm.2389Received09 November 2010Accepted28 April 2010Published online03 May 2011 Abstract * Abstract * Author information * Supplementary information Article tools * Full text * 日本語要約 * Print * Email * Download PDF * Download citation * Order reprints * Rights and permissions * Share/bookmark * Connotea * CiteULike * Facebook * Twitter * Delicious * Digg CD4+CD25highCD127low/– forkhead box p3 (Foxp3)+ regulatory T cells (Treg cells) possess functional plasticity. Here we describe a higher frequency of T helper type 1 (TH1)-like, interferon-γ (IFN-γ)-secreting Foxp3+ T cells in untreated subjects with relapsing remitting multiple sclerosis (RRMS) as compared to healthy control individuals. In subjects treated with IFN-β, the frequency of IFN-γ+Foxp3+ T cells is similar to that in healthy control subjects. In vitro, human Treg cells from healthy subjects acquire a TH1-like phenotype when cultured in the presence of interleukin-12 (IL-12). TH1-like Treg cells show reduced suppressive activity in vitro, which can partially be reversed by IFN-γ–specific antibodies or by removal of IL-12. View full text Figures at a glance * Figure 1: Treg cells from individuals with RRMS secrete IFN-γex vivo. () The frequency of FACS-sorted IFN-γ+ and IL-17+ Treg cells in healthy control individuals (left) and untreated individuals with RRMS (middle, n = 17) gated on Foxp3+ Treg cells. Right, purity analysis of the sorted IFN-γ+Foxp3+ and IFN-γ−Foxp3+ populations from subjects with RRMS used for methylation analysis in . () Percentage of IFN-γ+Foxp3+ and IL-17+Foxp3+ Treg cells (n = 17) as a proportion of total Foxp3+ Treg cells. () Representative example of methylation analysis of the TSDR region of the FOXP3 locus in sorted IFN-γ+Foxp3+ and IFN-γ−Foxp3+ Treg cells from subjects with RRMS. An analysis of IFN-γ+Foxp3− memory T cells from subjects with RRMS is shown as a control. () Proliferation of responder T (Tresp) cells cultured with ex vivo FACS-sorted Treg cells from healthy control subjects and untreated subjects with multiple sclerosis (MS; Treg cell:Tresp cell ratio of 1:2) in the presence or absence of an IFN-γ–specific antibody (n = 4). () The frequency! of IFN-γ+ and IL-17+ Treg cells in healthy control subjects (left) or IFN-β–treated patients with RRMS (right) as assessed by intracellular cytokine staining and FACS analysis. The bar diagram (right) shows the percentage of IFN-γ+Foxp3+ and IL-17+Foxp3+ cells as a proportion of total Foxp3+ Treg cells in healthy controls or IFN-β–treated patients with RRMS (n = 12). Approval for studies was obtained from the Brigham and Women's Hospital Institutional Review Board, and informed consent was obtained from all donors. * Figure 2: Characterization of IL-12–driven, IFN-γ+Foxp3+ Treg cells in vitro from healthy controls. () Intracellular staining for IFN-γ, IL-4, IL-17 and IL-10 of untreated (upper row) and IL-12-stimulated (bottom row) human Treg cells from healthy controls at day 4. () mRNA expression of FOXP3 (left), GATA3 (middle) and TBX21 (right) in Treg cells stimulated in the presence or absence of IL-12 for 5 d (data are a representative example of three experiments performed with similar results; *P < 0.05). () Proliferation of Tresp cells cocultured for 3 d with Treg cells (Treg cell:Tresp cell ratio of 1:2) previously treated with IL-2 (top) or IL-2 + IL-12 (bottom), as assessed by carboxyfluorescein succinimidyl ester (CFSE) dilution. Histograms depict unstimulated Tresp cells alone (top left), Tresp cells stimulated with antibody to CD3 (anti-CD3) and antigen-presenting cells without Treg cells (bottom left) and Tresp cell and Treg cell cocultures without blocking antibodies (second column), in the presence of an IL-10–specific blocking antibody (anti–IL-10; third column),! IFN-γ–specific blocking antibody (anti–IFN-γ; fourth column) or both antibodies (fifth column). () Representative example of staining for TH1-associated chemokines (CCR5 and CXCR3) and TGF-β on IFN-γ+Foxp3+ and IFN-γ−Foxp3+ Treg cells (data are representative of three experiments). Author information * Abstract * Author information * Supplementary information Primary authors * These authors contributed equally to this work. * Clare M Baecher-Allan & * David A Hafler Affiliations * Departments of Neurology and Immunobiology, Yale School of Medicine, New Haven, Connecticut, USA. * Margarita Dominguez-Villar & * David A Hafler * Harvard Medical School, Boston, Massachusetts, USA. * Margarita Dominguez-Villar, * Clare M Baecher-Allan & * David A Hafler Contributions M.D.-V. designed and performed the experiments, analyzed data and wrote the manuscript; C.M.B.-A. and D.A.H. supervised the study and wrote the manuscript. Competing financial interests The authors declare no competing financial interests. Corresponding author Correspondence to: * David A Hafler Author Details * Margarita Dominguez-Villar Search for this author in: * NPG journals * PubMed * Google Scholar * Clare M Baecher-Allan Search for this author in: * NPG journals * PubMed * Google Scholar * David A Hafler Contact David A Hafler Search for this author in: * NPG journals * PubMed * Google Scholar Supplementary information * Abstract * Author information * Supplementary information PDF files * Supplementary Text and Figures (561K) Supplementary Figures 1–15, Supplementary Table 1 and Supplementary Methods Additional data
• Elevated expression of CUEDC2 protein confers endocrine resistance in breast cancer
  - Nat Med 17(6):708-714 (2011)
  Nature Medicine | Brief Communication Identification of T helper type 1–like, Foxp3+ regulatory T cells in human autoimmune disease * Margarita Dominguez-Villar1, 2 * Clare M Baecher-Allan2, 3 * David A Hafler1, 2, 3 * Affiliations * Contributions * Corresponding authorJournal name:Nature MedicineVolume: 17,Pages:673–675Year published:(2011)DOI:doi:10.1038/nm.2389Received09 November 2010Accepted28 April 2010Published online03 May 2011 Abstract * Abstract * Author information * Supplementary information Article tools * Full text * 日本語要約 * Print * Email * Download PDF * Download citation * Order reprints * Rights and permissions * Share/bookmark * Connotea * CiteULike * Facebook * Twitter * Delicious * Digg CD4+CD25highCD127low/– forkhead box p3 (Foxp3)+ regulatory T cells (Treg cells) possess functional plasticity. Here we describe a higher frequency of T helper type 1 (TH1)-like, interferon-γ (IFN-γ)-secreting Foxp3+ T cells in untreated subjects with relapsing remitting multiple sclerosis (RRMS) as compared to healthy control individuals. In subjects treated with IFN-β, the frequency of IFN-γ+Foxp3+ T cells is similar to that in healthy control subjects. In vitro, human Treg cells from healthy subjects acquire a TH1-like phenotype when cultured in the presence of interleukin-12 (IL-12). TH1-like Treg cells show reduced suppressive activity in vitro, which can partially be reversed by IFN-γ–specific antibodies or by removal of IL-12. View full text Figures at a glance * Figure 1: Treg cells from individuals with RRMS secrete IFN-γex vivo. () The frequency of FACS-sorted IFN-γ+ and IL-17+ Treg cells in healthy control individuals (left) and untreated individuals with RRMS (middle, n = 17) gated on Foxp3+ Treg cells. Right, purity analysis of the sorted IFN-γ+Foxp3+ and IFN-γ−Foxp3+ populations from subjects with RRMS used for methylation analysis in . () Percentage of IFN-γ+Foxp3+ and IL-17+Foxp3+ Treg cells (n = 17) as a proportion of total Foxp3+ Treg cells. () Representative example of methylation analysis of the TSDR region of the FOXP3 locus in sorted IFN-γ+Foxp3+ and IFN-γ−Foxp3+ Treg cells from subjects with RRMS. An analysis of IFN-γ+Foxp3− memory T cells from subjects with RRMS is shown as a control. () Proliferation of responder T (Tresp) cells cultured with ex vivo FACS-sorted Treg cells from healthy control subjects and untreated subjects with multiple sclerosis (MS; Treg cell:Tresp cell ratio of 1:2) in the presence or absence of an IFN-γ–specific antibody (n = 4). () The frequency! of IFN-γ+ and IL-17+ Treg cells in healthy control subjects (left) or IFN-β–treated patients with RRMS (right) as assessed by intracellular cytokine staining and FACS analysis. The bar diagram (right) shows the percentage of IFN-γ+Foxp3+ and IL-17+Foxp3+ cells as a proportion of total Foxp3+ Treg cells in healthy controls or IFN-β–treated patients with RRMS (n = 12). Approval for studies was obtained from the Brigham and Women's Hospital Institutional Review Board, and informed consent was obtained from all donors. * Figure 2: Characterization of IL-12–driven, IFN-γ+Foxp3+ Treg cells in vitro from healthy controls. () Intracellular staining for IFN-γ, IL-4, IL-17 and IL-10 of untreated (upper row) and IL-12-stimulated (bottom row) human Treg cells from healthy controls at day 4. () mRNA expression of FOXP3 (left), GATA3 (middle) and TBX21 (right) in Treg cells stimulated in the presence or absence of IL-12 for 5 d (data are a representative example of three experiments performed with similar results; *P < 0.05). () Proliferation of Tresp cells cocultured for 3 d with Treg cells (Treg cell:Tresp cell ratio of 1:2) previously treated with IL-2 (top) or IL-2 + IL-12 (bottom), as assessed by carboxyfluorescein succinimidyl ester (CFSE) dilution. Histograms depict unstimulated Tresp cells alone (top left), Tresp cells stimulated with antibody to CD3 (anti-CD3) and antigen-presenting cells without Treg cells (bottom left) and Tresp cell and Treg cell cocultures without blocking antibodies (second column), in the presence of an IL-10–specific blocking antibody (anti–IL-10; third column),! IFN-γ–specific blocking antibody (anti–IFN-γ; fourth column) or both antibodies (fifth column). () Representative example of staining for TH1-associated chemokines (CCR5 and CXCR3) and TGF-β on IFN-γ+Foxp3+ and IFN-γ−Foxp3+ Treg cells (data are representative of three experiments). Author information * Abstract * Author information * Supplementary information Primary authors * These authors contributed equally to this work. * Clare M Baecher-Allan & * David A Hafler Affiliations * Departments of Neurology and Immunobiology, Yale School of Medicine, New Haven, Connecticut, USA. * Margarita Dominguez-Villar & * David A Hafler * Harvard Medical School, Boston, Massachusetts, USA. * Margarita Dominguez-Villar, * Clare M Baecher-Allan & * David A Hafler Contributions M.D.-V. designed and performed the experiments, analyzed data and wrote the manuscript; C.M.B.-A. and D.A.H. supervised the study and wrote the manuscript. Competing financial interests The authors declare no competing financial interests. Corresponding author Correspondence to: * David A Hafler Author Details * Margarita Dominguez-Villar Search for this author in: * NPG journals * PubMed * Google Scholar * Clare M Baecher-Allan Search for this author in: * NPG journals * PubMed * Google Scholar * David A Hafler Contact David A Hafler Search for this author in: * NPG journals * PubMed * Google Scholar Supplementary information * Abstract * Author information * Supplementary information PDF files * Supplementary Text and Figures (561K) Supplementary Figures 1–15, Supplementary Table 1 and Supplementary Methods Additional data
• Kinome screening for regulators of the estrogen receptor identifies LMTK3 as a new therapeutic target in breast cancer
  - Nat Med 17(6):715-719 (2011)
  Nature Medicine | Brief Communication Identification of T helper type 1–like, Foxp3+ regulatory T cells in human autoimmune disease * Margarita Dominguez-Villar1, 2 * Clare M Baecher-Allan2, 3 * David A Hafler1, 2, 3 * Affiliations * Contributions * Corresponding authorJournal name:Nature MedicineVolume: 17,Pages:673–675Year published:(2011)DOI:doi:10.1038/nm.2389Received09 November 2010Accepted28 April 2010Published online03 May 2011 Abstract * Abstract * Author information * Supplementary information Article tools * Full text * 日本語要約 * Print * Email * Download PDF * Download citation * Order reprints * Rights and permissions * Share/bookmark * Connotea * CiteULike * Facebook * Twitter * Delicious * Digg CD4+CD25highCD127low/– forkhead box p3 (Foxp3)+ regulatory T cells (Treg cells) possess functional plasticity. Here we describe a higher frequency of T helper type 1 (TH1)-like, interferon-γ (IFN-γ)-secreting Foxp3+ T cells in untreated subjects with relapsing remitting multiple sclerosis (RRMS) as compared to healthy control individuals. In subjects treated with IFN-β, the frequency of IFN-γ+Foxp3+ T cells is similar to that in healthy control subjects. In vitro, human Treg cells from healthy subjects acquire a TH1-like phenotype when cultured in the presence of interleukin-12 (IL-12). TH1-like Treg cells show reduced suppressive activity in vitro, which can partially be reversed by IFN-γ–specific antibodies or by removal of IL-12. View full text Figures at a glance * Figure 1: Treg cells from individuals with RRMS secrete IFN-γex vivo. () The frequency of FACS-sorted IFN-γ+ and IL-17+ Treg cells in healthy control individuals (left) and untreated individuals with RRMS (middle, n = 17) gated on Foxp3+ Treg cells. Right, purity analysis of the sorted IFN-γ+Foxp3+ and IFN-γ−Foxp3+ populations from subjects with RRMS used for methylation analysis in . () Percentage of IFN-γ+Foxp3+ and IL-17+Foxp3+ Treg cells (n = 17) as a proportion of total Foxp3+ Treg cells. () Representative example of methylation analysis of the TSDR region of the FOXP3 locus in sorted IFN-γ+Foxp3+ and IFN-γ−Foxp3+ Treg cells from subjects with RRMS. An analysis of IFN-γ+Foxp3− memory T cells from subjects with RRMS is shown as a control. () Proliferation of responder T (Tresp) cells cultured with ex vivo FACS-sorted Treg cells from healthy control subjects and untreated subjects with multiple sclerosis (MS; Treg cell:Tresp cell ratio of 1:2) in the presence or absence of an IFN-γ–specific antibody (n = 4). () The frequency! of IFN-γ+ and IL-17+ Treg cells in healthy control subjects (left) or IFN-β–treated patients with RRMS (right) as assessed by intracellular cytokine staining and FACS analysis. The bar diagram (right) shows the percentage of IFN-γ+Foxp3+ and IL-17+Foxp3+ cells as a proportion of total Foxp3+ Treg cells in healthy controls or IFN-β–treated patients with RRMS (n = 12). Approval for studies was obtained from the Brigham and Women's Hospital Institutional Review Board, and informed consent was obtained from all donors. * Figure 2: Characterization of IL-12–driven, IFN-γ+Foxp3+ Treg cells in vitro from healthy controls. () Intracellular staining for IFN-γ, IL-4, IL-17 and IL-10 of untreated (upper row) and IL-12-stimulated (bottom row) human Treg cells from healthy controls at day 4. () mRNA expression of FOXP3 (left), GATA3 (middle) and TBX21 (right) in Treg cells stimulated in the presence or absence of IL-12 for 5 d (data are a representative example of three experiments performed with similar results; *P < 0.05). () Proliferation of Tresp cells cocultured for 3 d with Treg cells (Treg cell:Tresp cell ratio of 1:2) previously treated with IL-2 (top) or IL-2 + IL-12 (bottom), as assessed by carboxyfluorescein succinimidyl ester (CFSE) dilution. Histograms depict unstimulated Tresp cells alone (top left), Tresp cells stimulated with antibody to CD3 (anti-CD3) and antigen-presenting cells without Treg cells (bottom left) and Tresp cell and Treg cell cocultures without blocking antibodies (second column), in the presence of an IL-10–specific blocking antibody (anti–IL-10; third column),! IFN-γ–specific blocking antibody (anti–IFN-γ; fourth column) or both antibodies (fifth column). () Representative example of staining for TH1-associated chemokines (CCR5 and CXCR3) and TGF-β on IFN-γ+Foxp3+ and IFN-γ−Foxp3+ Treg cells (data are representative of three experiments). Author information * Abstract * Author information * Supplementary information Primary authors * These authors contributed equally to this work. * Clare M Baecher-Allan & * David A Hafler Affiliations * Departments of Neurology and Immunobiology, Yale School of Medicine, New Haven, Connecticut, USA. * Margarita Dominguez-Villar & * David A Hafler * Harvard Medical School, Boston, Massachusetts, USA. * Margarita Dominguez-Villar, * Clare M Baecher-Allan & * David A Hafler Contributions M.D.-V. designed and performed the experiments, analyzed data and wrote the manuscript; C.M.B.-A. and D.A.H. supervised the study and wrote the manuscript. Competing financial interests The authors declare no competing financial interests. Corresponding author Correspondence to: * David A Hafler Author Details * Margarita Dominguez-Villar Search for this author in: * NPG journals * PubMed * Google Scholar * Clare M Baecher-Allan Search for this author in: * NPG journals * PubMed * Google Scholar * David A Hafler Contact David A Hafler Search for this author in: * NPG journals * PubMed * Google Scholar Supplementary information * Abstract * Author information * Supplementary information PDF files * Supplementary Text and Figures (561K) Supplementary Figures 1–15, Supplementary Table 1 and Supplementary Methods Additional data
• Misregulated alternative splicing of BIN1 is associated with T tubule alterations and muscle weakness in myotonic dystrophy
  - Nat Med 17(6):720-725 (2011)
  Nature Medicine | Brief Communication Identification of T helper type 1–like, Foxp3+ regulatory T cells in human autoimmune disease * Margarita Dominguez-Villar1, 2 * Clare M Baecher-Allan2, 3 * David A Hafler1, 2, 3 * Affiliations * Contributions * Corresponding authorJournal name:Nature MedicineVolume: 17,Pages:673–675Year published:(2011)DOI:doi:10.1038/nm.2389Received09 November 2010Accepted28 April 2010Published online03 May 2011 Abstract * Abstract * Author information * Supplementary information Article tools * Full text * 日本語要約 * Print * Email * Download PDF * Download citation * Order reprints * Rights and permissions * Share/bookmark * Connotea * CiteULike * Facebook * Twitter * Delicious * Digg CD4+CD25highCD127low/– forkhead box p3 (Foxp3)+ regulatory T cells (Treg cells) possess functional plasticity. Here we describe a higher frequency of T helper type 1 (TH1)-like, interferon-γ (IFN-γ)-secreting Foxp3+ T cells in untreated subjects with relapsing remitting multiple sclerosis (RRMS) as compared to healthy control individuals. In subjects treated with IFN-β, the frequency of IFN-γ+Foxp3+ T cells is similar to that in healthy control subjects. In vitro, human Treg cells from healthy subjects acquire a TH1-like phenotype when cultured in the presence of interleukin-12 (IL-12). TH1-like Treg cells show reduced suppressive activity in vitro, which can partially be reversed by IFN-γ–specific antibodies or by removal of IL-12. View full text Figures at a glance * Figure 1: Treg cells from individuals with RRMS secrete IFN-γex vivo. () The frequency of FACS-sorted IFN-γ+ and IL-17+ Treg cells in healthy control individuals (left) and untreated individuals with RRMS (middle, n = 17) gated on Foxp3+ Treg cells. Right, purity analysis of the sorted IFN-γ+Foxp3+ and IFN-γ−Foxp3+ populations from subjects with RRMS used for methylation analysis in . () Percentage of IFN-γ+Foxp3+ and IL-17+Foxp3+ Treg cells (n = 17) as a proportion of total Foxp3+ Treg cells. () Representative example of methylation analysis of the TSDR region of the FOXP3 locus in sorted IFN-γ+Foxp3+ and IFN-γ−Foxp3+ Treg cells from subjects with RRMS. An analysis of IFN-γ+Foxp3− memory T cells from subjects with RRMS is shown as a control. () Proliferation of responder T (Tresp) cells cultured with ex vivo FACS-sorted Treg cells from healthy control subjects and untreated subjects with multiple sclerosis (MS; Treg cell:Tresp cell ratio of 1:2) in the presence or absence of an IFN-γ–specific antibody (n = 4). () The frequency! of IFN-γ+ and IL-17+ Treg cells in healthy control subjects (left) or IFN-β–treated patients with RRMS (right) as assessed by intracellular cytokine staining and FACS analysis. The bar diagram (right) shows the percentage of IFN-γ+Foxp3+ and IL-17+Foxp3+ cells as a proportion of total Foxp3+ Treg cells in healthy controls or IFN-β–treated patients with RRMS (n = 12). Approval for studies was obtained from the Brigham and Women's Hospital Institutional Review Board, and informed consent was obtained from all donors. * Figure 2: Characterization of IL-12–driven, IFN-γ+Foxp3+ Treg cells in vitro from healthy controls. () Intracellular staining for IFN-γ, IL-4, IL-17 and IL-10 of untreated (upper row) and IL-12-stimulated (bottom row) human Treg cells from healthy controls at day 4. () mRNA expression of FOXP3 (left), GATA3 (middle) and TBX21 (right) in Treg cells stimulated in the presence or absence of IL-12 for 5 d (data are a representative example of three experiments performed with similar results; *P < 0.05). () Proliferation of Tresp cells cocultured for 3 d with Treg cells (Treg cell:Tresp cell ratio of 1:2) previously treated with IL-2 (top) or IL-2 + IL-12 (bottom), as assessed by carboxyfluorescein succinimidyl ester (CFSE) dilution. Histograms depict unstimulated Tresp cells alone (top left), Tresp cells stimulated with antibody to CD3 (anti-CD3) and antigen-presenting cells without Treg cells (bottom left) and Tresp cell and Treg cell cocultures without blocking antibodies (second column), in the presence of an IL-10–specific blocking antibody (anti–IL-10; third column),! IFN-γ–specific blocking antibody (anti–IFN-γ; fourth column) or both antibodies (fifth column). () Representative example of staining for TH1-associated chemokines (CCR5 and CXCR3) and TGF-β on IFN-γ+Foxp3+ and IFN-γ−Foxp3+ Treg cells (data are representative of three experiments). Author information * Abstract * Author information * Supplementary information Primary authors * These authors contributed equally to this work. * Clare M Baecher-Allan & * David A Hafler Affiliations * Departments of Neurology and Immunobiology, Yale School of Medicine, New Haven, Connecticut, USA. * Margarita Dominguez-Villar & * David A Hafler * Harvard Medical School, Boston, Massachusetts, USA. * Margarita Dominguez-Villar, * Clare M Baecher-Allan & * David A Hafler Contributions M.D.-V. designed and performed the experiments, analyzed data and wrote the manuscript; C.M.B.-A. and D.A.H. supervised the study and wrote the manuscript. Competing financial interests The authors declare no competing financial interests. Corresponding author Correspondence to: * David A Hafler Author Details * Margarita Dominguez-Villar Search for this author in: * NPG journals * PubMed * Google Scholar * Clare M Baecher-Allan Search for this author in: * NPG journals * PubMed * Google Scholar * David A Hafler Contact David A Hafler Search for this author in: * NPG journals * PubMed * Google Scholar Supplementary information * Abstract * Author information * Supplementary information PDF files * Supplementary Text and Figures (561K) Supplementary Figures 1–15, Supplementary Table 1 and Supplementary Methods Additional data
• Defective Wnt-dependent cerebellar midline fusion in a mouse model of Joubert syndrome
  - Nat Med 17(6):726-731 (2011)
  Nature Medicine | Brief Communication Identification of T helper type 1–like, Foxp3+ regulatory T cells in human autoimmune disease * Margarita Dominguez-Villar1, 2 * Clare M Baecher-Allan2, 3 * David A Hafler1, 2, 3 * Affiliations * Contributions * Corresponding authorJournal name:Nature MedicineVolume: 17,Pages:673–675Year published:(2011)DOI:doi:10.1038/nm.2389Received09 November 2010Accepted28 April 2010Published online03 May 2011 Abstract * Abstract * Author information * Supplementary information Article tools * Full text * 日本語要約 * Print * Email * Download PDF * Download citation * Order reprints * Rights and permissions * Share/bookmark * Connotea * CiteULike * Facebook * Twitter * Delicious * Digg CD4+CD25highCD127low/– forkhead box p3 (Foxp3)+ regulatory T cells (Treg cells) possess functional plasticity. Here we describe a higher frequency of T helper type 1 (TH1)-like, interferon-γ (IFN-γ)-secreting Foxp3+ T cells in untreated subjects with relapsing remitting multiple sclerosis (RRMS) as compared to healthy control individuals. In subjects treated with IFN-β, the frequency of IFN-γ+Foxp3+ T cells is similar to that in healthy control subjects. In vitro, human Treg cells from healthy subjects acquire a TH1-like phenotype when cultured in the presence of interleukin-12 (IL-12). TH1-like Treg cells show reduced suppressive activity in vitro, which can partially be reversed by IFN-γ–specific antibodies or by removal of IL-12. View full text Figures at a glance * Figure 1: Treg cells from individuals with RRMS secrete IFN-γex vivo. () The frequency of FACS-sorted IFN-γ+ and IL-17+ Treg cells in healthy control individuals (left) and untreated individuals with RRMS (middle, n = 17) gated on Foxp3+ Treg cells. Right, purity analysis of the sorted IFN-γ+Foxp3+ and IFN-γ−Foxp3+ populations from subjects with RRMS used for methylation analysis in . () Percentage of IFN-γ+Foxp3+ and IL-17+Foxp3+ Treg cells (n = 17) as a proportion of total Foxp3+ Treg cells. () Representative example of methylation analysis of the TSDR region of the FOXP3 locus in sorted IFN-γ+Foxp3+ and IFN-γ−Foxp3+ Treg cells from subjects with RRMS. An analysis of IFN-γ+Foxp3− memory T cells from subjects with RRMS is shown as a control. () Proliferation of responder T (Tresp) cells cultured with ex vivo FACS-sorted Treg cells from healthy control subjects and untreated subjects with multiple sclerosis (MS; Treg cell:Tresp cell ratio of 1:2) in the presence or absence of an IFN-γ–specific antibody (n = 4). () The frequency! of IFN-γ+ and IL-17+ Treg cells in healthy control subjects (left) or IFN-β–treated patients with RRMS (right) as assessed by intracellular cytokine staining and FACS analysis. The bar diagram (right) shows the percentage of IFN-γ+Foxp3+ and IL-17+Foxp3+ cells as a proportion of total Foxp3+ Treg cells in healthy controls or IFN-β–treated patients with RRMS (n = 12). Approval for studies was obtained from the Brigham and Women's Hospital Institutional Review Board, and informed consent was obtained from all donors. * Figure 2: Characterization of IL-12–driven, IFN-γ+Foxp3+ Treg cells in vitro from healthy controls. () Intracellular staining for IFN-γ, IL-4, IL-17 and IL-10 of untreated (upper row) and IL-12-stimulated (bottom row) human Treg cells from healthy controls at day 4. () mRNA expression of FOXP3 (left), GATA3 (middle) and TBX21 (right) in Treg cells stimulated in the presence or absence of IL-12 for 5 d (data are a representative example of three experiments performed with similar results; *P < 0.05). () Proliferation of Tresp cells cocultured for 3 d with Treg cells (Treg cell:Tresp cell ratio of 1:2) previously treated with IL-2 (top) or IL-2 + IL-12 (bottom), as assessed by carboxyfluorescein succinimidyl ester (CFSE) dilution. Histograms depict unstimulated Tresp cells alone (top left), Tresp cells stimulated with antibody to CD3 (anti-CD3) and antigen-presenting cells without Treg cells (bottom left) and Tresp cell and Treg cell cocultures without blocking antibodies (second column), in the presence of an IL-10–specific blocking antibody (anti–IL-10; third column),! IFN-γ–specific blocking antibody (anti–IFN-γ; fourth column) or both antibodies (fifth column). () Representative example of staining for TH1-associated chemokines (CCR5 and CXCR3) and TGF-β on IFN-γ+Foxp3+ and IFN-γ−Foxp3+ Treg cells (data are representative of three experiments). Author information * Abstract * Author information * Supplementary information Primary authors * These authors contributed equally to this work. * Clare M Baecher-Allan & * David A Hafler Affiliations * Departments of Neurology and Immunobiology, Yale School of Medicine, New Haven, Connecticut, USA. * Margarita Dominguez-Villar & * David A Hafler * Harvard Medical School, Boston, Massachusetts, USA. * Margarita Dominguez-Villar, * Clare M Baecher-Allan & * David A Hafler Contributions M.D.-V. designed and performed the experiments, analyzed data and wrote the manuscript; C.M.B.-A. and D.A.H. supervised the study and wrote the manuscript. Competing financial interests The authors declare no competing financial interests. Corresponding author Correspondence to: * David A Hafler Author Details * Margarita Dominguez-Villar Search for this author in: * NPG journals * PubMed * Google Scholar * Clare M Baecher-Allan Search for this author in: * NPG journals * PubMed * Google Scholar * David A Hafler Contact David A Hafler Search for this author in: * NPG journals * PubMed * Google Scholar Supplementary information * Abstract * Author information * Supplementary information PDF files * Supplementary Text and Figures (561K) Supplementary Figures 1–15, Supplementary Table 1 and Supplementary Methods Additional data
• Host-mediated regulation of superinfection in malaria
  - Nat Med 17(6):732-737 (2011)
  Nature Medicine | Brief Communication Identification of T helper type 1–like, Foxp3+ regulatory T cells in human autoimmune disease * Margarita Dominguez-Villar1, 2 * Clare M Baecher-Allan2, 3 * David A Hafler1, 2, 3 * Affiliations * Contributions * Corresponding authorJournal name:Nature MedicineVolume: 17,Pages:673–675Year published:(2011)DOI:doi:10.1038/nm.2389Received09 November 2010Accepted28 April 2010Published online03 May 2011 Abstract * Abstract * Author information * Supplementary information Article tools * Full text * 日本語要約 * Print * Email * Download PDF * Download citation * Order reprints * Rights and permissions * Share/bookmark * Connotea * CiteULike * Facebook * Twitter * Delicious * Digg CD4+CD25highCD127low/– forkhead box p3 (Foxp3)+ regulatory T cells (Treg cells) possess functional plasticity. Here we describe a higher frequency of T helper type 1 (TH1)-like, interferon-γ (IFN-γ)-secreting Foxp3+ T cells in untreated subjects with relapsing remitting multiple sclerosis (RRMS) as compared to healthy control individuals. In subjects treated with IFN-β, the frequency of IFN-γ+Foxp3+ T cells is similar to that in healthy control subjects. In vitro, human Treg cells from healthy subjects acquire a TH1-like phenotype when cultured in the presence of interleukin-12 (IL-12). TH1-like Treg cells show reduced suppressive activity in vitro, which can partially be reversed by IFN-γ–specific antibodies or by removal of IL-12. View full text Figures at a glance * Figure 1: Treg cells from individuals with RRMS secrete IFN-γex vivo. () The frequency of FACS-sorted IFN-γ+ and IL-17+ Treg cells in healthy control individuals (left) and untreated individuals with RRMS (middle, n = 17) gated on Foxp3+ Treg cells. Right, purity analysis of the sorted IFN-γ+Foxp3+ and IFN-γ−Foxp3+ populations from subjects with RRMS used for methylation analysis in . () Percentage of IFN-γ+Foxp3+ and IL-17+Foxp3+ Treg cells (n = 17) as a proportion of total Foxp3+ Treg cells. () Representative example of methylation analysis of the TSDR region of the FOXP3 locus in sorted IFN-γ+Foxp3+ and IFN-γ−Foxp3+ Treg cells from subjects with RRMS. An analysis of IFN-γ+Foxp3− memory T cells from subjects with RRMS is shown as a control. () Proliferation of responder T (Tresp) cells cultured with ex vivo FACS-sorted Treg cells from healthy control subjects and untreated subjects with multiple sclerosis (MS; Treg cell:Tresp cell ratio of 1:2) in the presence or absence of an IFN-γ–specific antibody (n = 4). () The frequency! of IFN-γ+ and IL-17+ Treg cells in healthy control subjects (left) or IFN-β–treated patients with RRMS (right) as assessed by intracellular cytokine staining and FACS analysis. The bar diagram (right) shows the percentage of IFN-γ+Foxp3+ and IL-17+Foxp3+ cells as a proportion of total Foxp3+ Treg cells in healthy controls or IFN-β–treated patients with RRMS (n = 12). Approval for studies was obtained from the Brigham and Women's Hospital Institutional Review Board, and informed consent was obtained from all donors. * Figure 2: Characterization of IL-12–driven, IFN-γ+Foxp3+ Treg cells in vitro from healthy controls. () Intracellular staining for IFN-γ, IL-4, IL-17 and IL-10 of untreated (upper row) and IL-12-stimulated (bottom row) human Treg cells from healthy controls at day 4. () mRNA expression of FOXP3 (left), GATA3 (middle) and TBX21 (right) in Treg cells stimulated in the presence or absence of IL-12 for 5 d (data are a representative example of three experiments performed with similar results; *P < 0.05). () Proliferation of Tresp cells cocultured for 3 d with Treg cells (Treg cell:Tresp cell ratio of 1:2) previously treated with IL-2 (top) or IL-2 + IL-12 (bottom), as assessed by carboxyfluorescein succinimidyl ester (CFSE) dilution. Histograms depict unstimulated Tresp cells alone (top left), Tresp cells stimulated with antibody to CD3 (anti-CD3) and antigen-presenting cells without Treg cells (bottom left) and Tresp cell and Treg cell cocultures without blocking antibodies (second column), in the presence of an IL-10–specific blocking antibody (anti–IL-10; third column),! IFN-γ–specific blocking antibody (anti–IFN-γ; fourth column) or both antibodies (fifth column). () Representative example of staining for TH1-associated chemokines (CCR5 and CXCR3) and TGF-β on IFN-γ+Foxp3+ and IFN-γ−Foxp3+ Treg cells (data are representative of three experiments). Author information * Abstract * Author information * Supplementary information Primary authors * These authors contributed equally to this work. * Clare M Baecher-Allan & * David A Hafler Affiliations * Departments of Neurology and Immunobiology, Yale School of Medicine, New Haven, Connecticut, USA. * Margarita Dominguez-Villar & * David A Hafler * Harvard Medical School, Boston, Massachusetts, USA. * Margarita Dominguez-Villar, * Clare M Baecher-Allan & * David A Hafler Contributions M.D.-V. designed and performed the experiments, analyzed data and wrote the manuscript; C.M.B.-A. and D.A.H. supervised the study and wrote the manuscript. Competing financial interests The authors declare no competing financial interests. Corresponding author Correspondence to: * David A Hafler Author Details * Margarita Dominguez-Villar Search for this author in: * NPG journals * PubMed * Google Scholar * Clare M Baecher-Allan Search for this author in: * NPG journals * PubMed * Google Scholar * David A Hafler Contact David A Hafler Search for this author in: * NPG journals * PubMed * Google Scholar Supplementary information * Abstract * Author information * Supplementary information PDF files * Supplementary Text and Figures (561K) Supplementary Figures 1–15, Supplementary Table 1 and Supplementary Methods Additional data
• A clinical microchip for evaluation of single immune cells reveals high functional heterogeneity in phenotypically similar T cells
  - Nat Med 17(6):738-743 (2011)
  Nature Medicine | Brief Communication Identification of T helper type 1–like, Foxp3+ regulatory T cells in human autoimmune disease * Margarita Dominguez-Villar1, 2 * Clare M Baecher-Allan2, 3 * David A Hafler1, 2, 3 * Affiliations * Contributions * Corresponding authorJournal name:Nature MedicineVolume: 17,Pages:673–675Year published:(2011)DOI:doi:10.1038/nm.2389Received09 November 2010Accepted28 April 2010Published online03 May 2011 Abstract * Abstract * Author information * Supplementary information Article tools * Full text * 日本語要約 * Print * Email * Download PDF * Download citation * Order reprints * Rights and permissions * Share/bookmark * Connotea * CiteULike * Facebook * Twitter * Delicious * Digg CD4+CD25highCD127low/– forkhead box p3 (Foxp3)+ regulatory T cells (Treg cells) possess functional plasticity. Here we describe a higher frequency of T helper type 1 (TH1)-like, interferon-γ (IFN-γ)-secreting Foxp3+ T cells in untreated subjects with relapsing remitting multiple sclerosis (RRMS) as compared to healthy control individuals. In subjects treated with IFN-β, the frequency of IFN-γ+Foxp3+ T cells is similar to that in healthy control subjects. In vitro, human Treg cells from healthy subjects acquire a TH1-like phenotype when cultured in the presence of interleukin-12 (IL-12). TH1-like Treg cells show reduced suppressive activity in vitro, which can partially be reversed by IFN-γ–specific antibodies or by removal of IL-12. View full text Figures at a glance * Figure 1: Treg cells from individuals with RRMS secrete IFN-γex vivo. () The frequency of FACS-sorted IFN-γ+ and IL-17+ Treg cells in healthy control individuals (left) and untreated individuals with RRMS (middle, n = 17) gated on Foxp3+ Treg cells. Right, purity analysis of the sorted IFN-γ+Foxp3+ and IFN-γ−Foxp3+ populations from subjects with RRMS used for methylation analysis in . () Percentage of IFN-γ+Foxp3+ and IL-17+Foxp3+ Treg cells (n = 17) as a proportion of total Foxp3+ Treg cells. () Representative example of methylation analysis of the TSDR region of the FOXP3 locus in sorted IFN-γ+Foxp3+ and IFN-γ−Foxp3+ Treg cells from subjects with RRMS. An analysis of IFN-γ+Foxp3− memory T cells from subjects with RRMS is shown as a control. () Proliferation of responder T (Tresp) cells cultured with ex vivo FACS-sorted Treg cells from healthy control subjects and untreated subjects with multiple sclerosis (MS; Treg cell:Tresp cell ratio of 1:2) in the presence or absence of an IFN-γ–specific antibody (n = 4). () The frequency! of IFN-γ+ and IL-17+ Treg cells in healthy control subjects (left) or IFN-β–treated patients with RRMS (right) as assessed by intracellular cytokine staining and FACS analysis. The bar diagram (right) shows the percentage of IFN-γ+Foxp3+ and IL-17+Foxp3+ cells as a proportion of total Foxp3+ Treg cells in healthy controls or IFN-β–treated patients with RRMS (n = 12). Approval for studies was obtained from the Brigham and Women's Hospital Institutional Review Board, and informed consent was obtained from all donors. * Figure 2: Characterization of IL-12–driven, IFN-γ+Foxp3+ Treg cells in vitro from healthy controls. () Intracellular staining for IFN-γ, IL-4, IL-17 and IL-10 of untreated (upper row) and IL-12-stimulated (bottom row) human Treg cells from healthy controls at day 4. () mRNA expression of FOXP3 (left), GATA3 (middle) and TBX21 (right) in Treg cells stimulated in the presence or absence of IL-12 for 5 d (data are a representative example of three experiments performed with similar results; *P < 0.05). () Proliferation of Tresp cells cocultured for 3 d with Treg cells (Treg cell:Tresp cell ratio of 1:2) previously treated with IL-2 (top) or IL-2 + IL-12 (bottom), as assessed by carboxyfluorescein succinimidyl ester (CFSE) dilution. Histograms depict unstimulated Tresp cells alone (top left), Tresp cells stimulated with antibody to CD3 (anti-CD3) and antigen-presenting cells without Treg cells (bottom left) and Tresp cell and Treg cell cocultures without blocking antibodies (second column), in the presence of an IL-10–specific blocking antibody (anti–IL-10; third column),! IFN-γ–specific blocking antibody (anti–IFN-γ; fourth column) or both antibodies (fifth column). () Representative example of staining for TH1-associated chemokines (CCR5 and CXCR3) and TGF-β on IFN-γ+Foxp3+ and IFN-γ−Foxp3+ Treg cells (data are representative of three experiments). Author information * Abstract * Author information * Supplementary information Primary authors * These authors contributed equally to this work. * Clare M Baecher-Allan & * David A Hafler Affiliations * Departments of Neurology and Immunobiology, Yale School of Medicine, New Haven, Connecticut, USA. * Margarita Dominguez-Villar & * David A Hafler * Harvard Medical School, Boston, Massachusetts, USA. * Margarita Dominguez-Villar, * Clare M Baecher-Allan & * David A Hafler Contributions M.D.-V. designed and performed the experiments, analyzed data and wrote the manuscript; C.M.B.-A. and D.A.H. supervised the study and wrote the manuscript. Competing financial interests The authors declare no competing financial interests. Corresponding author Correspondence to: * David A Hafler Author Details * Margarita Dominguez-Villar Search for this author in: * NPG journals * PubMed * Google Scholar * Clare M Baecher-Allan Search for this author in: * NPG journals * PubMed * Google Scholar * David A Hafler Contact David A Hafler Search for this author in: * NPG journals * PubMed * Google Scholar Supplementary information * Abstract * Author information * Supplementary information PDF files * Supplementary Text and Figures (561K) Supplementary Figures 1–15, Supplementary Table 1 and Supplementary Methods Additional data
• Visualizing the innate and adaptive immune responses underlying allograft rejection by two-photon microscopy
  - Nat Med 17(6):744-749 (2011)
  Nature Medicine | Brief Communication Identification of T helper type 1–like, Foxp3+ regulatory T cells in human autoimmune disease * Margarita Dominguez-Villar1, 2 * Clare M Baecher-Allan2, 3 * David A Hafler1, 2, 3 * Affiliations * Contributions * Corresponding authorJournal name:Nature MedicineVolume: 17,Pages:673–675Year published:(2011)DOI:doi:10.1038/nm.2389Received09 November 2010Accepted28 April 2010Published online03 May 2011 Abstract * Abstract * Author information * Supplementary information Article tools * Full text * 日本語要約 * Print * Email * Download PDF * Download citation * Order reprints * Rights and permissions * Share/bookmark * Connotea * CiteULike * Facebook * Twitter * Delicious * Digg CD4+CD25highCD127low/– forkhead box p3 (Foxp3)+ regulatory T cells (Treg cells) possess functional plasticity. Here we describe a higher frequency of T helper type 1 (TH1)-like, interferon-γ (IFN-γ)-secreting Foxp3+ T cells in untreated subjects with relapsing remitting multiple sclerosis (RRMS) as compared to healthy control individuals. In subjects treated with IFN-β, the frequency of IFN-γ+Foxp3+ T cells is similar to that in healthy control subjects. In vitro, human Treg cells from healthy subjects acquire a TH1-like phenotype when cultured in the presence of interleukin-12 (IL-12). TH1-like Treg cells show reduced suppressive activity in vitro, which can partially be reversed by IFN-γ–specific antibodies or by removal of IL-12. View full text Figures at a glance * Figure 1: Treg cells from individuals with RRMS secrete IFN-γex vivo. () The frequency of FACS-sorted IFN-γ+ and IL-17+ Treg cells in healthy control individuals (left) and untreated individuals with RRMS (middle, n = 17) gated on Foxp3+ Treg cells. Right, purity analysis of the sorted IFN-γ+Foxp3+ and IFN-γ−Foxp3+ populations from subjects with RRMS used for methylation analysis in . () Percentage of IFN-γ+Foxp3+ and IL-17+Foxp3+ Treg cells (n = 17) as a proportion of total Foxp3+ Treg cells. () Representative example of methylation analysis of the TSDR region of the FOXP3 locus in sorted IFN-γ+Foxp3+ and IFN-γ−Foxp3+ Treg cells from subjects with RRMS. An analysis of IFN-γ+Foxp3− memory T cells from subjects with RRMS is shown as a control. () Proliferation of responder T (Tresp) cells cultured with ex vivo FACS-sorted Treg cells from healthy control subjects and untreated subjects with multiple sclerosis (MS; Treg cell:Tresp cell ratio of 1:2) in the presence or absence of an IFN-γ–specific antibody (n = 4). () The frequency! of IFN-γ+ and IL-17+ Treg cells in healthy control subjects (left) or IFN-β–treated patients with RRMS (right) as assessed by intracellular cytokine staining and FACS analysis. The bar diagram (right) shows the percentage of IFN-γ+Foxp3+ and IL-17+Foxp3+ cells as a proportion of total Foxp3+ Treg cells in healthy controls or IFN-β–treated patients with RRMS (n = 12). Approval for studies was obtained from the Brigham and Women's Hospital Institutional Review Board, and informed consent was obtained from all donors. * Figure 2: Characterization of IL-12–driven, IFN-γ+Foxp3+ Treg cells in vitro from healthy controls. () Intracellular staining for IFN-γ, IL-4, IL-17 and IL-10 of untreated (upper row) and IL-12-stimulated (bottom row) human Treg cells from healthy controls at day 4. () mRNA expression of FOXP3 (left), GATA3 (middle) and TBX21 (right) in Treg cells stimulated in the presence or absence of IL-12 for 5 d (data are a representative example of three experiments performed with similar results; *P < 0.05). () Proliferation of Tresp cells cocultured for 3 d with Treg cells (Treg cell:Tresp cell ratio of 1:2) previously treated with IL-2 (top) or IL-2 + IL-12 (bottom), as assessed by carboxyfluorescein succinimidyl ester (CFSE) dilution. Histograms depict unstimulated Tresp cells alone (top left), Tresp cells stimulated with antibody to CD3 (anti-CD3) and antigen-presenting cells without Treg cells (bottom left) and Tresp cell and Treg cell cocultures without blocking antibodies (second column), in the presence of an IL-10–specific blocking antibody (anti–IL-10; third column),! IFN-γ–specific blocking antibody (anti–IFN-γ; fourth column) or both antibodies (fifth column). () Representative example of staining for TH1-associated chemokines (CCR5 and CXCR3) and TGF-β on IFN-γ+Foxp3+ and IFN-γ−Foxp3+ Treg cells (data are representative of three experiments). Author information * Abstract * Author information * Supplementary information Primary authors * These authors contributed equally to this work. * Clare M Baecher-Allan & * David A Hafler Affiliations * Departments of Neurology and Immunobiology, Yale School of Medicine, New Haven, Connecticut, USA. * Margarita Dominguez-Villar & * David A Hafler * Harvard Medical School, Boston, Massachusetts, USA. * Margarita Dominguez-Villar, * Clare M Baecher-Allan & * David A Hafler Contributions M.D.-V. designed and performed the experiments, analyzed data and wrote the manuscript; C.M.B.-A. and D.A.H. supervised the study and wrote the manuscript. Competing financial interests The authors declare no competing financial interests. Corresponding author Correspondence to: * David A Hafler Author Details * Margarita Dominguez-Villar Search for this author in: * NPG journals * PubMed * Google Scholar * Clare M Baecher-Allan Search for this author in: * NPG journals * PubMed * Google Scholar * David A Hafler Contact David A Hafler Search for this author in: * NPG journals * PubMed * Google Scholar Supplementary information * Abstract * Author information * Supplementary information PDF files * Supplementary Text and Figures (561K) Supplementary Figures 1–15, Supplementary Table 1 and Supplementary Methods Additional data