rspНаучно-практическая ревматологияRheumatology Science and Practice1995-44841995-4492IMA-PRESS, LLC10.14412/1995-4484-2016-660-666rsp-2319Research ArticleОРИГИНАЛЬНЫЕ ИССЛЕДОВАНИЯORIGINAL RESEARCHОсобенности фенотипа Т-регуляторных клеток при раннем ревматоидном артритеPHENOTYPIC FEATURES OF T REGULATORY CELLS IN EARLY RHEUMATOID ARTHRITISАвдееваА. С.AvdeevaA. S.

115522 Москва, Каширское шоссе, 34А

34A, Kashirskoe Shosse, Moscow 115522

9056249400@mail.ruРубцовЮ. П.RubtsovYu. P.

кафедра биохимии и молекулярной медицины факультета фундаментальной медицины

119192 Москва, Ломоносовский проспект, 31, корп. 5

Department of Biochemistry and Molecular Medicine, Faculty of Fundamental Medicine

31, Lomonosovsky Prospect, Build. 5, Moscow 119192

ПопковаТ. В.PopkovaT. V.

115522 Москва, Каширское шоссе, 34А

34A, Kashirskoe Shosse, Moscow 115522

ДыйкановД. Т.DyikanovD. T.

кафедра биохимии и молекулярной медицины факультета фундаментальной медицины

119192 Москва, Ломоносовский проспект, 31, корп. 5

Department of Biochemistry and Molecular Medicine, Faculty of Fundamental Medicine

31, Lomonosovsky Prospect, Build. 5, Moscow 119192

НасоновЕ. Л.NasonovE. L.

115522 Москва, Каширское шоссе, 34А

кафедра ревматологии Института профессионального образования 119991 Москва, ул. Трубецкая, 8, стр. 2

34A, Kashirskoe Shosse, Moscow 115522

Department of Rheumatology 8, Trubetskaya St., Build. 2, Moscow 119991

 

ФГБНУ «Научно-исследовательский институт ревматологии им. В.А. Насоновой»РоссияV.A. Nasonova Research Institute of RheumatologyRussian FederationФГБОУ ВО «Московский государственный университет им. М.В. Ломоносова»РоссияM.V. Lomonosov Moscow State UniversityRussian FederationФГБНУ «Научно-исследовательский институт ревматологии им. В.А. Насоновой» ФГБОУ ВО «Первый Московский государственный медицинский университет им. И.М. Сеченова» Минздрава РоссииРоссияV.A. Nasonova Research Institute of Rheumatology Institute of Professional Education, I.M. Sechenov First Moscow State Medical University, Ministry of Health of RussiaRussian Federation201617012017546660666Copyright © Авдеева А.С., Рубцов Ю.П., Попкова Т.В., Дыйканов Д.Т., Насонов Е.Л., 20172017Авдеева А.С., Рубцов Ю.П., Попкова Т.В., Дыйканов Д.Т., Насонов Е.Л.Avdeeva A.S., Rubtsov Y.P., Popkova T.V., Dyikanov D.T., Nasonov E.L.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://rsp.mediar-press.net/rsp/article/view/2319

Цель – изучить содержание и особенности фенотипа Т-регуляторных клеток (Трег) в периферической крови здоровых доноров и пациентов с ранним ревматоидным артритом (РА) методом многоцветной проточной цитометрии.

Материал и методы

Материал и методы. В исследование было включено 39 пациентов с ранним РА. Процентное количество (ПК) и абсолютное содержание Трег (FoxP3+CD25+; CD152+surface; CD152+intracellular; FoxP3+CD127; CD25+CD127-; FoxP3+ICOS+; FoxP3+CD154+; FoxP3+CD274+) определялось методом многоцветной проточной цитофлуориметрии. Контрольную группу составили 20 здоровых доноров, сопоставимых по полу и возрасту с обследованными пациентами.

Результаты и обсуждение

Результаты и обсуждение. Среди включенных в исследование больных медиана [25-й; 75-й перцентили] DAS28 составили 5,01 [4,2; 5,8], при этом у 22 (48,9%) регистрировалась высокая, у 20 (44,4%) – умеренная и у 3 (6,7%) – низкая активность патологического процесса. У пациентов с ранним РА по сравнению со здоровыми донорами регистрировались более низкое ПК FoxP3+CD25+ клеток, ПК и абсолютное содержание FoxP3+ICOS+, FoxP3+CD154+ и FoxP3+ CD274+ Т-клеток (p<0,05 во всех случаях). Регистрировалась отрицательная корреляционная взаимосвязь: ПК FoxP3+CD25+ с СРБ (r=-0,4); ПК CD152+intracellular с DAS28 (r=-0,35), СОЭ (r=-0,46) и СРБ (r=-0,54); ПК FoxP3+CD127- с СРБ (r=-0,42); ПК CD25+CD127- с DAS28 (r=-0,38), SDAI (r=-0,41), CDAI (r=-0,36), СОЭ (r=-0,39), СРБ (r=-0,47); р<0,05 во всех случаях.

Заключение

Заключение. Полученные данные свидетельствуют о нарушении функциональной активности Трег при раннем РА, что оказывает влияние на активность воспалительного процесса.

Objective

Objective: to investigate the count and characteristics of the phenotype of T regulatory cells (Treg) in the peripheral blood of healthy donors and patients with early rheumatoid arthritis (RA), by using multicolor flow cytometry.

Subjects and methods

Subjects and methods. The investigation enrolled 39 patients with early RA. The percentage and absolute count of Treg (FoxP3+CD25+, surface CD152+, intracellular CD152+, FoxP3+CD127, CD25+CD127, FoxP3+ICOS+, FoxP3+CD154+; and FoxP3+CD274+) was determined by multicolor flow-cytometry. A control group consisted of 20 healthy donors matched for sex and age with the examined patients.

Results and discussion

Results and discussion. In the patients included in the study, the median [25th; 75th percentiles] DAS28 was 5.01 [4.2; 5.8]; high, moderate, and low activity showed 22 (48.9%), 20 (44.4%), and 3 (6.7%) patients, respectively. The patients with early RA had a lower percentage of FoxP3+CD25+ cells and a lower percentage and absolute count of FoxP3+ICOS+, FoxP3+CD154+, and FoxP3+CD274+ T cells than the healthy donors (p<0.05 in all cases). There was a negative correlation of the percentage of FoxP3+CD25+ cells with C-reactive protein (CRP) (r = -0.4), that of intracellular CD152+ with DAS28 (r = -0.35), erythrocyte sedimentation rate (ESR) (r = -0.46), and CRP (r=-0.54); that of FoxP3+CD127 with CRP (r = -0.42); that of CD25+CD127 with DAS28 (r = -0.38), Simplified Disease Activity Index (r = -0.41), Clinical Disease Activity Index (r = -0.36), ESR (r = -0.39), and CRP (r = -0.47) (p < 0.05 in all cases).

Conclusion

Conclusion. The findings suggest that the functional activity of Treg is impaired in early RA, which has an impact on the activity of the inflammatory process.

ранний ревматоидный артритактивность заболеванияострофазовые показателиТ-регуляторные клетки.early rheumatoid arthritisdisease activityacute-phase proteinsT regulatory cellsReferencesНасонов ЕЛ, Каратеев ДЕ, Балабанова РМ. Ревматоидный артрит. В кн.: Насонов ЕЛ, Насонова ВА, редакторы. Ревматология: Национальное руководство. Москва: ГЭОТАР- Медиа; 2008. С. 290-331 [Nasonov EL, Karateev DE, Balabanova RM. Rheumatoid arthritis. In: Nasonov EL, Nasonova VA, editors. Revmatologiya: Natsional’noe rukovodstvo [Rheumatology: National guidelines]. Moscow: GEOTAR-Media; 2008. P. 290-331].Насонов ЕЛ, Каратеев ДЕ, Балабанова РМ. Ревматоидный артрит. В кн.: Насонов ЕЛ, Насонова ВА, редакторы. Ревматология: Национальное руководство. Москва: ГЭОТАР- Медиа; 2008. С. 290-331 [Nasonov EL, Karateev DE, Balabanova RM. Rheumatoid arthritis. In: Nasonov EL, Nasonova VA, editors. Revmatologiya: Natsional’noe rukovodstvo [Rheumatology: National guidelines]. Moscow: GEOTAR-Media; 2008. P. 290-331].Firestein G. Evolving concepts of rheumatoid arthritis. Nature. 2003;423:356-61. doi: 10.1038/nature01661Firestein G. Evolving concepts of rheumatoid arthritis. Nature. 2003;423:356-61. doi: 10.1038/nature01661Cope A. T cells in rheumatoid arthritis. Arthritis Res Ther. 2008;10 Suppl 1:S1. doi: 10.1186/ar2412Cope A. T cells in rheumatoid arthritis. Arthritis Res Ther. 2008;10 Suppl 1:S1. doi: 10.1186/ar2412Choy E. Selective modulation of T cell co-stimulation: a novel mode of action for the treatment of rheumatoid arthritis. Clin Exp Rheumatol. 2009;27:510-8.Choy E. Selective modulation of T cell co-stimulation: a novel mode of action for the treatment of rheumatoid arthritis. Clin Exp Rheumatol. 2009;27:510-8.Steward-Tharp S, Song Y, Siegel R, O’Shea J. New insights into T cells biology and T cells directed therapy for autoimmunity inflammation and immunosuppression. Ann NY Acad Sci. 2010;1183:123-48. doi: 10.1111/j.1749-6632.2009.05124.xSteward-Tharp S, Song Y, Siegel R, O’Shea J. New insights into T cells biology and T cells directed therapy for autoimmunity inflammation and immunosuppression. Ann NY Acad Sci. 2010;1183:123-48. doi: 10.1111/j.1749-6632.2009.05124.xБыковская СН, Насонов ЕЛ. Роль дефектов иммуносупрессии в развитии аутоиммунных заболеваний. Научно-практическая ревматология. 2005;(4):81-4 [Bykovskaya SN, Nasonov EL. Role of immunosupression defects in the development of autoimmune diseases. Nauchno- Prakticheskaya Revmatologiya = Rheumatology Science and Practiсе. 2005;43(4):81-4 (In Russ.)]. doi: 10.14412/1995-4484-2005-623Быковская СН, Насонов ЕЛ. Роль дефектов иммуносупрессии в развитии аутоиммунных заболеваний. Научно-практическая ревматология. 2005;(4):81-4 [Bykovskaya SN, Nasonov EL. Role of immunosupression defects in the development of autoimmune diseases. Nauchno- Prakticheskaya Revmatologiya = Rheumatology Science and Practiсе. 2005;43(4):81-4 (In Russ.)]. doi: 10.14412/1995-4484-2005-623Sakaguchi S, Yamaguchi T, Nomura T, Ono M. Regulatory T cells and immune tolerance. Cell. 2008;133(5):775-87. doi: 10.1016/j.cell.2008.05.009Sakaguchi S, Yamaguchi T, Nomura T, Ono M. Regulatory T cells and immune tolerance. Cell. 2008;133(5):775-87. doi: 10.1016/j.cell.2008.05.009Zeng H, Chi H. The interplay between regulatory T cells and metabolism in immune regulation. OncoImmunology. 2013;2(11):e26586. Epub 2013 Oct 21. doi: 10.4161/onci.26586Zeng H, Chi H. The interplay between regulatory T cells and metabolism in immune regulation. OncoImmunology. 2013;2(11):e26586. Epub 2013 Oct 21. doi: 10.4161/onci.26586Buckner JH. Mechanisms of impaired regulation by CD4+CD25+FOXP3+ regulatory T cells in human autoimmune diseases. Nat Rev Immunol. 2010;10:849-59. doi: 10.1038/nri2889Buckner JH. Mechanisms of impaired regulation by CD4+CD25+FOXP3+ regulatory T cells in human autoimmune diseases. Nat Rev Immunol. 2010;10:849-59. doi: 10.1038/nri2889Rudensky AY. Regulatory T cells and FoxP3. Immunol Rev. 2011;241;260-8. doi: 10.1111/j.1600-065X.2011.01018.xRudensky AY. Regulatory T cells and FoxP3. Immunol Rev. 2011;241;260-8. doi: 10.1111/j.1600-065X.2011.01018.xAbbas AK, Benoist C, Bluestone JA, et al. Regulatory T cells: recommendations to simplify the nomenclature. Nat Immunol. 2013;14:300-8. doi: 10.1038/ni.2554Abbas AK, Benoist C, Bluestone JA, et al. Regulatory T cells: recommendations to simplify the nomenclature. Nat Immunol. 2013;14:300-8. doi: 10.1038/ni.2554Miyara M, Yoshioka Y, Kitoh A, et al. Functional delineation and differentiation dynamics of human CD4+ T cells expressing the FoxP3 transcription factor. Immunity. 2009b;30:899- 911. doi: 10.1016/j.immuni.2009.03.019Miyara M, Yoshioka Y, Kitoh A, et al. Functional delineation and differentiation dynamics of human CD4+ T cells expressing the FoxP3 transcription factor. Immunity. 2009b;30:899- 911. doi: 10.1016/j.immuni.2009.03.019Miyara M, Ito Y, Sakaguchi S. T reg-cell therapies for autoimmune rheumatic duseases. Nat Rev Rheumatol. 2014 Sep;10(9):543-51. doi: 10.1038/nrhheum.2014.105Miyara M, Ito Y, Sakaguchi S. T reg-cell therapies for autoimmune rheumatic duseases. Nat Rev Rheumatol. 2014 Sep;10(9):543-51. doi: 10.1038/nrhheum.2014.105Prakken B, Wehrens E, van Wijl F. Quality or Quantity? Unraveling the role of T reg cells in rheumatoid arthritis. Arthritis Rheum. 2013;65:552-4. doi: 10.1002/art.37831Prakken B, Wehrens E, van Wijl F. Quality or Quantity? Unraveling the role of T reg cells in rheumatoid arthritis. Arthritis Rheum. 2013;65:552-4. doi: 10.1002/art.37831Klimiuk PA, Yang H, Goronzy JJ, Weyand CM. Production of cytokines and metalloproteinases in rheumatoid synovitis is T cell dependent. Clin Immunol. 1999;90:65- 78. doi: 10.1006/clim.1998.4618Klimiuk PA, Yang H, Goronzy JJ, Weyand CM. Production of cytokines and metalloproteinases in rheumatoid synovitis is T cell dependent. Clin Immunol. 1999;90:65- 78. doi: 10.1006/clim.1998.4618Lahl K, Loddenkemper C, Drouin C, et al. Selective depletion of Foxp3+ regulatory T cells induces a scurfy-like disease. J Exp Med. 2007;204(1):57-63. doi: 10.1084/jem.20061852. Epub 2007Jan 2.Lahl K, Loddenkemper C, Drouin C, et al. Selective depletion of Foxp3+ regulatory T cells induces a scurfy-like disease. J Exp Med. 2007;204(1):57-63. doi: 10.1084/jem.20061852. Epub 2007Jan 2.Cao D, Malmstrom V, Baecher-Allan C, et al. Isolation and functional characterization of regulatory CD25brightCD4+ T cells fromthe target organ of patients with rheumatoid arthritis. Eur J Immunol. 2003;33:215-23. doi: 10.1002/immu.200390024Cao D, Malmstrom V, Baecher-Allan C, et al. Isolation and functional characterization of regulatory CD25brightCD4+ T cells fromthe target organ of patients with rheumatoid arthritis. Eur J Immunol. 2003;33:215-23. doi: 10.1002/immu.200390024Cao D, van Vollenhoven R, Klareskog L, et al. CD25+CD4+ regulatory T cells are enriched in inflamed joints of patients with chronic rheumatic disease. Arthritis Res Ther. 2004;6:R335-46. doi: 10.1186/ar1192Cao D, van Vollenhoven R, Klareskog L, et al. CD25+CD4+ regulatory T cells are enriched in inflamed joints of patients with chronic rheumatic disease. Arthritis Res Ther. 2004;6:R335-46. doi: 10.1186/ar1192Van Amelsfort JMR, Jacobs KMG, Bijlsma JWJ, et al. CD4+CD25+ regulatory T cells in rheumatoid arthritis: differences in the presence, phenotype, and function between peripheral blood and synovial fluid. Arthritis Rheum. 2004;50:2775-85. doi: 10.1002/art.20499Van Amelsfort JMR, Jacobs KMG, Bijlsma JWJ, et al. CD4+CD25+ regulatory T cells in rheumatoid arthritis: differences in the presence, phenotype, and function between peripheral blood and synovial fluid. Arthritis Rheum. 2004;50:2775-85. doi: 10.1002/art.20499Mottonen M, Heikkinen J, Mustonen L, et al. CD4+ CD25+T cells with the phenotypic and functional characteristics of regulatory T cells are enriched in the synovial fluid of patients with rheumatoid arthritis. Clin Exper Immunol. 2005;140:360-7. doi: 10.1111/j.1365-2249.2005.02754.xMottonen M, Heikkinen J, Mustonen L, et al. CD4+ CD25+T cells with the phenotypic and functional characteristics of regulatory T cells are enriched in the synovial fluid of patients with rheumatoid arthritis. Clin Exper Immunol. 2005;140:360-7. doi: 10.1111/j.1365-2249.2005.02754.xLiu M-F, Wang C-R, Fung L-L, et al. The presence of cytokine-suppressive CD4+CD25+ T cells in the peripheral blood and synovial fluid of patients with rheumatoid arthritis. Scand J Immunol. 2005;62:312-7. doi: 10.1111/j.1365-3083.2005.01656.xLiu M-F, Wang C-R, Fung L-L, et al. The presence of cytokine-suppressive CD4+CD25+ T cells in the peripheral blood and synovial fluid of patients with rheumatoid arthritis. Scand J Immunol. 2005;62:312-7. doi: 10.1111/j.1365-3083.2005.01656.xCao D, Borjesson O, Larsson P, et al. FOXP3 identifies regulatory CD25brightCD4+ T cells in rheumatic joints. Scand J Immunol. 2006;63:444-52. doi: 10.1111/j.1365-3083.2006.001755.xCao D, Borjesson O, Larsson P, et al. FOXP3 identifies regulatory CD25brightCD4+ T cells in rheumatic joints. Scand J Immunol. 2006;63:444-52. doi: 10.1111/j.1365-3083.2006.001755.xJiao Z, Wang W, Jia R, et al. Accumulation of FoxP3-expressing CD4+CD25+ T cells with distinct chemokine receptors in synovial fluid of patients with active rheumatoid arthritis. Scand J Rheumatol. 2007;36:428-33. doi: 10.1080/03009740701482800Jiao Z, Wang W, Jia R, et al. Accumulation of FoxP3-expressing CD4+CD25+ T cells with distinct chemokine receptors in synovial fluid of patients with active rheumatoid arthritis. Scand J Rheumatol. 2007;36:428-33. doi: 10.1080/03009740701482800Moradi B, Schnatzer P, Hagmann S, et al. CD4+CD25+/highCD127low/- regulatory T cells are enriched in rheumatoid arthritis and osteoarthritis joints – analysis of frequency and phenotype in synovial membrane, synovial fluid and peripheral blood. Arthritis Res Ther. 2014;16: R97. doi: 10.1186/ar4545Moradi B, Schnatzer P, Hagmann S, et al. CD4+CD25+/highCD127low/- regulatory T cells are enriched in rheumatoid arthritis and osteoarthritis joints – analysis of frequency and phenotype in synovial membrane, synovial fluid and peripheral blood. Arthritis Res Ther. 2014;16: R97. doi: 10.1186/ar4545Dejaco C, Duftner C, Klauser A, Schirmer M. Altered T-cell subtypes in spondyloarthritis, rheumatoid arthritis and polymyalgia rheumatic. Rheumatol Int. 2010;30:297-303. doi: 10.1007/s00296-009-0949-9Dejaco C, Duftner C, Klauser A, Schirmer M. Altered T-cell subtypes in spondyloarthritis, rheumatoid arthritis and polymyalgia rheumatic. Rheumatol Int. 2010;30:297-303. doi: 10.1007/s00296-009-0949-9Sempere-Ortells JM, Perez-Garcia V, Martin-Alberca G, et al. Quantification and phenotype of regulatory T cells in rheumatoid arthritis according to disease activity Score- 28. Autoimmunity. 2009;42:636-45. doi: 10.3109/08916930903061491Sempere-Ortells JM, Perez-Garcia V, Martin-Alberca G, et al. Quantification and phenotype of regulatory T cells in rheumatoid arthritis according to disease activity Score- 28. Autoimmunity. 2009;42:636-45. doi: 10.3109/08916930903061491Kawashiri S-Y, Kawakami A, Okada A, et al. CD4+CD25(high)CD127(low/-) Treg cell frequency from peripheral blood correlates with disease activity in patients with rheumatoid arthritis. J Rheumatol. 2011;38:2517-21. doi: 10.3899/jrheum.110283Kawashiri S-Y, Kawakami A, Okada A, et al. CD4+CD25(high)CD127(low/-) Treg cell frequency from peripheral blood correlates with disease activity in patients with rheumatoid arthritis. J Rheumatol. 2011;38:2517-21. doi: 10.3899/jrheum.110283Han GM, O’Neil-Andersen NJ, Zurier RB, Lawrence DA. CD4+CD25high T cell numbers are enriched in the peripheral blood of patients with rheumatoid arthritis. Cell Immunol. 2008;253:92-101. doi: 10.1016/j.cellimm.2008.05.007Han GM, O’Neil-Andersen NJ, Zurier RB, Lawrence DA. CD4+CD25high T cell numbers are enriched in the peripheral blood of patients with rheumatoid arthritis. Cell Immunol. 2008;253:92-101. doi: 10.1016/j.cellimm.2008.05.007Lin SC, Chen K-H, Lin C-H, et al. The quantitative analysis of peripheral blood FOXP3- expressing T cells in systemic lupus erythematosus and rheumatoid arthritis patients. Eur J Clin Invest. 2007;37:987-96. doi: 10.1111/j.1365-2362.2007.01882.xLin SC, Chen K-H, Lin C-H, et al. The quantitative analysis of peripheral blood FOXP3- expressing T cells in systemic lupus erythematosus and rheumatoid arthritis patients. Eur J Clin Invest. 2007;37:987-96. doi: 10.1111/j.1365-2362.2007.01882.xJi L, Geng Y, Zhou W, Zhang Z. A study on relationship among apoptosis rates, number of peripheral T cell subtypes and disease activity in rheumatoid arthritis. Int J Rheum Dis. 2016;19:167-71. doi: 10.1111/1756-185X.12211Ji L, Geng Y, Zhou W, Zhang Z. A study on relationship among apoptosis rates, number of peripheral T cell subtypes and disease activity in rheumatoid arthritis. Int J Rheum Dis. 2016;19:167-71. doi: 10.1111/1756-185X.12211Dombrecht EJ, Aerts NE, Schuerwegh AJ, et al. Influence of antitumor necrosis factor therapy (Adalimumab) on regulatory T cells and dendritic cells in rheumatoid arthritis. Clin Exper Rheumatol. 2006;24:31-7.Dombrecht EJ, Aerts NE, Schuerwegh AJ, et al. Influence of antitumor necrosis factor therapy (Adalimumab) on regulatory T cells and dendritic cells in rheumatoid arthritis. Clin Exper Rheumatol. 2006;24:31-7.Lawson CA, Brown AK, Bejarano V, et al. Early rheumatoid arthritis is associated with a deficit in the CD4+CD25high regulatory T cell population in peripheral blood. Rheumatology (Oxford). 2006;45(10):1210-7. doi: 10.1093/rheumatology/kel089Lawson CA, Brown AK, Bejarano V, et al. Early rheumatoid arthritis is associated with a deficit in the CD4+CD25high regulatory T cell population in peripheral blood. Rheumatology (Oxford). 2006;45(10):1210-7. doi: 10.1093/rheumatology/kel089Hensor RMA, Hunt L, Patmar R, et al. Predicting the evaluation of inflammatory arthritis in ACPA-positive individuals: can T-cell subset help? Ann Rheum Dis. 2014;73 Suppl 1:A14. doi: 10.1136/annrheumdis-2013-205124.32Hensor RMA, Hunt L, Patmar R, et al. Predicting the evaluation of inflammatory arthritis in ACPA-positive individuals: can T-cell subset help? Ann Rheum Dis. 2014;73 Suppl 1:A14. doi: 10.1136/annrheumdis-2013-205124.32Linsley PS, Greene JL, Tan P, et al. Coexpression and functional cooperation of CTLA-4 and CD28 on activated T lymphocytes. J Exp Med. 1992;176:1595-604. doi: 10.1084/jem.176.6.1595Linsley PS, Greene JL, Tan P, et al. Coexpression and functional cooperation of CTLA-4 and CD28 on activated T lymphocytes. J Exp Med. 1992;176:1595-604. doi: 10.1084/jem.176.6.1595Harper K, Balzano C, Rouvier E, et al. CTLA-4 and CD28 activated lymphocyte molecules are closely related in both mouse and human as to sequence, message expression, gene structure, and chromosomal location. J Immunol. 1991;147:1037-44.Harper K, Balzano C, Rouvier E, et al. CTLA-4 and CD28 activated lymphocyte molecules are closely related in both mouse and human as to sequence, message expression, gene structure, and chromosomal location. J Immunol. 1991;147:1037-44.Yokosuka T, Kobayashi W, Takamatsu M, et al. Spatiotemporal basis of CTLA-4 costimulatory molecule-mediated negative regulation of T cell activation. Immunity. 2010;33:326-39. doi: 10.1016/j.immuni.2010.09.006Yokosuka T, Kobayashi W, Takamatsu M, et al. Spatiotemporal basis of CTLA-4 costimulatory molecule-mediated negative regulation of T cell activation. Immunity. 2010;33:326-39. doi: 10.1016/j.immuni.2010.09.006Cribbs AP, Kennedy A, Penn H, et al. Regulatory T cell function in rheumatoid arthritis is compromised by CTLA-4 promoter methylation resulting in a failure to activate the IDO pathway. Arthritis Rheum. 2014 Sep;66(9):2344-54. doi: 10.1002/art.38715Cribbs AP, Kennedy A, Penn H, et al. Regulatory T cell function in rheumatoid arthritis is compromised by CTLA-4 promoter methylation resulting in a failure to activate the IDO pathway. Arthritis Rheum. 2014 Sep;66(9):2344-54. doi: 10.1002/art.38715Schneider H, Downey J, Smith A, et al. Reversal of the TCR stop signal by CTLA-4. Science. 2006;313:1972-5. doi: 10.1126/science. 1131078Schneider H, Downey J, Smith A, et al. Reversal of the TCR stop signal by CTLA-4. Science. 2006;313:1972-5. doi: 10.1126/science. 1131078Hutloff A, Dittrich AM, Beier KC, et al. ICOS is an inducible Tcell co-stimulator structurally and functionally related to CD28. Nature. 1999;397:263-6. doi: 10.1038/16717Hutloff A, Dittrich AM, Beier KC, et al. ICOS is an inducible Tcell co-stimulator structurally and functionally related to CD28. Nature. 1999;397:263-6. doi: 10.1038/16717Yoshinaga SK, Whoriskey JS, Khare SD, et al. T-cell co-stimulation through B7RP-1 and ICOS. Nature. 1999;402:827-32. doi: 10.1038/45582Yoshinaga SK, Whoriskey JS, Khare SD, et al. T-cell co-stimulation through B7RP-1 and ICOS. Nature. 1999;402:827-32. doi: 10.1038/45582Kroczek RA, Mages HW, Hutloff A. Emerging paradigms of T-cell co-stimulation. Curr Opin Immunol. 2004;16:321-7. doi: 10.1016/j.coi.2004.03.002Kroczek RA, Mages HW, Hutloff A. Emerging paradigms of T-cell co-stimulation. Curr Opin Immunol. 2004;16:321-7. doi: 10.1016/j.coi.2004.03.002Greenwald RJ, Freeman GJ, Sharpe AH. The B7 family revisited. Ann Rev Immunol. 2005;23:515-48. doi: 10.1146/annurev.immunol.23.021704.115611Greenwald RJ, Freeman GJ, Sharpe AH. The B7 family revisited. Ann Rev Immunol. 2005;23:515-48. doi: 10.1146/annurev.immunol.23.021704.115611Sperling AI, Bluestone JA. ICOS costimulation: it’s not just for TH2 cells anymore. Nat Immunol. 2001;2:573-4. doi: 10.1038/89709Sperling AI, Bluestone JA. ICOS costimulation: it’s not just for TH2 cells anymore. Nat Immunol. 2001;2:573-4. doi: 10.1038/89709Bonhagen K, Liesenfeld O, Stadecker MJ, et al. ICOS Th cells produce distinct cytokines in different mucosal immune responses. Eur J Immunol. 2003;33:392-401. doi: 10.1002/immu.200310013Bonhagen K, Liesenfeld O, Stadecker MJ, et al. ICOS Th cells produce distinct cytokines in different mucosal immune responses. Eur J Immunol. 2003;33:392-401. doi: 10.1002/immu.200310013Burmeister Y, Lischke T, Dahler A, et al. ICOS controls the pool size of effector-memory and regulatory T cells. J Immunol. 2008;180:774-82. doi: 10.4049/jimmunol.180.2.774Burmeister Y, Lischke T, Dahler A, et al. ICOS controls the pool size of effector-memory and regulatory T cells. J Immunol. 2008;180:774-82. doi: 10.4049/jimmunol.180.2.774Hasegawa M, Fujimoto M, Matsushita T, et al. Augmented ICOS expression in patients with early diffuse cutaneous systemic sclerosis. Rheumatology. 2013;52:242-51. doi: 10.1093/rheumatology/kes258Hasegawa M, Fujimoto M, Matsushita T, et al. Augmented ICOS expression in patients with early diffuse cutaneous systemic sclerosis. Rheumatology. 2013;52:242-51. doi: 10.1093/rheumatology/kes258Grimbacher B, Hutloff A, Schlesier M, et al. Homozygous loss of ICOS is associated with adult-onset common variable immunodeficiency. Nat Immunol. 2003;4:261-8. doi: 10.1038/ni902Grimbacher B, Hutloff A, Schlesier M, et al. Homozygous loss of ICOS is associated with adult-onset common variable immunodeficiency. Nat Immunol. 2003;4:261-8. doi: 10.1038/ni902Bishop GA, Hostager BS. The CD40–CD154 interaction in B cell-T cell liaisons. Cytokine Growth Factor Rev. 2003;14:297-309. doi: 10.1016/S1359-6101(03)00024-8Bishop GA, Hostager BS. The CD40–CD154 interaction in B cell-T cell liaisons. Cytokine Growth Factor Rev. 2003;14:297-309. doi: 10.1016/S1359-6101(03)00024-8O’Sullivan B, Thomas R. CD40 and dendritic cell function. Crit Rev Immunol. 2003;23:83-107. doi: 10.1615/CritRevImmunol.v23.i12.50O’Sullivan B, Thomas R. CD40 and dendritic cell function. Crit Rev Immunol. 2003;23:83-107. doi: 10.1615/CritRevImmunol.v23.i12.50Peters A, Stunz L, Bishop G. CD40 and autoimmunity: the dark side of a great activator. Semin Immunol. 2009 Oct;21(5):293-300. doi: 10.1016/j.smim.2009.05.012Peters A, Stunz L, Bishop G. CD40 and autoimmunity: the dark side of a great activator. Semin Immunol. 2009 Oct;21(5):293-300. doi: 10.1016/j.smim.2009.05.012Hill D, Eastaff-Leung N, Bresatz-Atkins S, et al. Inhibition of activation induced CD154 on CD4+CD25- cells: a valid surrogate for human Treg suppressor function. Immunol Cell Biol. 2012;90: 812-21. doi: 10.1038/icb.2012.18Hill D, Eastaff-Leung N, Bresatz-Atkins S, et al. Inhibition of activation induced CD154 on CD4+CD25- cells: a valid surrogate for human Treg suppressor function. Immunol Cell Biol. 2012;90: 812-21. doi: 10.1038/icb.2012.18Sharpe AH, Wherry EJ, Ahmed R, Freeman GJ. The function of programmed cell death 1 and its ligands in regulating autoimmunity and infection. Nat Immunol. 2007;8:239-45. doi: 10.1038/ni1443Sharpe AH, Wherry EJ, Ahmed R, Freeman GJ. The function of programmed cell death 1 and its ligands in regulating autoimmunity and infection. Nat Immunol. 2007;8:239-45. doi: 10.1038/ni1443Okazaki T, Honjo T. PD-1 and PD-1 ligands: from discovery to clinical application. Int Immunol. 2007;19:813-24. doi: 10.1093/intimm/dxm057Okazaki T, Honjo T. PD-1 and PD-1 ligands: from discovery to clinical application. Int Immunol. 2007;19:813-24. doi: 10.1093/intimm/dxm057Keir ME, Butte MJ, Freeman GJ, Sharpe AH. PD-1 and its ligands in tolerance and immunity. Ann Rev Immunol. 2008;26:677-704. doi: 10.1146/annurev.immunol.26.021607.090331Keir ME, Butte MJ, Freeman GJ, Sharpe AH. PD-1 and its ligands in tolerance and immunity. Ann Rev Immunol. 2008;26:677-704. doi: 10.1146/annurev.immunol.26.021607.090331Francisco L, Sage P, Sharpe A. The PD-1 pathway in tolerance and autoimmunity. Immunol Rev. 2010;236:219-42. doi: 10.1111/j.1600-065X.2010.00923.xFrancisco L, Sage P, Sharpe A. The PD-1 pathway in tolerance and autoimmunity. Immunol Rev. 2010;236:219-42. doi: 10.1111/j.1600-065X.2010.00923.xFife B, Pauken K. The role of the PD-1 pathway in autoimmunity and peripheral tolerance. Ann NY Acad Sci. 2011;1217:45-59. doi: 10.1111/j.1749-6632.2010.05919.xFife B, Pauken K. The role of the PD-1 pathway in autoimmunity and peripheral tolerance. Ann NY Acad Sci. 2011;1217:45-59. doi: 10.1111/j.1749-6632.2010.05919.xBehrens F, Himsel A, Rehart S, et al. Imbalance in distribution of functional autologous regulatory T cells in rheumatoid arthritis. Ann Rheum Dis. 2007;66:1151-6. doi: 10.1136/ard.2006.068320Behrens F, Himsel A, Rehart S, et al. Imbalance in distribution of functional autologous regulatory T cells in rheumatoid arthritis. Ann Rheum Dis. 2007;66:1151-6. doi: 10.1136/ard.2006.068320

The authors declare that there are no conflicts of interest present.