mimmunМедицинская иммунологияMedical Immunology (Russia)1563-06252313-741XSPb RAACI10.15789/1563-0625-IVM-1975mimmun-1975Research ArticleОРИГИНАЛЬНЫЕ СТАТЬИORIGINAL ARTICLESПоддержание CD4+ центральных и эффекторных клеток памяти в норме и в модели воспаления in vitroIn vitro maintaining of CD4+ central and effector memory cells in normal and inflammatory conditionshttps://orcid.org/0000-0003-3327-3630БлиноваЕ. А.BlinovaE. A.

Блинова Елена Андреевна – кандидат биологических наук, старший научный сотрудник лаборатории клинической иммунопатологии

630099, г. Новосибирск, ул. Ядринцевская, 14

Blinova Elena A. , PhD (Biology), Senior Research Associate, Laboratory of Clinical Immunopathology

630099, Novosibirsk, Yadrintsevskaya str., 14.

blinovaelena-85@yandex.ruКолероваА. В.KolerovaA. V.

Колерова Анастасия Викторовна – аспирант лаборатории клинической иммунопатологии

Новосибирск

Kolerova Anstasiia V., Postgraduate Student, Laboratory of Clinical Immunopathology

Novosibirsk

periosteum@mail.ruБалясниковВ. Е.BalyasnikovV. E.

Балясников Владимир Евгеньевич – студент лечебного факультета

Новосибирск

Balyasnikov Vladimir E., Student

Novosibirsk

vladimirtatarsk@gmail.comКозловВ. А.KozlovV. A.

Козлов Владимир Александрович – доктор медицинских наук, профессор, академик РАН, заведующий лабораторией клинической иммунопатологии, научный консультант

Новосибирск

Kozlov Vladimir A., PhD, MD (Medicine), Professor, Full Member, Russian Academy of Sciences, Head, Laboratory of Clinical Immunopathology, Scientific Advisor

Novosibirsk

vakoz40@yndex.ruФГБНУ «Научно-исследовательский институт клинической и фундаментальной иммунологии»РоссияResearch Institute of Fundamental and Clinical ImmunologyRussian FederationФГБОУ ВО «Новосибирский государственный медицинский университет»РоссияNovosibirsk State Medical UniversityRussian Federation202027112020225837846Copyright © Блинова Е.А., Колерова А.В., Балясников В.Е., Козлов В.А., 20202020Блинова Е.А., Колерова А.В., Балясников В.Е., Козлов В.А.Blinova E.A., Kolerova A.V., Balyasnikov V.E., Kozlov V.A.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.mimmun.ru/mimmun/article/view/1975

В организме ключевым фактором для выживания и поддержания CD4+ центральных (Tcm) и эффекторных (Tem) клеток памяти является IL-7. При многих аутоиммунных заболеваниях детектируется повышенный уровень IL-7 в сыворотке крови, очаге воспаления, что ставит вопрос об участии данного гомеостатического фактора в выживании Т-клеток памяти, в том числе аутореактивных, в условиях воспаления. Целью исследования явилось изучение механизмов поддержания CD4+Т-клеток памяти в норме и в условиях воспаления. Мы разработали модель воспаления in vitro, основанную на индукции провоспалительных цитокинов, и оценили влияние IL-7 на сортированные популяции CD4+Tcm и Tem в норме и в модели воспаления in vitro. IL-7 способствовал сохранению CD4+Tcm во всех вариантах культур. В отсутствие контакта с прилипающей фракцией пролиферативный ответ Tcm и Tem на IL-7 был немного снижен как в норме, так и в условиях воспаления. Что говорит о меньшей чувствительности Т-клеток памяти к контактам с MHC и, возможно, необходимости в дополнительных сигналах для полноценной стимуляции IL-7. В пользу данного утверждения свидетельствуют и данные об экспрессии CD127 и CD132: в отсутствии контакта с MHC доля CD127+CD132+ клеток уменьшалась в обеих субпопуляциях CD4+ клеток памяти. В культурах in vitro IL-7 способствовал снижению экспрессии CD127 и увеличению экспрессии CD132 на CD4+Tcm и Tem. Мы охарактеризовали CD4+Tcm и Tem по аффинности Т-клетокчного рецептора (ТКР), используя уровень экспрессии CD5. Известно, что клетки с высокой аффинностью ТКР к собственным антигенам обладают более высокой экспрессией CD5. Tcm содержали большее количество CD5high клеток по сравнению с Tem. В культурах IL-7 способствовал поддержанию высокого уровня экспрессии CD5 на Tcm, который был сопоставим с уровнем экспрессии в периферической крови. Высокая экспрессия CD5 на Tem наблюдалась при добавлении IL-7 в модели воспаления in vitro. В отсутствие контакта с MHC число CD5high среди CD4+Tem и Tcm снижалось. Таким образом, CD4+Tcm клетки с высоким родством к аутологичным антигенам, возможно, зависимы от наличия гомеостатических факторов, в частности IL-7, и контакта с антигенпрезентирующими клетками (АПК). В условиях воспаления не было выявлено изменений в механизме поддержания CD4+Tcm, в отличие от CD4+Tem. Менее зависимые от IL-7 в обычных условиях, CD4+CD5highTem клетки накапливались в условиях воспаления в присутствии IL-7.

IL-7 is a key factor for the survival and maintenance of CD4+ central (Tcm) and effector (Tem) memory cells in the whole body. In many autoimmune diseases, an elevated level of IL-7 is detected in blood serum and at the site of inflammation, thus suggesting participation of this homeostatic factor in the survival of memory T cells, including auto-reactive clones, in inflammatory disorders. The aim of the study was to investigate the mechanisms of maintaining CD4+ memory T cells under normal and inflammatory conditions. We developed an in vitro model of inflammation, based on induction of pro-inflammatory cytokines, and then evaluated the effects of IL-7 upon purified sorted populations of CD4+Tcm and Tem under normal conditions and in vitro inflammatory model. IL-7 treatment promoted maintenance of CD4+Tcm phenotype in all variants of cultures. In the absence of contact with adherent cell fraction, the IL-7-induced proliferation of Tcm and Tem was slightly reduced, both under normal and inflammatory conditions, thus suggesting low sensitivity of memory T cells to contacts with MHC, and, probably, a requirement for additional signals to provide complete stimulation with IL-7. The last suggestion is also supported by data about CD127 and CD132 expression, i.e., in the absence of contact with MHC, the proportion of CD127+CD132+ cells was decreased in both subpopulations of CD4+ memory cells. Upon in vitro cultures, IL-7 contributed to decreased expression of CD127, and increased expression of CD132 on CD4+Tcm and Tem. We have evaluated the CD4+Tcm and Tem populations by affinity of T cell receptor (TCR), using the level of CD5 expression. Т cells with high TCR affinity for self-antigens are known to have higher expression of CD5. In comparison to Tem, the Tcm contained more CD5high cells. In cultures, IL-7 promoted a high level of CD5 expression on Tcm, which was comparable to levels observed in peripheral blood cells. High CD5 expression on Tem was observed after stimulation with IL-7 in the in vitro inflammatory model. In the absence of contact with MHC, the number of CD5high cells decreased among CD4+Tem and Tcm. Thus, CD4+Tcm cells with high affinity for autologous antigens are probably dependent on the presence of homeostatic factors, in particular, IL-7, and contacts with antigen-presenting cells (APCs). Under conditions of inflammation, no changes were revealed in the mechanism of maintaining CD4+Tcm, in contrast to CD4+Tem. Being less dependent on IL-7 under normal conditions, CD4+CD5highTem are accumulated in the presence of IL-7 under in vitro inflammatory conditions.

центральные Т-клетки памятиэффекторные Т-клетки памятиIL-7рецептор для IL-7CD5провоспалительные цитокинымодель воспаления in vitrocentral memory T cellseffector memory T cellsIL-7IL-7 receptorCD5proinflammatory cytokinesinflammation model in vitroРоссийский научный фонд (проект №18-75-00068)RSF (project №18-75-00068)ReferencesДемьянов А.В., Котов А.Ю., Симбирцев А.С. Диагностическая ценность исследования уровней цитокинов в клинической практике // Цитокины и воспаление, 2003. Т. 2, № 3. С. 20-35.Demyanov A.V., Kotov A.Yu., Simbirtsev A.S. Diagnostic value of cytokine studies in clinical practice. Tsitokiny i vospalenie = Cytokines and Inflammation, 2003, Vol. 2, no. 3, pp. 20-35. (In Russ.)Azzam H.S., Grinberg A., Lui K., Shen H., Shore E.W., Love P.E. CD5 expression is developmentally regulated by T cell receptor (TCR) signals and TCR avidity. J. Exp. Med., 1998, Vol. 188, no. 12, pp. 2301-2311.Azzam H.S., Grinberg A., Lui K., Shen H., Shore E.W., Love P.E. CD5 expression is developmentally regulated by T cell receptor (TCR) signals and TCR avidity. J. Exp. Med., 1998, Vol. 188, no. 12, pp. 2301-2311.Bani L., David D., Moreau J.L., Cayota A., Nakarai T., Ritz J., Thèzу J. Expression of the IL-2 receptor gamma subunit in resting human CD4 T lymphocytes: mRNA is constitutively transcribed and the protein stored as an intracellular component. Int. Immunol., 1997, Vol. 9, no. 4, pp. 573-580.Bani L., David D., Moreau J.L., Cayota A., Nakarai T., Ritz J., Thèzу J. Expression of the IL-2 receptor gamma subunit in resting human CD4 T lymphocytes: mRNA is constitutively transcribed and the protein stored as an intracellular component. Int. Immunol., 1997, Vol. 9, no. 4, pp. 573-580.Bikker A., Hack C.E., Lafeber F.P., van Roon J.A. Interleukin-7: a key Mediator in T cell-driven autoimmunity, inflammation, and tissue destruction. Curr. Pharm. Des., 2012, Vol. 18, no. 16, pp. 2347-2356.Bikker A., Hack C.E., Lafeber F.P., van Roon J.A. Interleukin-7: a key Mediator in T cell-driven autoimmunity, inflammation, and tissue destruction. Curr. Pharm. Des., 2012, Vol. 18, no. 16, pp. 2347-2356.Caserta S., Zamoyska R. Memories are made of this: synergy of T cell receptor and cytokine signals in CD4(+) central memory cell survival. Trends Immunol., 2007, Vol. 28, no. 6, pp. 245-248.Caserta S., Zamoyska R. Memories are made of this: synergy of T cell receptor and cytokine signals in CD4(+) central memory cell survival. Trends Immunol., 2007, Vol. 28, no. 6, pp. 245-248.Churchman S.M., Ponchel F. Interleukin-7 in rheumatoid arthritis. Rheumatology (Oxford), 2008, Vol. 47, no. 6, pp. 753-759.Churchman S.M., Ponchel F. Interleukin-7 in rheumatoid arthritis. Rheumatology (Oxford), 2008, Vol. 47, no. 6, pp. 753-759.Friedlein G., El Hage F., Vergnon I., Richon C., Saulnier P., Lecluse Y., Caignard A., Boumsell L., Bismuth G., Chouaib S., Mami-Chouaib F. Human CD5 protects circulating tumor antigenspecific CTL from tumor-mediated activation-induced cell death. J. Immunol., 2007, Vol. 178, no. 11, pp. 6821-6827.Friedlein G., El Hage F., Vergnon I., Richon C., Saulnier P., Lecluse Y., Caignard A., Boumsell L., Bismuth G., Chouaib S., Mami-Chouaib F. Human CD5 protects circulating tumor antigenspecific CTL from tumor-mediated activation-induced cell death. J. Immunol., 2007, Vol. 178, no. 11, pp. 6821-6827.Fry T.J., Connick E., Fallon J., Lederman M.M., Liewehr D.J., Spritzler J., Steinberg S.M., Wood L.V., Yarchoan R., Zuckerman J., Landay A., Mackall C.L. A potential role for interleukin-7 in T-cell homeostasis. Blood, 2001, Vol. 97, no. 10, pp. 2983-2990.Fry T.J., Connick E., Fallon J., Lederman M.M., Liewehr D.J., Spritzler J., Steinberg S.M., Wood L.V., Yarchoan R., Zuckerman J., Landay A., Mackall C.L. A potential role for interleukin-7 in T-cell homeostasis. Blood, 2001, Vol. 97, no. 10, pp. 2983-2990.Gattinoni L., Speiser D.E., Lichterfeld M., Bonini C. T memory stem cells in health and disease. Nat. Med., 2017, Vol. 23, no. 1, pp. 18-27.Gattinoni L., Speiser D.E., Lichterfeld M., Bonini C. T memory stem cells in health and disease. Nat. Med., 2017, Vol. 23, no. 1, pp. 18-27.Henriques C.M., Rino J., Nibbs R.J., Graham G.J., Barata J.T. IL-7 induces rapid clathrin-mediated internalization and JAK3-dependent degradation of IL-7Ralpha in T cells. Blood, 2010, Vol. 115, no. 16, pp. 3269-3277.Henriques C.M., Rino J., Nibbs R.J., Graham G.J., Barata J.T. IL-7 induces rapid clathrin-mediated internalization and JAK3-dependent degradation of IL-7Ralpha in T cells. Blood, 2010, Vol. 115, no. 16, pp. 3269-3277.Hong C., Luckey M.A., Ligons D.L., Waickman A.T., Park J.Y., Kim G.Y., Keller H.R., Etzensperger R., Tai X., Lazarevic V., Feigenbaum L., Catalfamo M., Walsh S.T.R., Park J.-H. Activated T cells secrete an alternatively spliced form of common gamma-chain that inhibits cytokine signaling and exacerbates inflammation. Immunity, 2014, Vol. 40, no. 6, pp. 910-923.Hong C., Luckey M.A., Ligons D.L., Waickman A.T., Park J.Y., Kim G.Y., Keller H.R., Etzensperger R., Tai X., Lazarevic V., Feigenbaum L., Catalfamo M., Walsh S.T.R., Park J.-H. Activated T cells secrete an alternatively spliced form of common gamma-chain that inhibits cytokine signaling and exacerbates inflammation. Immunity, 2014, Vol. 40, no. 6, pp. 910-923.Lenz D.C., Kurz S.K., Lemmens E., Schoenberger S.P., Sprent J., Oldstone M.B., Homann D. IL-7 regulates basal homeostatic proliferation of antiviral CD4+ T cell memory. Proc. Natl. Acad. Sci. USA, 2004, Vol. 101, no. 25, pp. 9357-9362.Lenz D.C., Kurz S.K., Lemmens E., Schoenberger S.P., Sprent J., Oldstone M.B., Homann D. IL-7 regulates basal homeostatic proliferation of antiviral CD4+ T cell memory. Proc. Natl. Acad. Sci. USA, 2004, Vol. 101, no. 25, pp. 9357-9362.Leung S., Liu X., Fang L., Chen X., Guo T., Zhang J. The cytokine milieu in the interplay of pathogenic Th1/ Th17 cells and regulatory T cells in autoimmune disease. Cell. Mol. Immunol., 2010, Vol. 7, no. 3, pp. 182-189.Leung S., Liu X., Fang L., Chen X., Guo T., Zhang J. The cytokine milieu in the interplay of pathogenic Th1/ Th17 cells and regulatory T cells in autoimmune disease. Cell. Mol. Immunol., 2010, Vol. 7, no. 3, pp. 182-189.Maldonado A., Mueller Y.M., Thomas P., Bojczuk P., O’Connors C., Katsikis P.D. Decreased effector memory CD45RA+CD62LCD8+ T cells and increased central memory CD45RA-CD62L+CD8+ T cells in peripheral blood of rheumatoid arthritis patients. Arthritis Res. Ther., 2003, Vol. 5, no. 2, pp. R91-R96.Maldonado A., Mueller Y.M., Thomas P., Bojczuk P., O’Connors C., Katsikis P.D. Decreased effector memory CD45RA+CD62LCD8+ T cells and increased central memory CD45RA-CD62L+CD8+ T cells in peripheral blood of rheumatoid arthritis patients. Arthritis Res. Ther., 2003, Vol. 5, no. 2, pp. R91-R96.Mazzucchelli R., Durum S.K. Interleukin-7 receptor expression: intelligent design. Nat. Rev. Immunol., 2007, Vol. 7, no. 2, pp. 144-154.Mazzucchelli R., Durum S.K. Interleukin-7 receptor expression: intelligent design. Nat. Rev. Immunol., 2007, Vol. 7, no. 2, pp. 144-154.Nakarai T., Robertson M.J., Streuli M., Wu Z., Ciardelli T.L., Smith K.A., Ritz J. Interleukin 2 receptor gamma chain expression on resting and activated lymphoid cells. J. Exp. Med., 1994, Vol. 180, no. 1, pp. 241-251.Nakarai T., Robertson M.J., Streuli M., Wu Z., Ciardelli T.L., Smith K.A., Ritz J. Interleukin 2 receptor gamma chain expression on resting and activated lymphoid cells. J. Exp. Med., 1994, Vol. 180, no. 1, pp. 241-251.Ordoñez-Rueda D., Lozano F., Sarukhan A., Raman C., Garcia-Zepeda E.A., Soldevila G. Increased numbers of thymic and peripheral CD4+CD25+Foxp3+ cells in the absence of CD5 signaling. Eur. J. Immunol., 2009, Vol. 39, no. 8, pp. 2233-2247.Ordoñez-Rueda D., Lozano F., Sarukhan A., Raman C., Garcia-Zepeda E.A., Soldevila G. Increased numbers of thymic and peripheral CD4+CD25+Foxp3+ cells in the absence of CD5 signaling. Eur. J. Immunol., 2009, Vol. 39, no. 8, pp. 2233-2247.Paukku K., Silvennoinen O. STATs as critical mediators of signal transduction and transcription: lessons learned from STAT5. Cytokine Growth Factor Rev., 2004, Vol. 15, no. 6, pp. 435-455.Paukku K., Silvennoinen O. STATs as critical mediators of signal transduction and transcription: lessons learned from STAT5. Cytokine Growth Factor Rev., 2004, Vol. 15, no. 6, pp. 435-455.Persaud S.P., Parker C.R., Lo W.L., Weber K.S., Allen P.M. Intrinsic CD4+ T cell sensitivity and response to a pathogen are set and sustained by avidity for thymic and peripheral complexes of self peptide and MHC. Nat. Immunol., 2014, Vol. 15, no. 3, pp. 266-274.Persaud S.P., Parker C.R., Lo W.L., Weber K.S., Allen P.M. Intrinsic CD4+ T cell sensitivity and response to a pathogen are set and sustained by avidity for thymic and peripheral complexes of self peptide and MHC. Nat. Immunol., 2014, Vol. 15, no. 3, pp. 266-274.Pickens S.R., Chamberlain N.D., Volin M.V. Pope R.M., Talarico N.E., Mandelin A.M. 2nd, Shahrara S. Characterization of interleukin-7 and interleukin-7 receptor in the pathogenesis of rheumatoid arthritis. Arthritis Rheum., 2011, Vol. 63, no. 10, pp. 2884-2893.Pickens S.R., Chamberlain N.D., Volin M.V. Pope R.M., Talarico N.E., Mandelin A.M. 2nd, Shahrara S. Characterization of interleukin-7 and interleukin-7 receptor in the pathogenesis of rheumatoid arthritis. Arthritis Rheum., 2011, Vol. 63, no. 10, pp. 2884-2893.Read K.A., Powell M.D., McDonald P.W., Oestreich K.J. IL-2, IL-7, and IL-15: multistage regulators of CD4(+) T helper cell differentiation. Exp. Hematol., 2016, Vol. 44, no. 9, pp. 799-808.Read K.A., Powell M.D., McDonald P.W., Oestreich K.J. IL-2, IL-7, and IL-15: multistage regulators of CD4(+) T helper cell differentiation. Exp. Hematol., 2016, Vol. 44, no. 9, pp. 799-808.Sasson S.C., Zaunders J.J., Zanetti G., King E.M., Merlin K.M., Smith D.E., Stanley K.K., Cooper D.A., Kelleher A.D. Increased plasma interleukin-7 level correlates with decreased CD127 and increased CD132 extracellular expression on T cell subsets in patients with HIV-1 infection. J. Infect. Dis., 2006, Vol. 193, no. 4, pp. 505-514.Sasson S.C., Zaunders J.J., Zanetti G., King E.M., Merlin K.M., Smith D.E., Stanley K.K., Cooper D.A., Kelleher A.D. Increased plasma interleukin-7 level correlates with decreased CD127 and increased CD132 extracellular expression on T cell subsets in patients with HIV-1 infection. J. Infect. Dis., 2006, Vol. 193, no. 4, pp. 505-514.Seyfarth J., Mütze N., Antony Cruz J., Kummer S., Reinauer C., Mayatepek E., Meissner T., Jacobsen M. CD4+ T-cells with high common γ chain expression and disturbed cytokine production are enriched in children with type-1 diabetes. Front. Immunol., 2019, Vol. 10, 820. doi: 10.3389/fimmu.2019.00820.Seyfarth J., Mütze N., Antony Cruz J., Kummer S., Reinauer C., Mayatepek E., Meissner T., Jacobsen M. CD4+ T-cells with high common γ chain expression and disturbed cytokine production are enriched in children with type-1 diabetes. Front. Immunol., 2019, Vol. 10, 820. doi: 10.3389/fimmu.2019.00820.Waickman A.T., Park J.Y., Park J.H. The common gamma-chain cytokine receptor: tricks-and-treats for T cells. Cell. Mol. Life Sci., 2016, Vol. 73, no. 2, pp. 253-269.Waickman A.T., Park J.Y., Park J.H. The common gamma-chain cytokine receptor: tricks-and-treats for T cells. Cell. Mol. Life Sci., 2016, Vol. 73, no. 2, pp. 253-269.Wang X.S., Li B.Z., Hu L.F., Wen P.F., Zhang M., Pan H.F., Ye D.Q. Perspectives of the relationship between IL-7 and autoimmune diseases. Clin. Rheumatol., 2013, Vol. 32, no. 12, pp. 1703-1709.Wang X.S., Li B.Z., Hu L.F., Wen P.F., Zhang M., Pan H.F., Ye D.Q. Perspectives of the relationship between IL-7 and autoimmune diseases. Clin. Rheumatol., 2013, Vol. 32, no. 12, pp. 1703-1709.Zhou X.Y., Yashiro-Ohtani Y., Toyo-Oka K., Park C.S., Tai X.G., Hamaoka T. and Fujiwara H. CD5 costimulation up-regulates the signaling to extracellular signal-regulated kinase activation in CD4+CD8+ thymocytes and supports their differentiation to the CD4 lineage. J. Immunol., 2000, Vol. 164, no. 3, pp. 1260-1268.Zhou X.Y., Yashiro-Ohtani Y., Toyo-Oka K., Park C.S., Tai X.G., Hamaoka T. and Fujiwara H. CD5 costimulation up-regulates the signaling to extracellular signal-regulated kinase activation in CD4+CD8+ thymocytes and supports their differentiation to the CD4 lineage. J. Immunol., 2000, Vol. 164, no. 3, pp. 1260-1268.

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