nefrНефрологияNephrology (Saint-Petersburg)1561-62742541-9439Pavlov First Saint-Petersburg State Medical University10.24884/1561-6274-2012-16-4-45-49nefr-625Research ArticleОРИГИНАЛЬНЫЕ СТАТЬИ. КЛИНИЧЕСКИЕ ИССЛЕДОВАНИЯORIGINAL ARTICLES. CLINICAL INVESTIGATIONSЦИРКУЛИРУЮЩИЕ РЕГУЛЯТОРНЫЕ CD4+CD25+HIGHFOXP3+ Т-КЛЕТОК У РЕЦИПИЕНТОВ ПОЧЕЧНОГО ТРАНСПЛАНТАТА (ПРЕДВАРИТЕЛЬНОЕ СООБЩЕНИЕ)CIRCULATING REGULATIONAL T-CELLS CD4+CD25+HIGHFOXP3+ INKIDNEY TRANSPLANT RECIPIENTS (PRELIMINARY REPORT)КорюшкинаТ.KorushkinaT.

кафедра пропедевтики внутренних болезней

СиполА. В.SipolA. V.ДобронравовВ. А.DobronravovV. A.

кафедра пропедевтики внутренних болезней 

Добронравов В.А. 197022, Санкт-Петербург, ул. Л. Толстого, д. 17, тел/ факс: +7(812)234-66-56

dobronravov@nephrolog.ruНаучно-исследовательский институт нефрологии, Санкт-Петербургский государственный медицинский университет им.акад. И.П. ПавловаRussian FederationИнститут детской онкологии и гематологии им. Р.М. Горбачевой, СанктПетербургский государственный медицинский университет им. акад. И.П. ПавловаRussian Federation2012100420121644549Copyright © Корюшкина Т., Сипол А.В., Добронравов В.А., 20122012Корюшкина Т., Сипол А.В., Добронравов В.А.Korushkina T., Sipol A.V., Dobronravov V.A.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://journal.nephrolog.ru/jour/article/view/625

ЦЕЛЬ ИССЛЕДОВАНИЯ. Регуляторные Т-клетки (РТК) являются одним из механизмов развития и поддержания иммунной толерантности. Целью данного исследование была оценка содержания циркулирующих РТК, экспрессирующих маркеры CD4, CD25 и Foxp3, у больных с функционирующим почечным трансплантатом (ПТ). ПАЦИЕНТЫ И МЕТОДЫ. В обсервационное исследование были включены 30 стабильных реципиентов c ПТ со сроками посттрансплантационного наблюдения от 6 до 24 мес., в среднем 353 дня (95% ДИ 246–459), 7 больных через 1–3 мес. после трансплантации почки (ТП) и 5 здоровых лиц. Все больные получили индукцию анти-CD25 моноклональными антителами (анти-CD25МАb –базиликсимабом). Исследование экспрессии CD4, CD25 и FoxP3 в лимфоцитах периферической крови проводили методом проточной цитометрии. РЕЗУЛЬТАТЫ. Доля CD4+CD25 + high FOXP3+ от общего количества CD4+ клеток варьировала в достаточно широких пределах и была, в среднем, существенно ниже, чем у здоровых лиц. Общая экспрессия FoxP3 в лимфоцитах имела высоко достоверную положительную связь с содержанием CD4+FoxP3+ клеток. Корреляция выявлена между абсолютным количеством клеток, несущих маркер FoxP3, и РТК с фенотипом СD4+СD25+ high FoxP3+ (rpearson=0,86; p<0,001). В среднем РТК составляли 55% (95% ДИ 43–68) от абсолютного количества клеток, экспрессирующих FoxP3. Также отчетливая положительная взаимосвязь была обнаружена между общей экспрессией FoxP3+ в СD4+-лимфоцитах и РТК с фенотипом СD4+СD25+ highFoxP3+ (rpearson=0,95; p<0,001). Вместе с тем, не выявлено корреляции между содержанием СD4+СD25+ клеток и РТК (rpearson=–0,08; p=0,68). Соотношение РТК и всех клеток, экспрессирующих FoxP3 имело отчетливую тенденцию к росту по мере увеличения длительности периода после ТП (rpearson=0,42; p=0,024).У больных, обследованных в течение 1–3 мес. после трансплантации базиликсимаба, содержание CD25 на лимфоцитах и клеток с тройной меткой (СD4+СD25+ high FoxP3+) было крайне низким, приближаясь к нулю. В то же время, экспрессия CD4 и FoxP3 у этих больных сохранялась достаточно высокой, достоверно не отличаясь у больных, обследованных в более поздние сроки после операции. ЗАКЛЮЧЕНИЕ. Полученные данные косвенно свидетельствуют о том, что пул РТК после трансплантации почки, в значительной степени, определяется способностью лимфоцитов к экспрессии FoxP3. Причины обнаруженного снижения содержания РТК в циркуляции реципиентов ПТ, в частности, роль анти-CD25МАb, требуют проведения дальнейших исследований.

AIM OF THE STUDY. Regulatory T-cells (RTC) are one of mechanisms of immune tolerance development and support. The aim of this research was evaluation of the content of circulating RTC expressing markers CD4, CD25 and Foxp3 in patients with functioning kidney transplant (KT). PATIENTS AND METHODS. 30 stable KT recipients with duration of post-transplant care from 6 to 24 month (averagely 353 days (95%CI 246-459), 7 patients within 1-3 month after KT and 5 healthy persons were enrolled the observational study. All patients received anti-CD25 monoclonal antibodies induction (anti-CD25МАb – basiliximab). Research of CD4, CD25 and FoxP3 lymphocyte expression in peripheral blood was provided by flow cytrometry method. RESULTS. The proportion of CD4+CD25+high FoxP3+ cells from total CD4+ cells number varied widely and was significantly lower than in healthy persons. The total expression of FoxP3 in lymphocytes had highly significant positive correlation with CD4+FoxP3+ cells content. The correlation between absolute number of T-cells with marker FoxP3 and RTC with CD4+CD25 + high FOXP3+ phenotype was revealed (rpearson=0,86; p<0,001). Averagely percentage of RTC from absolute number of FoxP3 expressing cells was 55% (95%CI 43-68). Also positive correlation was revealed between common expression of FoxP3 in CD4+ lymphocytes and RTC with CD4+CD25 +high FOXP3+ phenotype (rpearson=0,95; p<0,001). No correlation was found between СD4+СD25+ cells and RTC(rpearson=-0,08; p=0,68). Ratio or RTC and all cells expressing FoxP3 had distinct tendency to growth with increase of post-transplant period (rpearson=0,42; p=0,024). In patients surveyed within 1-3 months after transplantation and basiliximab content of CD25 on lymphocytes and number of cells with triple mark (СD4+СD25+ high FoxP3+) was very low, approaching to zero. At the same time, expression of CD4 and FoxP3 was high enough in these patients, significantly without differing from patients surveyed in later terms after operation. CONCLUSION. Received data indirectly testify that RTC pool after kidney transplantation is mainly defined by lymphocytes ability to express FoxP3. Causes for decreased RTC content in KT recipients, particulary role of anti-CD25МАb, demand follow up studies.

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