mimmunМедицинская иммунологияMedical Immunology (Russia)1563-06252313-741XSPb RAACI10.15789/1563-0625-2019-3-503-516mimmun-1824Research ArticleОРИГИНАЛЬНЫЕ СТАТЬИORIGINAL ARTICLESСОДЕРЖАНИЕ CD4+ И CD8+ ЭФФЕКТОРНЫХ КЛЕТОК ПАМЯТИ И ПРОЛИФЕРАТИВНАЯ АКТИВНОСТЬ Т-ЛИМФОЦИТОВ ПРИ БРОНХИАЛЬНОЙ АСТМЕCONTENTS OF CD4+ AND CD8+ EFFECTOR MEMORY CELLS AND PROLIFERATIVE ACTIVITY OF T LYMPHOCYTES IN BRONCHIAL ASTHMAБарковскаяМ. Ш.BarkovskayaM. Sh.

младший научный сотрудник лаборатории клинической иммунопатологии

630099, г. Новосибирск, ул. Ядринцевская, 14.Тел.: 8 (383) 227-01-35. Тел./факс: 8 (383) 222-70-28.

MD, Junior Research Associate, Laboratory of Clinical Immunopathology

630099, Novosibirsk, Yadrintsevskaya str., 14.Phone: 7 (383) 227-01-35. Phone/Fax: 7 (383) 222-70-28.

margaritabarkovskaya@gmail.comБлиноваЕ. А.BlinovaE. A.

к.б.н., старший научный сотрудник лаборатории клинической иммунопатологии

г. Новосибирск

PhD (Biology), Senior Research Associate, Laboratory of Clinical Immunopathology

Novosibirsk

ГришинаЛ. В.GrishinaL. V.

к.б.н., научный сотрудник лаборатории клинической иммунопатологии

г. Новосибирск

PhD (Biology), Research Associate, Laboratory of Clinical Immunopathology

Novosibirsk

ЛеоноваМ. И.LeonovaM. I.

врач аллерголог-иммунолог отделения аллергологии

г. Новосибирск

MD, Allergologist-Immunologist, Department of Allergology

Novosibirsk

НепомнящихВ. М.NepomniashchikchV. M.

врач аллерголог-иммунолог отделения аллергологии

г. Новосибирск

MD, Allergologist-Immunologist, Department of Allergology

Novosibirsk

ДеминаД. В.DeminaD. V.

к.м.н., врач аллерголог-иммунолог, заведующий отделением аллергологии

г. Новосибирск

MD, PhD (Medicine), Allergologist-Immunologist, Head, Department of Allergology

Novosibirsk

КозловВ. А.KozlovV. A.

д.м.н., профессор, академик РАН, заведующий лабораторией клинической иммунопатологии, научный руководитель

г. Новосибирск

MD, DC (Medicine), Professor, Full Member, Russian Academy of Sciences, Head, Laboratory of Clinical Immunopathology, Scientific Director

Novosibirsk

ФГБНУ «Научно-исследовательский институт фундаментальной и клинической иммунологии»РоссияResearch Institute of Fundamental and Clinical ImmunologyRussian Federation201912072019213503516Copyright © Барковская М.Ш., Блинова Е.А., Гришина Л.В., Леонова М.И., Непомнящих В.М., Демина Д.В., Козлов В.А., 20192019Барковская М.Ш., Блинова Е.А., Гришина Л.В., Леонова М.И., Непомнящих В.М., Демина Д.В., Козлов В.А.Barkovskaya M.S., Blinova E.A., Grishina L.V., Leonova M.I., Nepomniashchikch V.M., Demina D.V., Kozlov 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://www.mimmun.ru/mimmun/article/view/1824

Бронхиальная астма – хроническое воспалительное заболевание дыхательных путей, в патогенезе которого ключевая роль отводится Т-лимфоцитам. Считается, что сокращение числа наивных Т-клеток и накопление Т-лимфоцитов памяти при бронхиальной астме сопровождаются функциональными изменениями Т-лимфоцитов. В данном исследовании был изучен субпопуляционный состав Т-лимфоцитов в периферической крови, в нестимулированной и PHA-стимулированной культурах, а также пролиферативная активность Т-клеточных субпопуляций у пациентов с бронхиальной астмой и здоровых доноров. В исследование было включено 10 пациентов с бронхиальной астмой (возраст 45,4±11,8 лет). Одна половина пациентов находилась в стадии ремиссии, вторая половина – в стадии обострения основного заболевания. Доноры подбирались по полу и возрасту. Методом проточной цитофлуориметрии по экспрессии маркеров CD45R0, CD62L и CD197(CCR7)CD4+ и CD8+Т-лимфоциты были разделены на центральные (Tcm) и эффекторные клетки памяти (Tem), наивные Т-лимфоциты (Tnaïve) и терминально-дифференцированные эффекторы (Temra). В качестве функционального показателя была исследована пролиферативная активность Тcm, Tem и Tnaïve в ответ на PHA. В ходе исследования было установлено, что содержание CD4+ТemCD62L+ и CD8+ТemCD62L+ в периферической крови у пациентов в стадии обострения астмы было значимо снижено по сравнению с донорами. При стимуляции PHA данные различия в субпопуляционном составе между группами пациентов и доноров нивелировались. Также было обнаружено наличие положительной корреляции относительного количества CD4+ и CD8+ клеток памяти с возрастом у пациентов с бронхиальной астмой, которая отсутствовала у доноров. Анализ митогениндуцированной пролиферации показал, что наиболее активно делились Tcm и Tnaïve клетки как у пациентов, так и у доноров. При этом пролиферативная активность CD4+Т-лимфоцитов и субпопуляций CD8+Tcm, CD4+Tcm и CD4+Tem62L- была выше в группе пациентов с ремиссией астмы, чем в группах пациентов, находящихся в обострении, и здоровых доноров. Выявленное увеличение относительного количества Т-клеток памяти с возрастом позволяет предполагать их участие в процессе развития бронхиальной астмы. Сопоставимая с донорскими значениями пролиферативная реакция изученных субпопуляций говорит о сохранности функциональных свойств Т-клеток памяти и наивных Т-лимфоцитов при бронхиальной астме. Усиление пролиферации отдельных супопуляций у пациентов, находящихся в ремиссии бронхиальной астмы свидетельствует об активированном статусе Т-клеток памяти. Обнаруженное снижение количества CD4+ТemCD62L+ и CD8+ТemCD62L+ у пациентов в обострении, на наш взгляд, может быть связано с активным воспалительным процессом в бронхиальном дереве.

Bronchial asthma is a chronic inflammatory disease of the respiratory tract. T-lymphocytes play a key role in pathogenesis of this allergic disease. The reduction in number of naïve T cells and the accumulation of memory T cells in bronchial asthma are accompanied by dysregulation of T lymphocyte function. In present study, we have investigated the contents of different T lymphocyte subpopulations in peripheral blood as well as in resting and PHA-stimulated cultures, along with their proliferative capacity in patients with bronchial asthma and healthy donors. The study included 10 patients with bronchial asthma (age 45.4±11.8 years). One-half of patients was in remission state, the others having been at the stage of clinical exacerbation. The group of donors was formed by healthy individuals matched by gender and age to the patients. Based on expression of cell surface markers CD45R0, CD62L and CD197 (CCR7), the CD4+ and CD8+T lymphocytes were divided into central (Tcm) and effector memory cells (Tem), naïve T lymphocytes (Tnaïve) and terminally differentiated effector cells (Temra) using flow cytometry technique. The proliferative activity of Tcm, Tem and Tnaïve was evaluated in response to PHA as a functional marker of T cells. We have found that the percentage of peripheral CD4+TemCD62L+ and CD8+TemCD62L+ cells in the patients with asthma exacerbation was significantly reduced, if compared to the donors. Following PHA stimulation, these differences in T cell subsets between the groups of patients and donors were not detectable. We performed a correlation analysis between the memory T cell contents and age of the subjects studied. It was shown that the relative amounts of CD4+ and CD8+ memory cells increased with age in asthmatics, but not in healthy donors. Analysis of mitogen-induced proliferation showed that Tcm and Tnaïve cells proliferated more actively than other subpopulations in both groups. Meanwhile, the proliferative activity of CD4+T lymphocytes and subsets of CD8+Tcm, CD4+Tcm and CD4+Tem62L was higher in the group of asthma patients in remission state than in the patients with exacerbating disease, and healthy donors. The revealed increase in the relative number of memory T cells with age suggests that these cells participate in development of bronchial asthma. Proliferative response of the studied subpopulations, which was comparable to the donor values, suggests a functional maintenance of memory T cells and naïve T lymphocytes in bronchial asthma. The increased proliferation of some T-cell subpopulations in asthmatics in remission suggests an activated state of memory T cells. The observed decrease in the number of CD4+TemCD62L+ and CD8+TemCD62L+ in patients with asthma exacerbation may be, by our opinion, associated with an active inflammatory process in the airways.

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