bloodjourГематология и трансфузиологияRussian journal of hematology and transfusiology0234-57302411-3042ООО Издательский дом «Практика»10.35754/0234-5730-2020-65-4-417-430bloodjour-245Research ArticleОРИГИНАЛЬНЫЕ СТАТЬИORIGINAL ARTICLESОлигоклональность и субпопуляционный состав Т-клеток костного мозга у больных апластической анемиейOligoclonality and subpopulation structure of bone marrow T-cells in patients with aplastic anaemiahttps://orcid.org/0000-0002-8113-6115АбрамоваА. В.AbramovaA. V.

Абрамова Анастасия Владимировна, врач отделения высокодозной интенсивной химиотерапии гемобластозов и депрессий кроветворения с круглосуточным стационаром

125167, Москва

Anastasia V. Abramova, Physician, Department of High-Dose Intensive Chemotherapy of Hemoblastosis and Bone Marrow Depression

125167, Moscow

anastasia.abramova@list.ruhttps://orcid.org/0000-0002-8490-6066ГальцеваИ. В.GaltsevaI. V.

Гальцева Ирина Владимировна, кандидат медицинских наук, заведующая лабораторией иммунофенотипирования клеток крови и костного мозга

125167, Москва

Irina V. Galtseva, Cand. Sci. (Med.), Head of the Laboratory of Blood and Bone Marrow Cell Immunophenotyping

125167, Moscow

galtseva.i@blood.ruhttps://orcid.org/0000-0002-2449-2682МихайловаЕ. А.MikhailovaE. A.

Михайлова Елена Алексеевна, доктор медицинских наук, профессор, ведущий научный сотрудник отделения высокодозной интенсивной химиотерапии гемобластозов и депрессий кроветворения с круглосуточным стационаром

125167, Москва

Elena A. Mikhailova, Dr. Sci. (Med.), Professor, Leading Researcher, Department of High-Dose Intensive Chemotherapy of Hemoblastosis and Bone Marrow Depression

125167, Moscow

mikhailova.e@blood.ruhttps://orcid.org/0000-0002-6512-910XКапрановН. М.KapranovN. M.

Капранов Николай Михайлович, медицинский физик лаборатории иммунофенотипирования клеток крови и костного мозга

125167, Москва

Nikolay M. Kapranov, Medical Physicist, Laboratory of Blood and Bone Marrow Cell Immunophenotyping

125167, Moscow

kapranov.n@blood.ruhttps://orcid.org/0000-0001-5932-0285ДавыдоваЮ. О.DavydovaYu. O.

Давыдова Юлия Олеговна, врач клинической лабораторной диагностики лаборатории иммунофенотипирования клеток крови и костного мозга

125167, Москва

Yulia O. Davydova, Physician (clinical diagnostics), Laboratory of Blood and Bone Marrow Cell Immunophenotyping

125167, Moscow

juliya89mur@yandex.ruhttps://orcid.org/0000-0003-0934-6094ФидароваЗ. Т.FidarovaZ. T.

Фидарова Залина Таймуразовна, кандидат медицинских наук, заведующая отделением химиотерапии гемобластозов и депрессий кроветворения с дневным стационаром

125167, Москва

Zalina T. Fidarova, Cand. Sci. (Med.), Head of the Department of High-Dose Intensive Chemotherapy of Hemoblastosis and Bone Marrow Depression

125167, Moscow

zalinafidarova@mail.ruhttps://orcid.org/0000-0002-4827-8947ТроицкаяВ. В.TroitskayaV. V.

Троицкая Вера Витальевна, кандидат медицинских наук, заведующая отделением интенсивной высокодозной химиотерапии гемобластозов и депрессий кроветворения с круглосуточным стационаром; заместитель генерального директора по лечебной работе

125167, Москва

Vera V. Troitskaya, Cand. Sci. (Med.), Head of the Department of Intensive High-dose Chemotherapy of Hemoblastosis and Bone Marrow Depression with All-Day Hospital; Deputy Director for Medical Affairs

125167, Moscow

verat@blood.ruhttps://orcid.org/0000-0001-6177-3566ПаровичниковаЕ. Н.ParovichnikovaE. N.

Паровичникова Елена Николаевна, доктор медицинских наук, руководитель отдела химиотерапии гемобластозов, депрессий кроветворения и трансплантации костного мозга

125167, Москва

Elena N. Parovichnikova, Dr. Sci. (Med.), Head of the Department of Chemotherapy of Hemoblastosis, Hematopoiesis Depression and Bone Marrow Transplantation

125167, Moscow

elenap@blood.ruhttps://orcid.org/0000-0001-8188-5557СавченкоВ. Г.SavchenkoV. G.

Савченко Валерий Григорьевич, доктор медицинских наук, профессор, академик РАН, генеральный директор

125167, Москва

Valery G. Savchenko, Dr. Sci. (Med.), Full Member of the Russian Academy of Sciences, Professor, Director

125167, Moscow

svg@blood.ruФГБУ «Национальный медицинский исследовательский центр гематологии» Министерства здравоохранения Российской ФедерацииРоссияNational Research Center for HematologyRussian Federation202009122020654417430Copyright © Абрамова А.В., Гальцева И.В., Михайлова Е.А., Капранов Н.М., Давыдова Ю.О., Фидарова З.Т., Троицкая В.В., Паровичникова Е.Н., Савченко В.Г., 20202020Абрамова А.В., Гальцева И.В., Михайлова Е.А., Капранов Н.М., Давыдова Ю.О., Фидарова З.Т., Троицкая В.В., Паровичникова Е.Н., Савченко В.Г.Abramova A.V., Galtseva I.V., Mikhailova E.A., Kapranov N.M., Davydova Y.O., Fidarova Z.T., Troitskaya V.V., Parovichnikova E.N., Savchenko V.G.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.htjournal.ru/jour/article/view/245Ведение

Ведение. Основным патогенетическим механизмом развития апластической анемии (АА) считается нарушение иммунной регуляции кроветворения.

Цель

Цель: изучить субпопуляционный состав Т-клеток и репертуар Т-клеточного рецептора у больных АА.

Методы

Методы. C 2018 по 2020 г. в исследование были включены больные АА (n = 40) до начала иммуносупрессивной терапии. Исследование субпопуляционного состава Т-клеток и олигоклональности Т-клеточного рецептора по семействам Vβ (ТКР-Vβ) образцов костного мозга проводили с помощью метода проточной цитометрии.

Результаты

Результаты. Выявлены наиболее характерные особенности Т-клеточных субпопуляций у всех больных АА в образцах костного мозга: увеличение количества цитотоксических Т-клеток, эффекторных CD4+ и CD8+ клеток, CD4+ клеток памяти, что может подтверждать наличие длительной антигенной стимуляции с последующей активацией этих субпопуляций клеток, в результате которой происходит гиперэкспрессия провоспалительных цитокинов. Уменьшение наивных CD4+ и CD8+ клеток, регуляторных Т-клеток, двойных негативных Т-клеток может указывать на снижение контроля за цитокинпродуцирующими Т-клетками. Установлена связь между степенью тяжести АА и количеством эффекторных Т-клеток, Т-регуляторных клеток, двойных негативных Т-клеток и PD-1-позитивных Т-клеток. Самое большое количество потенциально цитокинпродуцирующих Т-клеток и минимальное количество клеток, участвующих в регуляции Т-клеточной активности, было выявлено у больных сверхтяжелой АА. При анализе репертуара ТКР-Vβ была обнаружена олигоклональная экспансия преимущественно в субпопуляции цитотоксических Т-клеток.

Заключение

Заключение. Обогащение определенных семейств Vβ свидетельствует о наличии аутореактивного Т-клеточного клона и подтверждает иммунный механизм развития АА. Динамическое исследование TКР-Vβ-репертуара может быть предложено в качестве мониторинга течения заболевания. Метод проточной цитофлуориметрии помогает выявить значимые биомаркеры для мониторинга клонов Т-клеток при АА с целью наиболее точной оценки активности патологического процесса при АА.

Introduction

Introduction. The main pathogenetic mechanism of the development of aplastic anemia (AA) is a violation of the immune regulation of hematopoiesis.

Aim

Aim: to study of the subpopulation composition of T-cells and the repertoire of the T-cell receptor in AA patients.

Patients and Methods

Patients and Methods. The study included AA patients (n = 40) without prior immunosuppressive therapy in 2018–2020. The T-cell subpopulation structure and T-cell receptor Vβ-family (TCR-Vβ) oligoclonality were studied in samples of bone marrow using flow cytometry.

Results

Results. We report characteristic properties of T-cell subpopulations of bone marrow in all AA patients: elevated counts of cytotoxic T-cells, effector CD4+ and CD8+ cells, CD4+ memory cells, which may suggest a long-term antigenic stimulation with subsequent activation of these cell subpopulations resulting in hyperexpression of pro-inflammatory cytokines. Diminishing of naive CD4+ and CD8+ cells, regulatory and double negative T-cells may indicate a relaxing control of cytokine-producing T-cells. A relationship has been established between the AA severity and counts of effector, regulatory, double negative and PD-1 positive T-cells. A highest count of potentially cytokine-producing T-cells and lowest count of cells involved in T-cell activity regulation were observed in very severe AA patients. Studies of the TCR-Vβ repertoire revealed oligoclonal expansion in the cytotoxic T-cell subpopulation.

Conclusion

Conclusion. Enrichment in selected Vβ families suggests autoreactive T-cell clonality and attests to the immune nature of AA. A dynamic TCR-Vβ repertoire assay may be recommended in the disease monitoring. Flow cytometry helps identify valuable biomarkers for T-cell clone monitoring in AA and a better assessment of the disease progression.

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