Abstract
Introduction. The main pathogenetic mechanism of the development of aplastic anemia (AA) is a violation of the immune regulation of hematopoiesis.Aim: to study of the subpopulation composition of T-cells and the repertoire of the T-cell receptor in AA patients.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. 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. 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.
Highlights
The main pathogenetic mechanism of the development of aplastic anemia (AA) is a violation of the immune regulation of hematopoiesis
Границы референсных интервалов для каждого из 24 клонов были рассчитаны на основании анализа костном мозге (КМ) доноров и включали значения от 2,5 до 97,5 процентиля после исключения выбросов
Main T-cell subpopulations and phenotypes defined in the study
Summary
В качестве контрольной группы для определения субпопуляций T-клеток и экспансии клонов Vβ Т-клеток использовали КМ 23 здоровых доноров КМ, подписавших информированное согласие на включение в исследование. Для анализа ТКР-Vβ-репертуара Т-лимфоцитов КМ больных (n = 39) (данные одного пациента были удалены из исследования вследствие ошибки подготовки пробы) был использован коммерческий набор IOTest Beta Mark ТКР-Vβ Repertoire (Beckman Coulter, Майами, Флорида, США), позволяющий оценить следующие семейства ТКР-Vβ: Vβ 1, Vβ 2, Vβ 3, Vβ 4, Vβ 5.1, Vβ 5.2, Vβ 5.3 , Vβ 7.1, Vβ 7.2, Vβ 8, Vβ 9, Vβ 11, Vβ 12, Vβ 13.1, Vβ 13.2, Vβ 13.6, Vβ 14, Vβ 16, Vβ 17, Vβ 18, Vβ 20, Vβ 21.3, Vβ 22 и Vβ 23. Границы референсных интервалов для каждого из 24 клонов были рассчитаны на основании анализа КМ доноров и включали значения от 2,5 до 97,5 процентиля после исключения выбросов. Для определения отличий в долях различных субпопуляций T-клеток у больных и доноров использовали. Monoclonal antibodies to human differentiation antigens used in the study
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