medsovetМедицинский СоветMeditsinskiy sovet = Medical Council2079-701X2658-5790REMEDIUM GROUP Ltd.10.21518/ms2025-271medsovet-9376Research ArticleПРАКТИКАPRACTICEКомплексный анализ субпопуляций В-клеток и фолликулярных Т-клеток в небных миндалинах человекаComprehensive analysis of B cell and follicular T cell subpopulations in human palatine tonsilshttps://orcid.org/0009-0003-7962-2389КухарчукА. А.KuharchukA. A.

Кухарчук Артем Алексеевич, лаборант-исследователь отдела молекулярных технологий Научно-исследовательского института трансляционной медицины 

117997, Москва, ул. Островитянова, д. 1

Artem А. Kuharchuk, Laboratory Assistant Researcher at the Department of Molecular Technologies at the Translational Medicine Research Institute

1, Ostrovityanov St., Moscow, 117997

kuharchukartem@yandex.ruhttps://orcid.org/0000-0003-1699-9237СоколоваС. Р.SokolovaS. R.

Соколова София Романовна, младший научный сотрудник отдела молекулярных технологий Научно-исследовательского института трансляционной медицины 

117997, Москва, ул. Островитянова, д. 1

Sofia R. Sokolova, Researcher of Molecular Technologies Department of Institute of Translational Medicine 

1, Ostrovityanov St., Moscow, 117997

 

sophiya.sokolova@gmail.comhttps://orcid.org/0009-0004-4397-9404ХалифаеваД. З.KhalifaevaJ. Z.

Халифаева Джамиля Зиядиновна, аспирант кафедры оториноларингологии имени академика Б.С. Преображенского Института хирургии, Российский национальный исследовательский медицинский университет имени Н.И. Пирогова; врач-оториноларинголог оториноларингологического отделения, Городская клиническая больница №1 имени Н.И. Пирогова

117997, Москва, ул. Островитянова, д. 1; 117049, Москва, Ленинский проспект д. 8

Jamilya Z. Khalifaeva, Postgraduate Student of the Department of Otorhinolaryngology named after Academician B.S. Preobrazhensky Institute of Surgery; Otorhinolaryngologist of the Otorhinolaryngological Department 

1, Ostrovityanov St., Moscow, 117997; 18a, Bldg. 2, Zagorodnoe Shosse, Moscow, 117152

 

Khalifaieva96@mail.ruhttps://orcid.org/0000-0001-9811-8397ГуровА. В.GurovA. V.

Гуров Александр Владимирович, д.м.н., профессор кафедры оториноларингологии имени академика Б.С. Преображенского Института хирургии, профессор кафедры микробиологии и вирусологии Института профилактической медицины, Российский национальный исследовательский медицинский университет имени Н.И. Пирогова;  старший научный сотрудник, Научно-исследовательский клинический институт оториноларингологии имени Л.И. Свержевского

117997, Москва, ул. Островитянова, д. 1; 117152, Москва, Загородное шоссе, д. 18а, стр. 2

Alexander V. Gurov, Dr. Sci. (Med.), Professor of the Department of Otorhinolaryngology named after Academician B.S. Preobrazhensky Institute of Surgery, Professor of the Department of Microbiology and Virology of the Institute of Preventive Medicine, Pirogov Russian National Research Medical University; Senior Researcher, Sverzhevsky Scientific Research Clinical Institute of Otorhinolaryngology

1, Ostrovityanov St., Moscow, 117997; 18a, Bldg. 2, Zagorodnoe Shosse, Moscow, 117152

Alex9999@inbox.ruhttps://orcid.org/0000-0002-4963-7403ГригороваИ. Л.GrigorovaI. L.

Григорова Ирина Леонидовна, к.б.н., старший научный сотрудник отдела молекулярных технологий Научно-исследовательского института трансляционной медицины 

117997, Москва, ул. Островитянова, д. 1

Irina L. Grigorova, Cand. Sci. (Biol.), Senior Researcher of Molecular Technologies Department of Institute of Translational Medicine 

1, Ostrovityanov St., Moscow, 117997

grig76@gmail.comРоссийский национальный исследовательский медицинский университет имени Н.И. ПироговаРоссияPirogov Russian National Research Medical UniversityRussian FederationРоссийский национальный исследовательский медицинский университет имени Н.И. Пирогова; Городская клиническая больница №1 имени Н.И. ПироговаРоссияPirogov Russian National Research Medical University; City Clinical Hospital No. 1 named after N.I. PirogovRussian FederationРоссийский национальный исследовательский медицинский университет имени Н.И. Пирогова; Научно-исследовательский клинический институт оториноларингологии имени Л.И. СвержевскогоРоссияPirogov Russian National Research Medical University; Sverzhevsky Scientific Research Clinical Institute of OtorhinolaryngologyRussian Federation2025100920251913289297Copyright © Кухарчук А.А., Соколова С.Р., Халифаева Д.З., Гуров А.В., Григорова И.Л., 20252025Кухарчук А.А., Соколова С.Р., Халифаева Д.З., Гуров А.В., Григорова И.Л.Kuharchuk A.A., Sokolova S.R., Khalifaeva J.Z., Gurov A.V., Grigorova I.L.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.med-sovet.pro/jour/article/view/9376Введение

Введение. Небные миндалины играют ключевую роль в защите организмa человека от многочисленных патогенов, попадающих через верхние дыхательные пути и пищеварительный тракт. Одной из основных функций небных миндалин является формирование гуморального иммунитета, который возникает в результате В-клеточного ответа на чужеродные антигены при участии и под контролем фолликулярных Т-клеток. Нарушения в вовлечении Ви Т-клеток в иммунный ответ в миндалинах могут быть связаны с различными патологиями, хроническими инфекциями, иммунодефицитами или лимфопролиферативными заболеваниями.

Цель

Цель. Провести методический анализ различных субпопуляций В-клеток и Т-фолликулярных клеток в миндалинах с помощью проточной цитометрии. Оценить различные компоненты локального иммунного ответа.

Материалы и методы

Материалы и методы. Отбор пациентов для исследования проводили в оториноларингологическом отделении городской клинической больницы №1 им. Н.И. Пирогова г. Москвы с 2024 по 2025 г. В качестве материала использовали миндалины, полученные в ходе тонзиллэктомии. Выделенные из органов мононуклеарные клетки (МНК) окрашивали смесью поверхностных маркеров В-клеток и Т-фолликулярных клеток, а также проводили внутриклеточное окрашивание транскрипционных факторов Bcl6 и FoxP3 для дальнейшего анализа методом проточной цитометрии.

Результаты и обсуждение

Результаты и обсуждение. На основе полученных данных была предложена стратегия выделения основных субпопуляций В-клеток (наивных, двойных негативных В-клеток, клеток памяти, В-клеток герминативных центров (ГЦ)), реализующих гуморальный иммунный ответ в миндалинах. Кроме этого, был разработан подход для анализа Т-фолликулярных клеток в миндалинах с выделением субпопуляций Т-фолликулярных хелперных (Тфх), регуляторных (Тфр) клеток, а также CD25++ Тфх, которые необходимые для контроля и регуляции В-клеточных ответов.

Выводы

Выводы. Разработанный анализ может быть использован как в фундаментальных исследованиях, так и в клинической практике для диагностики и мониторинга иммунопатологических состояний.

Introduction

Introduction. Palatine tonsils play an important role in protecting the human body from numerous pathogens entering through the upper respiratory and digestive tracts. One of the main components of the protective function of the palatine tonsils is the formation of humoral immunity, which occurs as a result of a B-cell response to foreign antigens with the participation and under the control of follicular T cells. Disorders in the involvement of B and T cells in the immune response in the tonsils may be associated with various pathologies, chronic infections, immunodeficiency or lymphoproliferative diseases.

Aim

Aim. To perform a methodical analysis of various subpopulations of B and follicular T cells in tonsils using flow cytometry. To evaluate various components of the local immune response.

Materials and methods

Materials and methods. The selection of patients for the study was carried out in the otorhinolaryngological department of the City Clinical Hospital No. 1 named after N. I. Pirogov, Moscow from 2024 to 2025. Fresh tonsils were obtained from patients undergoing tonsillectomy or tonsillotomy. Mononuclear cells (MNCs) isolated from the organs were stained with a mixture of surface markers for Band T-follicular cells, followed by intracellular staining of the transcription factors Bcl6 and FoxP3 for subsequent flow cytometry analysis.

Results and discussion

Results and discussion. Based on the obtained data, a strategy was proposed for identifying key B-cell subpopulations (naïve, double-negative B cells, memory B cells, and germinal center (GC) B cells) involved in the humoral immune response in tonsils. Additionally, an approach was developed for analyzing T-follicular cells in tonsils, distinguishing subpopulations of T-follicular helper (Tfh) cells, T-follicular regulatory (Tfr) cells, and CD25++ Tfh cells, which are essential for the control and regulation of B-cell responses.

Conclusions

Conclusions. The developed analysis can be used both in basic research and in clinical practice for the diagnosis and monitoring of immunopathological conditions.

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