mimmunМедицинская иммунологияMedical Immunology (Russia)1563-06252313-741XSPb RAACI10.15789/1563-0625-MPO-2920mimmun-2920Research ArticleОРИГИНАЛЬНЫЕ СТАТЬИORIGINAL ARTICLESМинорная популяция NK-лимфоцитов с коэкспрессией CD19Minor population of NK lymphocytes with CD19 coexpressionКалашниковаА. А.KalashnikovaA. A.

Калашникова А.А. – к.б.н., старший научный сотрудник научно-исследовательского отдела лабораторной диагностики научно-исследовательского центра

197345, Россия, Санкт-Петербург, ул. Лебедева, 4/2

Тел.: 8 (812) 339-39-39

Kalashnikova A.A., PhD (Biology), Senior Research Associate, Research Department of Laboratory Diagnostics

4/2 Lebedev St St. Petersburg 197345 Russian Federation

Phone: +7 (812) 339-39-39

petkova_nas@mail.ruБычковаН. В.BychkovaN. V.

Бычкова Н.В. – д.б.н., ведущий научный сотрудник научно-исследовательского отдела лабораторной диагностики научно-исследовательского центра; доцент кафедры иммунологии

Санкт-Петербург

Bychkova N.V., PhD, MD (Biology), Leading Research Associate, Research Department of Laboratory Diagnostics;  Associate Professor, Department of Immunology 

St. Petersburg

ФГБУ «Всероссийский центр экстренной и радиационной медицины имени А.М. Никифорова» МЧС РоссииРоссияA. Nikiforov Russian Centre of Emergency and Radiation MedicineRussian FederationФГБУ «Всероссийский центр экстренной и радиационной медицины имени А.М. Никифорова» МЧС России; ФГБОУ ВО «Санкт-Петербургский государственный медицинский университет имени академика И.П. Павлова» Министерства здравоохранения РФРоссияA. Nikiforov Russian Centre of Emergency and Radiation Medicine; St. Petersburg State I. Pavlov Medical UniversityRussian Federation202407032024263513522Copyright © Калашникова А.А., Бычкова Н.В., 20242024Калашникова А.А., Бычкова Н.В.Kalashnikova A.A., Bychkova N.V.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/2920

В последнее десятилетие в связи с широким распространением многоцветной проточной цитометрии для рутинных тестов в зарубежной литературе появились единичные сообщения о выявлении в крови и костном мозге пациентов минорной субпопуляции NK-клеток со слабой коэкспрессией В-клеточного антигена CD19. Практически отсутствует оценка частоты встречаемости и относительного количества CD56+CD19+dim клеток, нет данных о фенотипических особенностях, а также связи этой субпопуляции с какой-либо патологией. Цель исследования – оценить частоту встречаемости, относительное количество и фенотипические характеристики минорной субпопуляции лимфоцитов CD56+CD19+dim в образцах крови пациентов, направляемых на исследование субпопуляционного состава лимфоцитов. Материалом являлась периферическая кровь иммунокомпрометированных лиц. Методом восьмицветной проточной цитометрии определяли субпопуляционный состав лимфоцитов с использованием маркеров CD3, CD4, CD8, CD19, CD25, CD45, CD56, HLA-DR. Для оценки частоты встречаемости субпопуляции CD56+CD19+dim осуществляли ретроспективный анализ LMD-файлов 1210 исследований для 935 пациентов. Средний возраст обследованных лиц 39,8±14,7 года, среди них 84 ребенка до 18 лет. Ряду пациентов исследование выполнялось неоднократно. Дополнительное фенотипирование CD56+CD19+dim клеток проводили с использованием широкой панели антител к В-клеточным и T/NK-клеточным антигенам. Частота встречаемости образцов крови, содержащих CD56+CD19+dim клетки, составила 1,2%, при относительном количестве субпопуляции 2,1±1,9% от лимфоцитов и 0,8±0,6% от лейкоцитов. Максимальный размер субпопуляции составил 8,8% от лимфоцитов (2,8% от лейкоцитов). Отмечено длительное сохранение субпопуляции на протяжении всего периода наблюдения за пациентами – от двух месяцев до 6 лет. Сопоставление экспрессии дополнительных маркеров субпопуляциями NK-лимфоцитов CD56+CD19+dim и CD56+CD19- выявило особенности первой, а именно высокую экспрессию CD2 и CD57, сниженную плотность экспрессии CD7, CD16, CD38. Определен фенотип изученной субпопуляции: CD56+dimCD19+dimCD2+brightCD7+dimCD11c+CD16+dimCD38+dimCD45RA+CD57+CD94+dimNKG2D+CD3-CD4-CD5-CD20-CD21-CD25-CD45R0-CD62L-CD79b-CD117- с вариабельной экспрессией CD8 и HLA-DR. Фенотип соответствует активированным терминально дифференцированным адаптивным NK-лимфоцитам, связанным с цитомегаловирусной инфекцией. В нашей работе у лиц с субпопуляцией CD56+CD19+dim лимфоцитов в крови отмечена цитомегаловирусная инфекция в анамнезе и реактивация хронической ВЭБ-инфекции на момент исследования. Вероятной причиной коэкспрессии CD19 может быть трогоцитоз NK-клеткой фрагмента мембраны В-лимфоцита при активной ВЭБ-инфекции. Cубпопуляция CD56+CD19+dim лимфоцитов может достигать заметных величин и искажать результаты иммунологических исследований, выполняемых методом проточной цитометрии. Особенно вероятны ошибки при оценке минимальной определяемой болезни при острых В-клеточных лейкозах. Минорная популяция NK-лимфоцитов CD56+CD19+dim может быть идентифицирована в рутинных иммунологических исследованиях, а ее функциональные особенности и связь с патологией нуждаются в дальнейшем изучении.

Single reports were published concerning a minor subpopulation of NK cells with weak coexpression of the B cell antigen CD19 in the patients’ blood and bone marrow. The frequency and relative number of CD56+CD19+dim cells is virtually not assessed, and there is no data on their phenotypic characteristics, as well as the connection of this subpopulation with any disease state. The purpose of the present study was to assess the frequency, relative quantity and phenotypic characteristics of CD56+CD19+dim lymphocytes in blood of patients referred for assessment of the lymphocyte subpopulation profile. Peripheral blood of immunocompromised individuals was taken, and subpopulation composition of lymphocytes was determined using eight-color flow cytometry (markers: CD3, CD4, CD8, CD19, CD25, CD45, CD56, HLA-DR). To estimate incidence of the CD56+CD19+dim subpopulation, we have carried out a retrospective analysis of LMD files on 1210 studies for 935 patients (average age, 39.8±14.7 years old) including 84 children under 18 years old. The study was performed repeatedly for some patients. Phenotyping of CD56+CD19+dim cells was performed using a panel of antibodies to B cell, T/NK cell antigens. The occurrence of blood samples containing CD56+CD19+dim was 1.2%, with a relative content of 2.1±1.9% among total lymphocyte population (0.8±0.6% of leukocytes). Long-term persistence of the subpopulation was noted in the patients throughout the entire observation period. The comparison of specific marker expression by NK CD56+CD19+dim, and CD56+CD19- cells revealed high expression of CD2, CD57, reduced expression density of CD7, CD16, CD38. The phenotype of the studied NK cell subpopulation was as follows: CD56+dimCD19+dimCD2+brightCD7+dimCD11c+CD16+dimCD38+dimCD45RA+CD57+CD94+dimNKG2D+CD3-CD4-CD5-CD20-CD21-CD25-CD45R0-CD62L-CD79b-CD117-, with variable expression of CD8 and HLA-DR. The phenotype is consistent with activated terminally differentiated adaptive NK associated with cytomegalovirus infection. The individuals with CD56+CD19+dim had a history of CMV-infection and reactivation of chronic EBV-infection at the time of the study. A probable cause of CD19 coexpression may be trogocytosis of B cell membrane fragments by natural killer cells during active EBV-infection. CD56+CD19+dim lymphocytes can reach noticeable values thus altering the results of studies performed by flow cytometry. The errors are most likely to occur upon assessing the minimal residual disease levels in acute B cell leukemias. The minor CD56+CD19+dimNK subpopulation may be detected in routine immunological analysis. Its functional features and association with certain disorders require further studies.

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