mimmunМедицинская иммунологияMedical Immunology (Russia)1563-06252313-741XSPb RAACI10.15789/1563-0625-PAF-2376mimmun-2376Research ArticleОРИГИНАЛЬНЫЕ СТАТЬИORIGINAL ARTICLESЭкспрессия поверхностных молекул и функциональные характеристики эндотелиальных клеток: влияние белковых фракций лизата микровезикул естественных киллеров в системе in vitroPhenotypic and functional characteristics of endothelial cells: the in vitro effects of protein fractions from the lysate of natural killer-derived microvesicleshttps://orcid.org/0000-0002-0365-8532КореневскийА. В.KorenevskyA. V.

Андрей Валентинович Кореневский – доктор биологических наук, ведущий научный сотрудник группы протеомной иммунорегуляции отдела иммунологии и межклеточных взаимодействий.

199034, Санкт-Петербург,  Менделеевская линия, 3. Тел.: 8 (812) 328-98-91, 323-75-45. Факс: 8 (812) 323-75-45

Andrey Valentinovich Korenevsky - PhD, MD (Biology), Leading Research Associate, Proteomic Immunoregulation Group, Department of Immunology and Cell Interactions.

199034, St. Petersburg,  Mendeleyevskaya Line, 3. Phone: 7 (812) 328-98-91, 323-75-45. Fax: 7 (812) 323-75-45

a.korenevsky@yandex.ruБерезкинаМ. Э.BerezkinaM. E.

Марьяна Эдуардовна Березкина – лаборант-исследователь лаборатории межклеточных взаимодействий отдела иммунологии и межклеточных взаимодействий.

Санкт-Петербург

Maryana Eduardovna Berezkina - Research Assistant, Laboratory of Cell Interactions, Department of Immunology and Cell Interactions.

St. Petersburg

mari-xari@list.ruГертТ. Н.GertT. N.

Татьяна Николаевна Герт – лаборант лаборатории межклеточных взаимодействий отдела иммунологии и межклеточных взаимодействий.

Санкт-Петербург

Tatyana Nikolayevna Gert - Assistant, Laboratory of Cell Interactions, Department of Immunology and Cell Interactions.

St. Petersburg

tniakimova94@gmail.comСинявинС. А.SinyavinS. A.

Серафим Александрович Синявин – студент группы протеомной иммунорегуляции отдела иммунологии и межклеточных взаимодействий.

Санкт-Петербург

Serafim Alexandrovich Sinyavin - Student, Proteomic Immunoregulation Group, Department of Immunology and Cell Interactions, D. Ott IOGRM.

St. Petersburg

sinyawin.serafim@mail.ruhttps://orcid.org/0000-0003-1560-7529СельковС. А.SelkovS. A.

Сергей Алексеевич Сельков – доктор медицинских наук, профессор, заслуженный деятель науки РФ, руководитель отдела иммунологии и межклеточных взаимодействий НИИАГР имени Д.О. Отта; профессор кафедры иммунологии Первый Санкт-Петербургский ГМУ имени академика И.П. Павлова.

Санкт-Петербург

Sergey Alexeyevich Selkov - PhD, MD (Medicine), Professor, Honored Researcher of Russia, Head, Department of Immunology and Cell Interactions, D. Ott IOGRM; Professor, Department of Immunology, First St. Petersburg State I. Pavlov MU.

St. Petersburg

selkovsa@mail.ruСоколовД. И.SokolovD. I.

Дмитрий Игоревич Соколов – доцент, заведующий лабораторией межклеточных взаимодействий отдела иммунологии и межклеточных взаимодействий НИИАГР имени Д.О. Отта; профессор кафедры иммунологии Первый Санкт-Петербургский ГМУ имени академика И.П. Павлова.

Санкт-Петербург

Dmitry Igorevich Sokolov - PhD, MD (Biology), Assistant Professor, Head, Laboratory of Cell Interactions, Department of Immunology and Cell Interactions, D. Ott IOGRM; Professor, Department of Immunology, First St. Petersburg State I. Pavlov MU.

St. Petersburg

falcojugger@yandex.ruФГБНУ Научно-исследовательский институт акушерства, гинекологии и репродуктологии имени Д.О. ОттаРоссияD. Ott Institute of Obstetrics, Gynecology, and Reproductive MedicineRussian FederationФГБНУ Научно-исследовательский институт акушерства, гинекологии и репродуктологии имени Д.О. Отта; ФГБОУ ВО Первый Санкт-Петербургский государственный медицинский университет имени академика И.П. Павлова Министерства здравоохранения РФРоссияD. Ott Institute of Obstetrics, Gynecology, and Reproductive Medicine; First St. Petersburg State I. Pavlov Medical UniversityRussian Federation202213072022243463480Copyright © Кореневский А.В., Березкина М.Э., Герт Т.Н., Синявин С.А., Сельков С.А., Соколов Д.И., 20222022Кореневский А.В., Березкина М.Э., Герт Т.Н., Синявин С.А., Сельков С.А., Соколов Д.И.Korenevsky A.V., Berezkina M.E., Gert T.N., Sinyavin S.A., Selkov S.A., Sokolov D.I.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/2376

Микровезикулы – мембранные образования размером от 100 до 1000 нм, продуцируемые различными клетками в состоянии покоя и активации, – могут передавать компоненты своего содержимого клеткам-мишеням, регулировать физиологические процессы, участвовать в развитии патологий. Среди множества клеток-источников микровезикул особый интерес представляют естественные киллеры – субпопуляция лимфоцитов, осуществляющих контактный цитолиз вирус-инфицированных и опухолевых клеток, а также участвующих в регуляции ангиогенеза. Продуцируя различные стимуляторы и ингибиторы этого процесса, естественные киллеры способны изменять функциональную активность эндотелиальных клеток путем контактного взаимодействия с ними собственных микровезикул. Учитывая недостаточность имеющихся в литературе сведений о способности экстраклеточных везикул влиять на функциональное состояние эндотелия в зависимости от баланса передаваемых ими про- и антиангиогенных факторов, целью данного исследования явилось изучение влияния белковых фракций лизата микровезикул, продуцируемых клетками естественных киллеров линии NK 92, на фенотип и функциональные характеристики эндотелиальных клеток линии EA.hy926 в модельном эксперименте in vitro. В результате микропрепаративного разделения лизата микровезикул клеток линии NK-92 было получено двенадцать белковых фракций (индукторы). Установлено, что пролиферация и миграция клеток линии EA.hy926 после их культивирования в присутствии десяти из двенадцати полученных фракций, в зависимости от концентрации содержащихся в них активных компонентов, изменялись разнонаправленно и дозозависимо или оставались неизменными. Показан преимущественно стимулирующий эффект воздействия индукторов на пролиферацию клеток-мишеней, что свидетельствует о наличии в этих фракциях белков, регулирующих функции эндотелия. При этом остаточная площадь, не занятая мигрировавшими эндотелиальными клетками после их культивирования в присутствии индукторов, не всегда коррелировала с интенсивностью миграции и не была обратно пропорциональна количеству мигрировавших клеток. Дополнительно было установлено, что ни одна из полученных белковых фракций не оказывала влияния на экспрессию рецепторов CD54 (ICAM-1), CD34, CD31 (PECAM-1), CD119 (IFNγR1) клетками линии EA.hy926. Полученные данные об изменении функциональных характеристик клеток линии EA.hy926 под влиянием белковых фракций лизата микровезикул, продуцируемых клетками линии NK-92, подтверждают вовлеченность этих субклеточных образований в обеспечение коммуникации естественных киллеров с клетками эндотелия и указывают на различное участие эффекторных белков, переносимых микровезикулами, в механизмах ангиогенеза.

Microvesicles are membrane-derived formations ranging in size from 100 to 1000 nm, being produced by a variety of resting and activated cells. They can transfer their cargo to target cells, regulate physiological processes, and participate in the development of clinical disorders. Among the microvesicles of different origin, natural killers are of special interest. They represent a subpopulation of lymphocytes that eliminate aberrant cells, including virally infected and malignant cells, and participate in regulation of angiogenesis. By producing various stimuli and inhibitors of the latter process, natural killers are able to change functional activity of endothelial cells by means of microvesicle-mediated contacts. There are only scarce literature data on ability of the extracellular vesicles to influence endothelial functions, depending on the intrinsic balance of pro- and anti-angiogenic factors. Therefore, the aim of our study was to evaluate the effect of protein fractions derived from microvesicle lysate of the NK-92 natural killer cell line upon phenotype and functional characteristics of EA.hy926 endothelial cell line under in vitro experimental conditions. Using chromatographic micro-preparatory separation, twelve protein fractions (inducers) were obtained from the lysate. It was found that proliferation and migration of EA.hy926 cells after their cultivation with 10 of 12 protein fractions, were changed in different directions. These effects were dose-dependent, or remained unchanged, at distinct concentrations of active components in the fractions. The inducing factors from these fractions exerted predominantly stimulating effects on proliferation of the target cells, thus suggesting presence of proteins which are able of regulating endothelial functions. However, the size of residual area free of migrating endothelial cells treated by the inducers did not always correlate with the migration intensity and did not inversely correlate with the number of migrating cells. Moreover, it was found that the obtained protein fractions had no effect upon expression of CD54 (ICAM-1), CD34, CD31 (PECAM-1) and CD119 (IFNγR1) receptors by EA.hy926 cells. The data obtained confirm an involvement of microvesicles in communications between natural killer cells and endothelial cells, and presume different participation modes of microvesicle-derived effector proteins in the angiogenesis machinery.

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