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<article article-type="research-article" dtd-version="1.3" xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" xml:lang="ru"><front><journal-meta><journal-id journal-id-type="publisher-id">mimmun</journal-id><journal-title-group><journal-title xml:lang="ru">Медицинская иммунология</journal-title><trans-title-group xml:lang="en"><trans-title>Medical Immunology (Russia)</trans-title></trans-title-group></journal-title-group><issn pub-type="ppub">1563-0625</issn><issn pub-type="epub">2313-741X</issn><publisher><publisher-name>SPb RAACI</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="doi">10.15789/1563-0625-UCB-2846</article-id><article-id custom-type="elpub" pub-id-type="custom">mimmun-2846</article-id><article-categories><subj-group subj-group-type="heading"><subject>Research Article</subject></subj-group><subj-group subj-group-type="section-heading" xml:lang="ru"><subject>КРАТКИЕ СООБЩЕНИЯ</subject></subj-group><subj-group subj-group-type="section-heading" xml:lang="en"><subject>SHORT COMMUNICATIONS</subject></subj-group></article-categories><title-group><article-title>Пуповинная кровь как перспективный источник NK-клеток для иммунотерапии</article-title><trans-title-group xml:lang="en"><trans-title>Umbilical cord blood as a promising source of NK cells for immunotherapy</trans-title></trans-title-group></title-group><contrib-group><contrib contrib-type="author" corresp="yes"><contrib-id contrib-id-type="orcid">https://orcid.org/0000-0002-7209-1373</contrib-id><name-alternatives><name name-style="eastern" xml:lang="ru"><surname>Величинский</surname><given-names>Р. А.</given-names></name><name name-style="western" xml:lang="en"><surname>Velichinskii</surname><given-names>R. A.</given-names></name></name-alternatives><bio xml:lang="ru"><p>Величинский Родион Альбертович — аспирант, инженер-исследователь.</p><p>117997, Москва, ул. Миклухо-Маклая, 16/10</p><p>Тел.: 8 (977) 849-29-34</p></bio><bio xml:lang="en"><p>Rodion A. Velichinskii - Postgraduate Student, Engineer-Researcher, Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences.</p><p>16/10 Miklukho-Maklay St Moscow 117997</p><p>Phone: +7 (977) 849-29-34</p></bio><email xlink:type="simple">rodicvelic@gmail.com</email><xref ref-type="aff" rid="aff-1"/></contrib><contrib contrib-type="author" corresp="yes"><contrib-id contrib-id-type="orcid">https://orcid.org/0000-0002-9075-218X</contrib-id><name-alternatives><name name-style="eastern" xml:lang="ru"><surname>Вавилова</surname><given-names>Ю. Д.</given-names></name><name name-style="western" xml:lang="en"><surname>Vavilova</surname><given-names>J. D.</given-names></name></name-alternatives><bio xml:lang="ru"><p>Вавилова Юлия Дмитриевна — младший научный сотрудник.</p><p>Москва</p></bio><bio xml:lang="en"><p>Julia D. Vavilova - Junior Research Associate, Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences.</p><p>Moscow</p></bio><email xlink:type="simple">Juliateterina12@gmail.com</email><xref ref-type="aff" rid="aff-1"/></contrib><contrib contrib-type="author" corresp="yes"><contrib-id contrib-id-type="orcid">https://orcid.org/0000-0002-8996-2905</contrib-id><name-alternatives><name name-style="eastern" xml:lang="ru"><surname>Бойко</surname><given-names>А. А.</given-names></name><name name-style="western" xml:lang="en"><surname>Boyko</surname><given-names>A. A.</given-names></name></name-alternatives><bio xml:lang="ru"><p>Бойко Анна Александровна — кандидат биологических наук, научный сотрудник.</p><p>Москва</p></bio><bio xml:lang="en"><p>Anna A. Boyko - PhD (biology), Research Associate, Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences.</p><p>Moscow</p></bio><email xlink:type="simple">boyko_anna@mail.ru</email><xref ref-type="aff" rid="aff-1"/></contrib><contrib contrib-type="author" corresp="yes"><name-alternatives><name name-style="eastern" xml:lang="ru"><surname>Шустова</surname><given-names>О. A.</given-names></name><name name-style="western" xml:lang="en"><surname>Shustova</surname><given-names>O. A.</given-names></name></name-alternatives><bio xml:lang="ru"><p>Шустова Ольга Александровна — младший научный сотрудник.</p><p>Москва</p></bio><bio xml:lang="en"><p>Olga A. Shustova - Junior Research Associate, Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences.</p><p>Moscow</p></bio><email xlink:type="simple">shustolga.shusha@gmail.com</email><xref ref-type="aff" rid="aff-1"/></contrib><contrib contrib-type="author" corresp="yes"><contrib-id contrib-id-type="orcid">https://orcid.org/0000-0002-4470-4240</contrib-id><name-alternatives><name name-style="eastern" xml:lang="ru"><surname>Паламарчук</surname><given-names>А. И.</given-names></name><name name-style="western" xml:lang="en"><surname>Palamarchuk</surname><given-names>A. I.</given-names></name></name-alternatives><bio xml:lang="ru"><p>Паламарчук Анастасия Игоревна — аспирант, младший научный сотрудник.</p><p>Москва</p></bio><bio xml:lang="en"><p>Anastasia I. Palamarchuk - Postgraduate Student, Junior Research Associate, Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences.</p><p>Moscow</p></bio><email xlink:type="simple">palanastasia@yandex.ru</email><xref ref-type="aff" rid="aff-1"/></contrib><contrib contrib-type="author" corresp="yes"><contrib-id contrib-id-type="orcid">https://orcid.org/0000-0003-3056-4889</contrib-id><name-alternatives><name name-style="eastern" xml:lang="ru"><surname>Юсубалиева</surname><given-names>Г. М.</given-names></name><name name-style="western" xml:lang="en"><surname>Yusubalieva</surname><given-names>G. M.</given-names></name></name-alternatives><bio xml:lang="ru"><p>Юсубалиева Гаухар Маратовна — кандидат медицинских наук, старший научный сотрудник ФГБУ «ФНКЦ специализированных видов медицинской помощи и медицинских технологий» ФМБА России; старший научный сотрудник ФГБУН «ИМБ им. В.А. Энгельгардта» РАН; старший научный сотрудник ФГБУ «ФЦ мозга и нейротехнологий» ФМБА России.</p><p>Москва</p></bio><bio xml:lang="en"><p>Gaukhar M. Yusubalieva - PhD (Medicine), Senior Research Associate, Federal Research and Clinical Center of Specialized Medical Care and Medical Technologies, FMBA of Russia; Senior Research Associate, Engelhardt Institute of Molecular Biology of the Russian Academy of Sciences; Senior Research Associate, Federal Center of Brain Research and Neurotechnologies, FMBA of Russia.</p><p>Moscow</p></bio><email xlink:type="simple">gauhar@gauhar.org</email><xref ref-type="aff" rid="aff-2"/></contrib><contrib contrib-type="author" corresp="yes"><contrib-id contrib-id-type="orcid">https://orcid.org/0000-0003-3773-9629</contrib-id><name-alternatives><name name-style="eastern" xml:lang="ru"><surname>Кучерова</surname><given-names>О. Н.</given-names></name><name name-style="western" xml:lang="en"><surname>Kucherova</surname><given-names>O. N.</given-names></name></name-alternatives><bio xml:lang="ru"><p>Кучерова Ольга Николаевна — кандидат медицинских наук, заведующая отделением.</p><p>Москва</p></bio><bio xml:lang="en"><p>Olga N. Kucherova - PhD (Medicine), Head of Department, V. Vinogradov City Clinical Hospital, Department of Health of the City of Moscow, Branch 1 “Maternity Hospital 4”.</p><p>Moscow</p></bio><email xlink:type="simple">ola-kucherova@mail.ru</email><xref ref-type="aff" rid="aff-3"/></contrib><contrib contrib-type="author" corresp="yes"><contrib-id contrib-id-type="orcid">https://orcid.org/0000-0002-5403-0753</contrib-id><name-alternatives><name name-style="eastern" xml:lang="ru"><surname>Стрельцова</surname><given-names>М. А.</given-names></name><name name-style="western" xml:lang="en"><surname>Streltsova</surname><given-names>M. A.</given-names></name></name-alternatives><bio xml:lang="ru"><p>Стрельцова Мария Алексеевна — кандидат биологических наук, научный сотрудник.</p><p>Москва</p></bio><bio xml:lang="en"><p>Maria A. Streltsova - PhD (Biology), Research Associate, Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences.</p><p>Moscow</p></bio><email xlink:type="simple">mstreltsova@mail.com</email><xref ref-type="aff" rid="aff-1"/></contrib><contrib contrib-type="author" corresp="yes"><contrib-id contrib-id-type="orcid">https://orcid.org/0000-0001-8119-8247</contrib-id><name-alternatives><name name-style="eastern" xml:lang="ru"><surname>Коваленко</surname><given-names>Е. И.</given-names></name><name name-style="western" xml:lang="en"><surname>Kovalenko</surname><given-names>E. I.</given-names></name></name-alternatives><bio xml:lang="ru"><p>Коваленко Елена Ивановна — кандидат биологических наук, старший научный сотрудник.</p><p>Москва</p></bio><bio xml:lang="en"><p>Elena I. Kovalenko - PhD (Biology), Senior Research Associate, Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences.</p><p>Moscow</p></bio><email xlink:type="simple">lenkovalen@mail.ru</email><xref ref-type="aff" rid="aff-1"/></contrib></contrib-group><aff-alternatives id="aff-1"><aff xml:lang="ru"><institution>ФГБУН «Институт биоорганической химии имени академиков М.М. Шемякина и Ю.А. Овчинникова» Российской академии наук</institution><country>Россия</country></aff><aff xml:lang="en"><institution>Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences</institution><country>Russian Federation</country></aff></aff-alternatives><aff-alternatives id="aff-2"><aff xml:lang="ru"><institution>ФГБУ «Федеральный научно-клинический центр специализированных видов медицинской помощи и медицинских технологий» ФМБА России; ФГБУН «Институт молекулярной биологии имени В.А. Энгельгардта» Российской академии наук; ФГБУ «Федеральный центр мозга и нейротехнологий» ФМБА России</institution><country>Россия</country></aff><aff xml:lang="en"><institution>Federal Research and Clinical Center of Specialized Medical Care and Medical Technologies, FMBA of Russia; Engelhardt Institute of Molecular Biology of the Russian Academy of Sciences; Federal Center of Brain Research and Neurotechnologies, FMBA of Russia</institution><country>Russian Federation</country></aff></aff-alternatives><aff-alternatives id="aff-3"><aff xml:lang="ru"><institution>Городская клиническая больница имени В.В. Виноградова Департамента здравоохранения города Москвы, филиал 1 «Родильный дом 4»</institution><country>Россия</country></aff><aff xml:lang="en"><institution>V. Vinogradov City Clinical Hospital, Department of Health of the City of Moscow, Branch 1 “Maternity Hospital 4”</institution><country>Russian Federation</country></aff></aff-alternatives><pub-date pub-type="collection"><year>2023</year></pub-date><pub-date pub-type="epub"><day>01</day><month>06</month><year>2023</year></pub-date><volume>25</volume><issue>5</issue><fpage>1259</fpage><lpage>1264</lpage><permissions><copyright-statement>Copyright &amp;#x00A9; Величинский Р.А., Вавилова Ю.Д., Бойко А.А., Шустова О.A., Паламарчук А.И., Юсубалиева Г.М., Кучерова О.Н., Стрельцова М.А., Коваленко Е.И., 2023</copyright-statement><copyright-year>2023</copyright-year><copyright-holder xml:lang="ru">Величинский Р.А., Вавилова Ю.Д., Бойко А.А., Шустова О.A., Паламарчук А.И., Юсубалиева Г.М., Кучерова О.Н., Стрельцова М.А., Коваленко Е.И.</copyright-holder><copyright-holder xml:lang="en">Velichinskii R.A., Vavilova J.D., Boyko A.A., Shustova O.A., Palamarchuk A.I., Yusubalieva G.M., Kucherova O.N., Streltsova M.A., Kovalenko E.I.</copyright-holder><license xml:lang="ru" license-type="creative-commons-attribution" xlink:href="https://creativecommons.org/licenses/by/4.0/" xlink:type="simple"><license-p>Данная работа распространяется под лицензией Creative Commons Attribution 4.0.</license-p></license><license xml:lang="en" license-type="creative-commons-attribution" xlink:href="https://creativecommons.org/licenses/by/4.0/" xlink:type="simple"><license-p>This work is licensed under a Creative Commons Attribution 4.0 License.</license-p></license></permissions><self-uri xlink:href="https://www.mimmun.ru/mimmun/article/view/2846">https://www.mimmun.ru/mimmun/article/view/2846</self-uri><abstract><p>В настоящее время большое количество исследований по генной модификации NK-клеток пуповинной крови (UCB-NK) проводится как на клиническом, так и доклиническом уровне. Иммунотерапия на основе UCB-NK-клеток обладает большим терапевтическим потенциалом для использования в противоопухолевой терапии. Однако, несмотря на известные преимущества перед PB-NK-клетками, такие как высокая концентрация в пуповинной крови, низкий процент передачи вируса от донора, а также возможность сохранения фенотипа после криоконсервации, UCB-NK-клетки преимущественно характеризуют в научной литературе как незрелые и низкофункциональные NK-клетки. В данной работе были изучены фенотипические характеристики UCB-NK-клеток и возможность стимуляционной компенсации сниженной функциональной активности UCB-NK-клеток. Проведенные исследования выявили, что фенотипически UCB-NK-клетки можно охарактеризовать как малодифференцированные и слабоактивированные клетки, экспрессирующие высокий уровень ингибирующего рецептора NKG2A, низкий уровень активирующего рецептора NKG2C и молекулы активации HLA-DR, что соответствовало литературным данным. Для стимуляции свежевыделенных UCB-NK-клеток было выбрано два вида стимулов: 1) 100 ед IL-2; 2) комбинация 100 ед IL-2 и фидерных клеток K-562, экспрессирующих мембраносвязанный IL-21 (K562-mbIL21). Было показано, что при стимуляции UCB-NK-клеток в течение 7 дней комбинацией IL-2 и K562-mbIL21 уровень дегрануляции (LAMP-1) и пролиферативная активность этих клеток была выше, чем у параллельно культируемых в тех же условиях Ex vivo PB-NK-клеток, при этом стимул в виде IL-2 и K562-mbIL21 оказался более перспективным способом получения большого количества пролиферативно активных UCB-NK-клеток, по сравнению со стимуляцией только IL-2. Поскольку генетическая модификация NK-клеток является перспективным направлением улучшения противоопухолевых свойств NK-клеток, для дальнейшего изучения полученных UCB-NK-клеток была проведена процедура ретровирусной трансдукции. UCB-NK-клетки, стимулированные комбинацией IL-2 и K562-mbIL21, трансдуцировались на 8-й день культивирования. В данной работе применялась направленная оверэкспрессия адапторной молекулы DAP12, участвующей в сигналинге активирующих NK-клеточных рецепторов. PB-NK-клетки и UCB-NK-клетки трансдуцировали параллельно, в одинаковых экспериментальных условиях при равном объеме вирусных частиц. В результате было выявлено, что эффективность трансдукции вирусными частицами, несущими ген адапторной молекулы DAP12, в более чем 4 раза выше для UCB-NK-клеток по сравнению с PB-NK-клетками. Таким образом UCB-NK-клетки представляются перспективным инструментом для дальнейших исследований в области иммунотерапии рака.</p></abstract><trans-abstract xml:lang="en"><p>Currently, a large number of studies on genetic modification of cord blood NK cells (UCB-NK) are carried out at both clinical and preclinical levels. Immunotherapy based on UCB-NK cells has great potential for antitumor therapy. However, despite having known several advantages over peripheral blood NK cells (PB- NK), including a high concentration in cord blood and low virulence rate, UCB-NK cells are predominantly characterized in the scientific literature as immature and low-functioning NK cells. In this work, we studied the phenotypic characteristics of UCB-NK cells and the possibility of stimulatory compensation of the decreased functional activity of UCB-NK cells. Our studies revealed UCB-NK cells can be characterized as poorly differentiated and weakly activated cells with high level of inhibitory receptor NKG2A and low level of activating receptor NKG2C and HLA-DR, accordingly with the literature data. Two types of stimuli were chosen to stimulate freshly isolated UCB-NK cells: 1) 100 units of IL-2; 2) combinations of 100 units IL-2 and K-562 feeder cells expressing membrane-bound IL-21 (K562-mbIL21). It was shown the degranulation (LAMP-1) and proliferative activity was higher than for parallel cultured ex vivo PB-NK cells under the same conditions for UCB-NK cells stimulated for 7 days with IL-2 + K562-mbIL21. Moreover, stimulation in the way of IL-2 + K562-mbIL21 seemed to be a more perspective way to obtain a large number of proliferatively active UCB-NK cells compared to stimulation with IL-2 only. Since genetic modification of NK cells is a promising way to improve the antitumor properties of NK cells, retroviral transduction procedure was performed to study of the stimulated UCB-NK cells. UCB-NK cells stimulated with IL-2 + K562-mbIL21 were transduced on day 8 of cultivation. In this study, we used targeted overexpression of the adaptor molecule DAP12, which is involved in the signaling of activating NK cell receptors. PB-NK cells and UCB-NK cells were transduced under the equal experimental conditions in same volume of viral particles. As a result, the transduction efficiency was found to be more than 4-fold higher for UCB-NK cells compared to PB-NK cells. Thus, UCB-NK cells appear to be a promising tool for further research in cancer immunotherapy.</p></trans-abstract><kwd-group xml:lang="ru"><kwd>UCB-NK-клетки</kwd><kwd>фенотип</kwd><kwd>стимуляция NK-клеток</kwd><kwd>функциональная активность</kwd><kwd>трансдукция</kwd><kwd>DAP12</kwd></kwd-group><kwd-group xml:lang="en"><kwd>UCB-NK cells</kwd><kwd>phenotype</kwd><kwd>NK cell stimulation</kwd><kwd>functional activity</kwd><kwd>transduction</kwd><kwd>DAP12</kwd></kwd-group><funding-group><funding-statement xml:lang="ru">Исследование было поддержано грантом Российского научного фонда № 22-15-00503. Получение и выделение UCB-NK клеток было поддержано грантом Российского научного фонда № 22-64-00057.</funding-statement><funding-statement xml:lang="en">The study was supported by the Russian Science Foundation grant # 22-15-00503. 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