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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">vavilov</journal-id><journal-title-group><journal-title xml:lang="ru">Вавиловский журнал генетики и селекции</journal-title><trans-title-group xml:lang="en"><trans-title>Vavilov Journal of Genetics and Breeding</trans-title></trans-title-group></journal-title-group><issn pub-type="epub">2500-3259</issn><publisher><publisher-name>Institute of Cytology and Genetics of Siberian Branch of the RAS</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="doi">10.18699/VJ20.621</article-id><article-id custom-type="elpub" pub-id-type="custom">vavilov-2596</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>ГЕНЕТИКА МИКРООРГАНИЗМОВ</subject></subj-group><subj-group subj-group-type="section-heading" xml:lang="en"><subject>MICROBIAL GENETICS</subject></subj-group></article-categories><title-group><article-title>Оценка in vitro биологической активности отечественного препарата макрофаг-активирующего фактора (GcMAF-RF)</article-title><trans-title-group xml:lang="en"><trans-title>In vitro assay of biological activity of a national preparation of macrophage activating factor (GcMAF-RF)</trans-title></trans-title-group></title-group><contrib-group><contrib contrib-type="author" corresp="yes"><contrib-id contrib-id-type="orcid">https://orcid.org/0000-0002-3093-407X</contrib-id><name-alternatives><name name-style="eastern" xml:lang="ru"><surname>Левитес</surname><given-names>Е. В.</given-names></name><name name-style="western" xml:lang="en"><surname>Levites</surname><given-names>E. V.</given-names></name></name-alternatives><bio xml:lang="ru"><p>Новосибирск</p></bio><bio xml:lang="en"><p>Novosibirsk</p></bio><xref ref-type="aff" rid="aff-1"/></contrib><contrib contrib-type="author" corresp="yes"><contrib-id contrib-id-type="orcid">https://orcid.org/0000-0002-3426-4501</contrib-id><name-alternatives><name name-style="eastern" xml:lang="ru"><surname>Кирикович</surname><given-names>С. С.</given-names></name><name name-style="western" xml:lang="en"><surname>Kirikovich</surname><given-names>S. S.</given-names></name></name-alternatives><bio xml:lang="ru"><p>Новосибирск</p></bio><bio xml:lang="en"><p>Novosibirsk</p></bio><xref ref-type="aff" rid="aff-1"/></contrib><contrib contrib-type="author" corresp="yes"><contrib-id contrib-id-type="orcid">https://orcid.org/0000-0002-5543-248X</contrib-id><name-alternatives><name name-style="eastern" xml:lang="ru"><surname>Долгова</surname><given-names>Е. В.</given-names></name><name name-style="western" xml:lang="en"><surname>Dolgova</surname><given-names>E. V.</given-names></name></name-alternatives><bio xml:lang="ru"><p>Новосибирск</p></bio><bio xml:lang="en"><p>Novosibirsk</p></bio><xref ref-type="aff" rid="aff-1"/></contrib><contrib contrib-type="author" corresp="yes"><contrib-id contrib-id-type="orcid">https://orcid.org/0000-0002-7650-4331</contrib-id><name-alternatives><name name-style="eastern" xml:lang="ru"><surname>Проскурина</surname><given-names>А. С.</given-names></name><name name-style="western" xml:lang="en"><surname>Proskurina</surname><given-names>A. S.</given-names></name></name-alternatives><bio xml:lang="ru"><p>Новосибирск</p></bio><bio xml:lang="en"><p>Novosibirsk</p></bio><xref ref-type="aff" rid="aff-1"/></contrib><contrib contrib-type="author" corresp="yes"><contrib-id contrib-id-type="orcid">https://orcid.org/0000-0003-1573-3795</contrib-id><name-alternatives><name name-style="eastern" xml:lang="ru"><surname>Риттер</surname><given-names>Г. С.</given-names></name><name name-style="western" xml:lang="en"><surname>Ritter</surname><given-names>G. S.</given-names></name></name-alternatives><bio xml:lang="ru"><p>Новосибирск</p></bio><bio xml:lang="en"><p>Novosibirsk</p></bio><xref ref-type="aff" rid="aff-2"/></contrib><contrib contrib-type="author" corresp="yes"><contrib-id contrib-id-type="orcid">https://orcid.org/0000-0001-6895-938X</contrib-id><name-alternatives><name name-style="eastern" xml:lang="ru"><surname>Останин</surname><given-names>А. А.</given-names></name><name name-style="western" xml:lang="en"><surname>Ostanin</surname><given-names>А. A.</given-names></name></name-alternatives><bio xml:lang="ru"><p>Новосибирск</p></bio><bio xml:lang="en"><p>Novosibirsk</p></bio><xref ref-type="aff" rid="aff-3"/></contrib><contrib contrib-type="author" corresp="yes"><contrib-id contrib-id-type="orcid">https://orcid.org/0000-0003-2346-6279</contrib-id><name-alternatives><name name-style="eastern" xml:lang="ru"><surname>Черных</surname><given-names>Е. Р.</given-names></name><name name-style="western" xml:lang="en"><surname>Chernykh</surname><given-names>E. R.</given-names></name></name-alternatives><bio xml:lang="ru"><p>Новосибирск</p></bio><bio xml:lang="en"><p>Novosibirsk</p></bio><xref ref-type="aff" rid="aff-3"/></contrib><contrib contrib-type="author" corresp="yes"><contrib-id contrib-id-type="orcid">https://orcid.org/0000-0002-2019-9382</contrib-id><name-alternatives><name name-style="eastern" xml:lang="ru"><surname>Богачев</surname><given-names>С. С.</given-names></name><name name-style="western" xml:lang="en"><surname>Bogachev</surname><given-names>S. S.</given-names></name></name-alternatives><bio xml:lang="ru"><p>Новосибирск</p></bio><bio xml:lang="en"><p>Novosibirsk</p></bio><email xlink:type="simple">labmolbiol@mail.ru</email><xref ref-type="aff" rid="aff-1"/></contrib></contrib-group><aff-alternatives id="aff-1"><aff xml:lang="ru">Федеральный исследовательский центр Институт цитологии и генетики, Сибирское отделение Российской академии наук<country>Россия</country></aff><aff xml:lang="en">Institute of Cytology and Genetics, Siberian Branch of the Russian Academy of Sciences<country>Russian Federation</country></aff></aff-alternatives><aff-alternatives id="aff-2"><aff xml:lang="ru">Федеральный исследовательский центр Институт цитологии и генетики, Сибирское отделение Российской академии наук; Новосибирский национальный исследовательский государственный университет<country>Россия</country></aff><aff xml:lang="en">Institute of Cytology and Genetics, Siberian Branch of the Russian Academy of Sciences; Novosibirsk State University<country>Russian Federation</country></aff></aff-alternatives><aff-alternatives id="aff-3"><aff xml:lang="ru">Научно-исследовательский институт фундаментальной и клинической иммунологии<country>Россия</country></aff><aff xml:lang="en">Institute of Fundamental and Clinical Immunology<country>Russian Federation</country></aff></aff-alternatives><pub-date pub-type="collection"><year>2020</year></pub-date><pub-date pub-type="epub"><day>29</day><month>05</month><year>2020</year></pub-date><volume>24</volume><issue>3</issue><fpage>284</fpage><lpage>291</lpage><permissions><copyright-statement>Copyright &amp;#x00A9; Левитес Е.В., Кирикович С.С., Долгова Е.В., Проскурина А.С., Риттер Г.С., Останин А.А., Черных Е.Р., Богачев С.С., 2020</copyright-statement><copyright-year>2020</copyright-year><copyright-holder xml:lang="ru">Левитес Е.В., Кирикович С.С., Долгова Е.В., Проскурина А.С., Риттер Г.С., Останин А.А., Черных Е.Р., Богачев С.С.</copyright-holder><copyright-holder xml:lang="en">Levites E.V., Kirikovich S.S., Dolgova E.V., Proskurina A.S., Ritter G.S., Ostanin А.A., Chernykh E.R., Bogachev S.S.</copyright-holder><license 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://vavilov.elpub.ru/jour/article/view/2596">https://vavilov.elpub.ru/jour/article/view/2596</self-uri><abstract><p>В статье сообщается о разработанном оригинальном способе получения витамин D3-связываю-щего белка (DBP) и его конвертации в макрофаг-активирующий фактор GcMAF-RF. Согласно разработанному регламенту, DBP получали из плазмы крови человека, применяя аффинную колоночную хроматографию, очищали и модифицировали до GcMAF-RF с использованием цитоиммобилизованных гликозидаз (бета-галакто-зидаза и нейраминидаза). Принадлежность полученного полипептида к Gc-группе глобулинов плазмы крови подтверждали вестерн-блотом с использованием специфических антител. Полученный полипептид по своим молекулярным свойствам соответствует описанному в литературе белку GсMAF, находящемуся на стадии клинических испытаний в США, Британии, Израиле и Японии (Saisei Mirai, Reno Integrative Medical Center, Immuno Biotech Ltd, Efranat, Catalytic Longevity). Биологическую активность препарата GcMAF-RF определяли по индукции у перитонеальных макрофагов мыши фагоцитарной активности и способности продуцировать монооксид азота (NO) in vitro. Фагоцитарную активность макрофагов оценивали по эффективности захвата магнитных шариков. Степень активации макрофагов рассчитывали по отношению числа захваченных шариков к общему числу макрофагов. Уровень продукции NO оценивали по накоплению монооксида азота в культуральных супернатантах перитонеальных макрофагов колориметрическим методом с использованием реактива Грисса. Показано, что GcMAF-RF кратно увеличивает фагоцитарную активность макрофагов и достоверно увеличивает продукцию ими монооксида азота. Выделенный оригинальным способом активатор макрофагов GcMAF-RF по своим характеристикам (согласно материалам, опубликованным в печати) соответствует препаратам GcMAF, представляемым на рынке зарубежными компаниями, и может рассматриваться как новый отечественный биологически активный препарат с широким спектром действия. Наибольший интерес вызывает его способность через активацию макрофагов усиливать адаптивный иммунитет организма. В этой связи предполагаются два направления терапевтического применения препарата GcMAF-RF. Препарат может быть востребован в области лечения онкологических заболеваний и, кроме того, может быть использован при лечении ряда нейродегенеративных патологий и иммунодефицитных состояний.</p></abstract><trans-abstract xml:lang="en"><p>The article reports an original method for producing vitamin D3-binding protein (DBP) and its conversion into macrophage-activating factor GcMAF-RF. According to an original protocol, DBPs were obtained from human blood plasma using affinity chromatography, purified and modified to GcMAF-RF using cytoimmobilized glycosidases (beta-galactosidase and neuraminidase). The presence of the polypeptide obtained in the Gc group of blood plasma globulins was confirmed by Western blot using specific antibodies. The molecular properties of this polypeptide put it in correspondence with the GcMAF protein described in the literature, which is undergoing clinical trials in the USA, Britain, Israel and Japan (at Saisei Mirai; Reno Integrative Medical Center; Immuno Biotech Ltd; Efranat; and Catalytic Longevity). The biological activity of the GcMAF-RF preparation was detected by the induction of phagocytic activity of macrophages and their ability to produce nitrogen monoxide (NO) in vitro.The phagocytic activity of macrophages was evaluated by their ability to uptake magnetic beads. The degree of activation of macrophages was calculated by the ratio of trapped beads to the total number of macrophages. The level of NO production was estimated by the accumulation of nitrogen monoxide in the culture supernatants of peritoneal macrophages by the colorimetric method using the Griess reagent. It was shown that GcMAF-RF multiplies the phagocytic activity of macrophages and significantly increases their production of nitrogen monoxide. The macrophage activator GcMAF-RF, according to its characteristics, corresponds to similar preparations which are made available to the market by foreign companies, and can be considered as a new biologically active preparation with a wide spectrum of action. Of greatest interest is its ability - through the activation of macrophages - to enhance the adaptive immunity. In this regard, two areas of therapeutic use of the GcMAF-RF are proposed. The preparation will be in demand in the field of cancer treatment, and, in addition, it can be used in the treatment of a number of neurodegenerative pathologies.</p></trans-abstract><kwd-group xml:lang="ru"><kwd>макрофаг-активирующий фактор (GcMAF)</kwd><kwd>витамин D3-связывающий белок (DBP)</kwd><kwd>фагоцитоз</kwd><kwd>монооксид азота (NO)</kwd><kwd>перитонеальные макрофаги</kwd></kwd-group><kwd-group xml:lang="en"><kwd>Gc protein-derived macrophage activating factor (GcMAF)</kwd><kwd>vitamin D3-binding protein (DBP)</kwd><kwd>phagocytosis</kwd><kwd>nitrogen monoxide (NO)</kwd><kwd>peritoneal macrophages</kwd></kwd-group><funding-group xml:lang="en"><funding-statement>The authors are grateful to Head of the Shared Access Center "Cell technologies” Institute of Cytology and Genetics, Novosibirsk, for access to the microscope. 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