Левитес Е В, Кирикович С С, Долгова Е В, Проскурина А С, Риттер Г С, Останин А А, Черных Е Р, Богачев С С
Федеральный исследовательский центр Институт цитологии и генетики Сибирского отделения Российской академии наук, Новосибирск, Россия.
Федеральный исследовательский центр Институт цитологии и генетики Сибирского отделения Российской академии наук, Новосибирск, Россия Новосибирский национальный исследовательский государственный университет, Новосибирск, Россия.
Vavilovskii Zhurnal Genet Selektsii. 2020 May;24(3):284-291. doi: 10.18699/VJ20.621.
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.
本文报道了一种生产维生素D3结合蛋白（DBP）并将其转化为巨噬细胞激活因子GcMAF-RF的原创方法。根据原始方案，通过亲和色谱从人血浆中获得DBP，使用细胞固定化糖苷酶（β-半乳糖苷酶和神经氨酸酶）将其纯化并修饰为GcMAF-RF。使用特异性抗体通过蛋白质印迹法证实了血浆球蛋白Gc组中获得的多肽的存在。该多肽的分子特性使其与文献中描述的GcMAF蛋白相对应，该蛋白正在美国、英国、以色列和日本（在Saisei Mirai；里诺综合医疗中心；免疫生物技术有限公司；Efranat；以及催化长寿公司）进行临床试验。通过诱导巨噬细胞的吞噬活性及其在体外产生一氧化氮（NO）的能力来检测GcMAF-RF制剂的生物活性。通过巨噬细胞摄取磁珠的能力来评估其吞噬活性。通过捕获的磁珠与巨噬细胞总数的比率来计算巨噬细胞的激活程度。使用格里斯试剂通过比色法通过腹膜巨噬细胞培养上清液中一氧化氮的积累来估计NO的产生水平。结果表明，GcMAF-RF可使巨噬细胞的吞噬活性增加，并显著提高其一氧化氮的产生。巨噬细胞激活剂GcMAF-RF，根据其特性，与外国公司推向市场的类似制剂相对应，可被视为一种具有广泛作用谱的新型生物活性制剂。最令人感兴趣的是其通过激活巨噬细胞来增强适应性免疫的能力。在这方面，提出了GcMAF-RF的两个治疗用途领域。该制剂在癌症治疗领域将有需求，此外，它还可用于治疗多种神经退行性疾病。
