重组2型登革病毒非结构1蛋白体外诱导外周血单核细胞TNF-α水平的研究

V. Kurnianda, S. Faradilla, S. Karina, S. Agustina, M. Ulfah, C. Octavina, F. Syahliza, M. R. Ramadhan, S. Purnawan, M. Musman
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引用次数: 0

摘要

登革热感染是一个全球卫生问题,发病率每年都在增加,目前在100多个世卫组织国家流行。登革病毒(DENV)是一种单链阳性RNA病毒,基因组大小为±11kb,编码3个结构蛋白、7个非结构蛋白和2个非翻译区(UTR)。已知NS1蛋白在DENV严重感染的发生发展中具有非常重要的作用,其直接作用是引起宿主细胞损伤,间接作用是激活免疫应答诱导过量细胞因子的分泌。用重组pcNS1质粒转染CHO-K1细胞与健康供体的PBMC共培养。共培养48h后,收集细胞上清,ELISA检测TNF-α水平和NS1重组蛋白。重组NS1蛋白在CHO-K1哺乳动物细胞系中表达,诱导TNF-α水平高于对照组。重组NS1蛋白表达能诱导TNF-α分泌,且分泌水平高于对照组。
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Levels of TNF-α in PBMC (Peripheral Blood Mononuclear Cells) Induced by Recombinant Non Structural 1 Protein of Dengue Virus Serotype-2 in vitro
Dengue infection is a global health problem with an increasing incidence every year and now endemic in more than 100 WHO countries. Dengue infection is caused by dengue virus (DENV) which is an RNA virus with positive single strand, with ±11kb genome size encoding 3 structural proteins, 7 non-structural proteins, and two Untranslated Region (UTR). NS1 protein is known to have a very important role in the development of severe DENV infection, by the direct effect causing host cells damage and indirect effect by activating immune response to induce the secretion of excess cytokines. Transfected CHO-K1 cells with recombinant pcNS1 plasmid was co-cultured with PBMC from healthy donor. 48 hours post co-cultured, cell supernatant was collected and TNF-α levels and NS1 recombinant were measured by ELISA. Recombinant NS1 protein was expressed in CHO-K1 mammalian cell line and able to induce TNF-α with higher levels compared to control. Recombinant NS1 protein expression was able to induce secretion of TNF-α with higher levels compared to control.
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