Prohibitin 1/2 介导登革 3 型进入人神经母细胞瘤 (SH-SY5Y) 和小神经胶质细胞 (CHME-3) 细胞。
Prohibitin 1/2 mediates Dengue-3 entry into human neuroblastoma (SH-SY5Y) and microglia (CHME-3) cells.
机构信息
Department of Neurovirology, National Institute of Mental Health and Neurosciences, 560029, Bengaluru, India.
出版信息
J Biomed Sci. 2020 Apr 19;27(1):55. doi: 10.1186/s12929-020-00639-w.
BACKGROUND
Very few studies have identified receptor molecules for dengue virus (DENV) on neural cells. This study was designed to identify putative receptor/(s) involved in entry of DENV-3 in human neural cells of various lineages; neuronal-SH-SY5Y, astroglial-U-87 MG and microglial-CHME-3 cells.
RESULT
Virus overlay protein binding assay, LC-MS/MS and SEQUEST identified prohibitin1/2 (PHB1/2) as interacting proteins on SH-SY5Y, CHME-3, and U-87 MG cells. Infection inhibition and siRNA assays confirmed the role of PHB1/2 in the entry of DENV-3 into SH-SY5Y and CHME-3 cells but not in U-87 MG cells. Indirect immunofluorescence and flow-cytometry demonstrated the presence of PHB1/2 on the surface of SH-SY5Y and CHME-3 cells. Co-immunoprecipitation and Western blot, as well as double labelling, reconfirmed the interaction between PHB1/2 and DENV-3 EDIII protein.
CONCLUSION
These observations together for the first time indicate that PHB1/2 may serve as a putative receptor for DENV-3 in SH-SY5Y and CHME-3 cells. The study provided insights into DENV-3 and neural cell interactions.
背景
很少有研究鉴定出神经细胞上的登革热病毒（DENV）受体分子。本研究旨在鉴定参与 DENV-3 进入不同谱系的人神经细胞（神经元-SH-SY5Y、星形胶质细胞-U-87 MG 和小胶质细胞-CHME-3）的潜在受体/（s）。
结果
病毒覆盖蛋白结合分析、LC-MS/MS 和 SEQUEST 鉴定出 PHB1/2（PHB1/2）为 SH-SY5Y、CHME-3 和 U-87 MG 细胞上的相互作用蛋白。感染抑制和 siRNA 分析证实了 PHB1/2 在 DENV-3 进入 SH-SY5Y 和 CHME-3 细胞中的作用，但在 U-87 MG 细胞中不起作用。间接免疫荧光和流式细胞术表明 PHB1/2 存在于 SH-SY5Y 和 CHME-3 细胞的表面。免疫共沉淀和 Western blot 以及双标记进一步证实了 PHB1/2 与 DENV-3 EDIII 蛋白之间的相互作用。
结论
这些观察结果首次表明，PHB1/2 可能作为 DENV-3 在 SH-SY5Y 和 CHME-3 细胞中的潜在受体。该研究为 DENV-3 与神经细胞相互作用提供了新的见解。
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