小反刍兽疫病毒疫苗毒株N75-1感染细胞源外泌体对山羊SLAM表达的影响

doi:10.11843/j.issn.0366-6964.2020.10.018

摘要/Abstract

摘要： 淋巴细胞信号活化分子（SLAM）是小反刍兽疫病毒（PPRV）在淋巴细胞的主要受体。本研究旨在探索PPRV感染细胞源外泌体对山羊SLAM表达的影响。通过荧光定量PCR法、流式细胞术和Western blot技术检测PPRV疫苗毒株N75-1感染山羊外周血单核细胞（PBMCs）源外泌体（Exo-PPRV）共育对接纳细胞（正常山羊PBMCs）中SLAM表达水平和细胞因子分泌水平的影响。结果表明，与正常细胞分泌外泌体（Exo-Mock）共育组相比，Exo-PPRV共培养组细胞SLAM mRNA和细胞表面表达水平均显著上调（P<0.05），TNF-α、IL-10和IFN-γ表达水平升高，而IFN-α水平降低（P<0.05）。进一步研究表明，Exo-PPRV中荷载高水平的PPRV H蛋白且可以将其传递至接纳细胞中，同时pcDNA3.1-H转染组中SLAM表达水平较pcDNA3.1对照转染组和未转染组明显升高。以上结果表明，PPRV感染PBMCs源外泌体对接纳细胞中SLAM表达水平具有显著正调控作用，外泌体中载荷高水平PPRV H蛋白是其调控接纳细胞SLAM表达的关键分子之一。

关键词: 外泌体, 小反刍兽疫病毒, 山羊PBMCs, 淋巴细胞信号活化分子

Abstract: Signaling lymphocyte activation molecule (SLAM) was identified as the primary receptor of Peste des petits ruminants virus (PPRV) on goat lymphocytes. This study aimed to explore the effects of exosomes derived from PPRV vaccine strain N75-1 infected cells on the expression of SLAM in goats. In this study, SLAM expression and cytokines expression levels of goat PBMCs (recipient cells) incubated with exosomes isolated from PPRV-infected goat PBMCs (Exo-PPRV) was analyzed by quantitative real-time PCR, flow cytometric assay and Western blot assay. The results showed that the recipient cells treated with Exo-PPRV had significantly increased SLAM mRNA and cell surface expression (P<0.05) as compared with Exo-Mock treated cells. Furthermore, the increased TNF-α, IL-10, and IFN-γ expression as well as decreased IFN-α expression (P<0.05) were also detected. Further study demonstrated that Exo-PPRV that contained high levels of PPRV H protein could transmit H protein to recipient cells. Moreover, the expression level of SLAM in the pcDNA3.1-H transfected cells was significantly higher than that in the pcDNA3.1 transfected cells and non-transfection group. Taken together, our study demonstrated that PPRV infected cells derived exosomes has positive effects on the SLAM expression in the recipient cells, and that the high level of PPRV H protein contained in PPRV-Exo is one of the key molecules for exosomes to regulate the expression of SLAM in the recipient cells.

Key words: exosomes, peste des petits ruminants virus, goat PBMCs, signaling lymphocyte activation molecule

中图分类号:

S852.659.5

褚云馨, 陈燕, 马雨晴, 曾巍, 陈松睿, 赵治钤, 王晶钰, 齐雪峰. 小反刍兽疫病毒疫苗毒株N75-1感染细胞源外泌体对山羊SLAM表达的影响[J]. 畜牧兽医学报, 2020, 51(10): 2500-2508.

CHU Yunxin, CHEN Yan, MA Yuqing, ZENG Wei, CHEN Songrui, ZHAO Zhiqian, WANG Jingyu, QI Xuefeng. Effect of Exosomes Derived from PPRV Vaccine Strain N75-1 Infected Cells on the Expression of SLAM in Goats[J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(10): 2500-2508.

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