伪狂犬病病毒疫苗株SA738侵染BHK-21细胞诱导其凋亡及Caspase3表达情况分析

Analysis of apoptosis and Caspase3 expression in BHK-21 cells infected by pseudorabies virus SA738 vaccine strain

旨在深入了解猪伪狂犬病病毒疫苗株PRV SA738侵染BHK-21细胞后,诱导细胞凋亡,细胞凋亡的时间及相关细胞凋亡因子表达情况。通过测定传统疫苗毒株Bartha-K61株和实验室人工筛选的gI-/gE-/PRV SA738病毒株滴度,分别利用2株疫苗毒株侵染BHK-21,采用原位双荧光染色法检测细胞凋亡,用分光光度法测定细胞凋亡因子Caspase-3表达情况。结果表明,低剂量gI-/gE-/PRV SA738疫苗毒株和Bartha-K61疫苗毒株均延迟侵染细胞的凋亡。缺失疫苗毒株gI-/gE-/PRV SA738延迟侵染细胞的凋亡时间比Bartha-K61疫苗毒株时间长,但侵染细胞增殖速度下降。细胞凋亡后期最终细胞死亡,正常细胞细胞死亡后无凋亡小体。gI-/gE-/PRV SA738疫苗毒株和Bartha-K61疫苗毒株侵染细胞凋亡后期凋亡小体形成,并检测到膜结构。在病毒侵染早期细胞内关键凋亡细胞因子Caspase3表现为上调。Caspase-3参与了病毒侵染过程中细胞凋亡的调控。

To future understand the institution of induced cell apoptosis and the expression of cell apoptosis factors after infection BHK-21 cell line with pseudorabies virus vaccine strain SA738. Through the determination of the virus titer of classical vaccine strain PRV Bartha-K61 and PRV gI-/gE-/PRV SA738 vaccine strain,used two vaccine strains infected BHK-21 cell line respective- ly,detected apoptotic ceils in situ by immunofluorescence staining and expression of cell apoptosis factor caspase-B with spectrophotometric assay. The results showed that tile low dose of SA738 gI /gE- / PRV strain and Bartha-K61 strain both delayed the infected cells apoptosis. The apopto- sis of SA738 gI-/gE-/PRV infected cells was longer than that of Bartha-K61 vaccine,but the cell proliferation rate was decreased, finally cell was up to death in the late stage, and no apoptotic bodies were found for the normal cell . however in the early stage of virus infection,the key apoptotic cell factor Caspase3 showed up-regulation after PRV infected cell and apoptotic bodies and the membrane structure was detected in the late stage. Caspase3 attended in the regulation of cell apoptosis in the process of PRV virus infection.