布鲁氏菌Ⅳ型分泌系统对树突状细胞自噬、胞内生存和炎症反应的影响

doi:10.16431/j.cnki.1671-7236.2021.01.028

Abstract

Abstract: In order to explore the role of the type Ⅳ secretion system in the process of Brucella infection,in-depth understanding of the potential of the type Ⅳ secretion system of Brucella in vaccine development,in this study,the A19 vaccine strain of Brucella abortus was used as the research object,and the A19 VirB promoter deletion strain was used to infect mouse dendritic cells (DCs),evaluate the effect of type Ⅳ secretion system on the adhesion,invasion and intracellular survival of Brucella by colony count (CFU).At the same time,RNA and total protein were extracted from the infected cells,and the transcription and expression of the autophagy gene Beclin-1 were detected by Real-time PCR and Western blotting respectively.The cell supernatant after infection was collected,and ELISA was used to detect the secretion levels of inflammatory factors IL-6 and IL-10.The adhesion and invasion results showed that the adhesion and invasion levels of Brucella VirB promoter deletion strain and parent strain A19 were not significantly different (P>0.05).The intracellular survival experiment found that the intracellular survival ability of the Brucella VirB promoter deletion strain was significantly lower than that of the parent strain A19 at 4 h after infection (P<0.05),and it was extremely significantly lower at 0,24 and 48 h after infection (P<0.01).The results of Real-time PCR and Western blotting showed that the level of Beclin-1 production was extremely significantly higher than that of the parent strain A19 (P<0.01).ELISA results showed that at 8 and 12 h after infection,the level of IL-6 stimulated by VirB promoter deletion strain was significantly higher than that of parental strain A19 (P<0.05).However,at 8 and 12 h after infection,the level of IL-10 stimulated by VirB promoter deletion strain was significantly lower than that of parental strain A19 (P<0.05),and was extremely significantly lower than that of parental strain A19 at 24 h after infection (P<0.01).This study preliminarily explored the biological role of Brucella type Ⅳ secretion system in the process of infection of DCs,and laid a theoretical foundation for the follow-up Brucella vaccine transformation research.

Key words: Brucella VirB promoter deletion strain; type Ⅳ secretion system; DCs; intracellular survival; cytokines; autophagy

CLC Number:

S852.61

YANG Qin, DENG Xiaoyu, ZHAO Xiaoli, ZHANG Huan, XI Jing, YI Jihai, WANG Yong, ZHANG Qian, WANG Zhen, CHEN Chuangfu. Effects of Brucella Type Ⅳ Secretion System on Autophagy,Intracellular Survival and Inflammatory Response of Dendritic Cells[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(1): 257-264.

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Effects of Brucella Type Ⅳ Secretion System on Autophagy,Intracellular Survival and Inflammatory Response of Dendritic Cells

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