布鲁菌转录调节因子HFQ诱导机体免疫反应的分析

doi:10.11843/j.issn.0366-6964.2020.08.022

Abstract

Abstract: This experiment was conducted to study immune responses induced by Brucella transcriptional regulatory factor HFQ. In this study, a pair of primers were designed according to hfq gene sequence of B. abortus S2308 from GenBank (BAB1_1134), and the heat inactivated S2308 was used as template to amplify hfq gene by PCR. And then the hfq gene was cloned into pET-32a vector and transformed to E. coli BL21 (DE3) and induced the expression of recombinant HFQ protein (rHFQ). The expressed proteins were analyzed by SDS-PAGE and Western blot. Expression levels of cytokines IFN-γ and IL-4 in murine macrophages (RAW 264.7) stimulated with pET-32a empty plasmid, rHFQ and current vaccine M5-90 were detected by ELISA. Mice were immunized with pET-32a empty plasmid, rHFQ and current vaccine M5-90, and IFN-γ, IL-4 in splenocytes and IgG antibody levels in serum of mice were detected by ELISA. The results showed that the full length of hfq gene was 237 bp, encoding 79 amino acids. rHFQ was approximately 25.8 ku by the SDS-PAGE. There was a single band after purification. Results of Western blot indicated that the rHFQ had good reactogenicity. When RAW 264.7 cells were stimulated with pET-32a, rHFQ and M5-90, the levels of IFN-γ and IL-4 in rHFQ group were similar to those in M5-90 group, significantly higher than those in PBS group and pET-32a empty plasmid control group. The levels of IFN-γ and IL-4 were increased over time. When mice were immunized with pET-32a, rHFQ and M5-90, the levels of IFN-γ and IL-4 in splenocytes and IgG antibody levels in serum in rHFQ group were similar to those in M5-90 group, significantly higher than those in PBS group and pET-32a empty plasmid control group. The HFQ protein of Brucella has good reactivity and can induce the body to produce high levels of cellular and humoral immunity. It is an ideal candidate antigen for the development of Brucella subunit vaccines.

Key words: Brucella, HFQ, Immune responses

CLC Number:

WANG Shuli, ZHANG Huiru, BI Yanqi, WANG Dejuan, CHEN Lin, ZHANG Xiaoting, LI Zhiqiang. Analysis of Immune Responses Induced by Brucella Transcriptional Regulatory Factor HFQ[J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(8): 1977-1984.

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Analysis of Immune Responses Induced by Brucella Transcriptional Regulatory Factor HFQ

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