Study on immunological functions of heat shock protein 40 kDa of Schistosoma japonicum
10.16250/j.32.1374.2019175
- VernacularTitle:日本血吸虫热休克蛋白 40 kDa 免疫学功能研究
- Author:
Yan-Xiong YU
1
;
Sha-Sha LI
1
;
Ji-Feng ZHU
1
;
Xiao-Jun CHEN
1
;
Zhi-Peng XU
1
;
Ya-Lin LI
1
;
Sha ZHOU
1
;
Chuan SU
1
Author Information
1. Department of Pathogenic Biology, Nanjing Medical University, Nanjing 210029, China
- Publication Type:Journal Article
- Keywords:
Schistosoma japonicum;
Heat shock protein 40 kDa;
Immunological function;
Humoral immunity
- From:
Chinese Journal of Schistosomiasis Control
2020;32(3):262-267
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the immunological functions of heat shock protein 40 kDa of Schistosoma japonicum (SjHSP40). Methods The homology of the SjHSP40 protein sequence was analyzed and the B and T cell epitopes of SjHSP40 were predicted using bioinformatics tools. The full-length SjHSP40 gene was amplified using a PCR assay, and cloned into the prokaryotic expression vector pGEX-6P-1, which was transformed into Escherichia coli BL-21. The protein expression was induced with isopropyl β-D-thiogalactoside (IPDG), and then, the recombinant protein was purified with glutathione-sepharose 4B resin to yield the fusion protein GST-SjHSP40, which was checked with SDS-PAGE and Western blotting. Following immunization with GST-SjHSP40, the serum levels of anti-SjHSP40 IgG antibody and IgG1 and IgG2a subtypes were detected in BALB/c mice using ELISA. In addition, the effect of SjHSP40 on CD4+ T-cell subset differentiation was examined using flow cytometry. Results SjHSP40 contained 7 potential B cell epitopes and multiple T cell epitopes (CTL epitopes and Th epitopes). The prokaryotic expression plasmid pGEX-6p-1-SjSHP40 was successfully constructed, and the fusion protein GST-SjHSP40 was obtained following IPDG induction and protein purification. Significantly higher serum levels of anti-SjHSP40 IgG, IgG1 and IgG2a antibodies were detected in mice immunized with GST-SjHSP40 than in other groups; however, SjHSP40 showed no remarkable effects on CD4+ T-cell subset differentiation. Conclusions SjHSP40 may induce specific humoral immune responses in mice; however, it does not affect the balance of Th immune responses. It is suggested that SjHSP40 may be a potential vaccine candidate.
