中国人兽共患病学报2025，Vol.41Issue(10)：999-1004,6.DOI:10.3969/j.issn.1002-2694.2025.00.165
结核分枝杆菌Rv2318及其表位肽免疫原性研究
Immunogenicity of Rv2318 and its epitope peptides of Mycobacterium tuberculosis
摘要
Abstract
To screen new antigens for novel tuberculosis(TB)vaccine research,we used bioinformatics to predict the B and T cell epitopes of Rv2318,and evaluated the immunogenicity of Rv2318 and its T/B epitope peptides(Rv2318p).The recombinant plas-mids pET32a-Rv2318 and pET32a-Rv2318p were constructed through gene synthesis methods.The recombinant proteins were ex-pressed in a prokaryotic system and purified with nickel affinity chromatography.Proteins were identified with SDS-PAGE and western blotting.BALB/c mice were immunized subcutaneously with the recombinant proteins to evaluate immunogenicity.Sera were collected,and antigen specific antibody titers were evaluated with ELISA.Splenocytes were isolated,and cytokines and T cell proliferation were analyzed with ELISA and flow cytometry,respectively.Rv2318 included two epitope fragments,aa10-130 and 350-410.SDS-PAGE and western blotting indicated that the target proteins were expressed and purified correctly,and their relative molecular weights were-approximately 68 kD and 42 kD,respectively.Rv2318 and Rv2318p induced stronger humoral immune responses than observed in the control groups(P<0.000 1,n=6).Compared with Rv2318,Rv2318p showed significantly greater enhancement of specific IgG and IgG subclass antibodies(P<0.000 1,n=6).In addition,Rv2318p increased the ratio of IgG2a/IgG1,thus indicating that it primarily induced a cellular immune response biased toward the Th1 type.Cytokine experiments revealed that IFN-γ,IL-2,IL-6,and IL-4 significantly increased after immunization with Rv2318p(P all<0.01,n=6),particularly Th1 type cytokines(IFN-γ and IL-2).Furthermore,Rv2318 increased the expression of only IL-2 and IL-6,particularly IL-6(P all<0.01,n=6).Although Rv2318 in-duced more IFN-γ,we observed no significant difference between Rv2318 and PBS immunized mice.Importantly,Rv2318p stimu-lated mice to express IFN-γ at 842 pg/mL,approximately 3 times the level elicited by Rv2318.Whereas both proteins increased the proportions of CD4+and CD8+T cells,Rv2318p promoted greater proliferation of T lymphocytes.These data indicated that both Rv2318 and its epitope peptides enhanced humoral and cellular immune responses,whereas the epitope peptides notably triggered a stronger Th1 type cellular response.In conclusion,the recombinant protein Rv2318 and its epitope peptides showed favorable immunogenicity,and the immunogenicity of Rv2318p was superior to that of Rv2318.This study provides a theoretical basis for TB vaccine development.关键词
结核分枝杆菌/Rv2318/表位肽/免疫原性Key words
Mycobacterium tuberculosis/Rv2318/epitope peptides/immunogenicity分类
医药卫生引用本文复制引用
范雪亭,赵秀芹,刘海灿,王瑞欢,李马超,万康林,赵丽丽,王瑞白,郭溢,李桂莲..结核分枝杆菌Rv2318及其表位肽免疫原性研究[J].中国人兽共患病学报,2025,41(10):999-1004,6.基金项目
国家重点研发计划(No.2023 YFC2307201) （No.2023 YFC2307201）
传染病防控能力提升青年基金项目(No.102393240020020000003) National Key R&D Program of China(No.2023 YFC2307201) （No.102393240020020000003）
Youth Fund for Enhancing Capability of Infectious Disease Surveillance and Prevention(No.102393240020020000003) （No.102393240020020000003）
