| 刘辉萍,李舒怡,陈合意,陈超群.沙眼衣原体多表位疫苗的构建及其抗感染保护作用.[J].中南医学科学杂志.,2023,(2):183-187. |
| 沙眼衣原体多表位疫苗的构建及其抗感染保护作用 |
| Designing a multi-epitope vaccine candidate against Chlamydia trachomatis and evaluating its protective effect |
| 投稿时间：2022-08-12 修订日期：2023-02-25 |
| DOI：10.15972/j.cnki.43-1509/r.2023.02.007 |
| 中文关键词: 沙眼衣原体 生物信息学 多表位疫苗 抗感染保护 [ |
| 英文关键词:Chlamydia trachomatis bioinformatics multi-epitope vaccine anti-infection protection |
| 基金项目:湖南省教育厅创新平台开放基金项目(18K072)；南华大学大学生创新创业训练计划项目(220XCX511) |
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| 中文摘要: |
| 目的构建沙眼衣原体(Ct)多形态膜蛋白D(PmpD)与衣原体蛋白酶样活性因子(CPAF)多表位疫苗,并通过动物实验评价其抗感染保护作用。 方法利用生物信息学分析PmpD和CPAF的二级结构、抗原性、T细胞表位和B细胞表位；构建基于PmpD和CPAF多表位的重组蛋白RPC、原核表达、纯化RPC蛋白。免疫雌性BABL/c小鼠,ELISA检测小鼠血清IgG、IgG2a/IgG1及脾细胞上清细胞因子水平,CCK-8检测脾细胞增殖；Cm攻毒后,检测生殖道Cm载量以评估多表位融合疫苗RPC的抗感染保护作用。 结果成功构建多表位融合蛋白RPC。与PBS组相比,RPC组IgG抗体水平、IgG2a/IgG1比值、脾细胞刺激指数、γ干扰素、白细胞介素-2水平均明显升高(P＜0.05)；攻毒后RPC组可加快清除小鼠生殖道衣原体。 结论基于PmpD与CPAF的多表位融合蛋白RPC免疫小鼠,能诱导细胞免疫和体液免疫应答,有效降低衣原体载量,具有一定的抗感染保护作用。 |
| 英文摘要: |
| AimTo design a multi-epitope vaccine candidate against Chlamydia trachomatis (Ct) based on polymorphic membrane protein D (PmpD) and Chlamydial protease like active factor (CPAF), and to evaluate its anti-infection protection effect through animal tests. MethodsThe secondary structure, antigenicity, T- and B- cell epitopes of PmpD and CPAF were analyzed by bioinformatics techniques. Recombinant protein RPC was based on multiple epitopes of PmpD and CPAF was obtained by linking the dominant epitopes, and the RPC protein was expressed in prokaryotic expression system. After BABL/c female mice immunized with RPC protein, the levels of IgG and IgG2a/IgG1, the contents of cytokines in spleen cell supernatant were detected by ELISA. The proliferation of spleen cells was detected by CCK-8 assay. After challenge, the Chlamydia shedding of genital tract was measured to evaluate the anti-infection protective effect of multi-epitope fusion RPC. ResultsSuccessfully constructed multi-epitope fusion protein RPC. Compared with PBS group, the levels of IgG antibody, IgG2a/IgG1 ratio, spleen cell stimulation index, interferon-γ and interleukin-2 in RPC group were significantly higher (P＜0.05). After challenge, RPC group can accelerate the elimination of Chlamydia in reproductive tract of mice. ConclusionRPC, a multi-epitope vaccinecandidate based on Chlamydial PmpD and CPAF, could induce cellular and humoral immune responses in mice, had anti-infection protection and effectively reducing Chlamydia shedding. |
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