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基于沙眼衣原体主要外膜蛋白的多表位疫苗诱导小鼠产生特异性免疫。

A multi-epitope vaccine based on Chlamydia trachomatis major outer membrane protein induces specific immunity in mice.

机构信息

Department of Medical Microbiology and Immunology, Institute of Molecular Virology and Immunology, Wenzhou Medical University, Wenzhou 325000, China.

出版信息

Acta Biochim Biophys Sin (Shanghai). 2014 May;46(5):401-8. doi: 10.1093/abbs/gmu016. Epub 2014 Mar 28.

Abstract

We evaluated the immunogenicity and efficacy of a candidate vaccine comprising the major outer membrane protein (MOMP) multi-epitope of Chlamydia trachomatis. A short gene of multi-epitope derived from MOMP containing multiple T- and B-cell epitopes was artificially synthesized. The recombinant plasmid pET32a(+) containing codon optimized MOMP multi-epitope gene was constructed. Expression of the fusion protein Trx-His-MOMP multi-epitope in Escherichia coli was confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and western blot analysis. Balb/c mice were inoculated with the purified fusion protein subcutaneously three times with 2-week intervals. Results showed that the MOMP multi-epitope elicited not only strong humoral immune responses to C. trachomatis by generating significantly high levels of specific antibodies (IgG1 and IgG2a), but also a cellular immune response by inducing robust cytotoxic T lymphocyte responses in mice. Furthermore, the MOMP multi-epitope substantially primed secretion of IFN-γ, revealing that this vaccine could induce a strong Th1 response. Finally, the mice vaccinated with the MOMP multi-epitope displayed a reduction of C. trachomatis shedding upon a chlamydial challenge and an accelerated clearance of the infected C. trachomatis. In conclusion, the MOMP multi-epitope vaccine may have the potentiality for the development of effective prophylactic and therapeutic vaccines against the C. trachomatis infection.

摘要

我们评估了一种包含沙眼衣原体主要外膜蛋白(MOMP)多表位的候选疫苗的免疫原性和疗效。人工合成了含有多个 T 细胞和 B 细胞表位的 MOMP 多表位短基因。构建了含有密码子优化的 MOMP 多表位基因的重组质粒 pET32a(+)。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和 Western blot 分析证实了含有 Trx-His-MOMP 多表位融合蛋白的大肠杆菌表达。Balb/c 小鼠通过皮下接种三次纯化的融合蛋白,间隔 2 周。结果表明,MOMP 多表位不仅通过产生高水平的特异性抗体(IgG1 和 IgG2a)引起强烈的体液免疫反应,而且通过诱导小鼠强烈的细胞毒性 T 淋巴细胞反应引起细胞免疫反应。此外,MOMP 多表位大量引发 IFN-γ的分泌,表明该疫苗可以诱导强烈的 Th1 反应。最后,接种 MOMP 多表位的小鼠在衣原体感染后衣原体脱落减少,并加速清除感染的沙眼衣原体。总之,MOMP 多表位疫苗具有开发针对沙眼衣原体感染的有效预防性和治疗性疫苗的潜力。

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