机构地区:[1]Department of Medical Microbiology and Immunology, Institute of Molecular Virology and Immunology, Wenzhou Medical University,Wenzhou 325000, China [2]Centre for Kidney Disease-Venomics Research, School of Medicine, The University of Queensland, Princess Alexandra Hospital,Brisbane QLD 4102, Australia
出 处:《Acta Biochimica et Biophysica Sinica》2014年第5期401-408,共8页生物化学与生物物理学报(英文版)
基 金:This work was supported in part by the grants from the National Natural Science Foundation of China (30972669), the Zhejiang Provincial Natural Science Foundation of China (Y2100611), and the Wenzhou Science and Technology Bureau (Y20100014, and Y20100090).
摘 要: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 muiti-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 multiepitope elicited not only strong humoral immune responses to C. trachomatis by generating significantly high levels of specific antibodies (lgG1 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 Thl 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.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 muiti-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 multiepitope elicited not only strong humoral immune responses to C. trachomatis by generating significantly high levels of specific antibodies (lgG1 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 Thl 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.
关 键 词:Chlamydia trachomatis VACCINE major outermembrane protein MULTI-EPITOPE humoral immuneresponse cytotoxic T lymphocyte (CTL) response
分 类 号:S852.5[农业科学—基础兽医学] Q93[农业科学—兽医学]
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