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重组戊型肝炎病毒病毒样颗粒体外生物学活性研究

Biological activity of recombinant hepatitis E virus virus-like particles in vitro
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摘要 目的 评价重组戊型肝炎病毒(hepatitis E virus, HEV)病毒样颗粒(virus-like particle, VLP)体外生物学活性,为重组HEV VLP疫苗的应用提供基础研究数据。方法 将0.00(空白对照)、0.25、1.00、4.00μg/mL的重组HEV VLP分别与BALB/c小鼠的骨髓细胞和脾脏淋巴细胞共同培养。检测小鼠骨髓细胞MHCII、CD40和CD86分子的表达情况,评价重组HEV VLP促进小鼠骨髓细胞向抗原提呈细胞(antigen presenting cell, APC)分化和成熟的情况;分析脾脏淋巴细胞中活化的B淋巴细胞(CD19~+CD69~+)、活化的T淋巴细胞(CD3~+CD69~+)、辅助性T淋巴细胞(T helper lymphocytes, Th)(CD3~+CD4~+)和细胞毒性T淋巴细胞(cytotoxic T lymphocytes, CTL)(CD3~+CD8~+)细胞的比例,评价重组HEV VLP促进小鼠脾脏B淋巴细胞和T淋巴细胞活化和分化情况。结果 0.25、1.00、4.00μg/mL的重组HEV VLP刺激的小鼠骨髓细胞MHCII、CD40和CD86分子表达均升高,差异均有统计学意义(P均<0.05)。0.25、1.00、4.00μg/mL的重组HEV VLP刺激的小鼠脾脏淋巴细胞中活化的B淋巴细胞和T淋巴细胞比例均升高,差异均有统计学意义(P均<0.01)。1.00、4.00μg/mL的重组HEV VLP刺激的小鼠脾脏淋巴细胞中Th细胞比例均升高,差异均有统计学意义(P均<0.05)。结论 重组HEV VLP在小鼠体外实验中表现出良好的生物学活性,为重组HEV VLP疫苗的开发奠定了良好的理论和实验基础。 Objective To evaluate the in vitro biological activity of recombinant hepatitis E virus(HEV) virus-like particle(VLP),and provide basic research data for the application of VLP vaccine of HEV. Methods Bone marrow cells and spleen lymphocytes of BALB/c mice were isolated, and co-cultured with 0.25,1.00 and 4.00 μg/mL recombinant VLP of HEV,respectively, and a blank control(0.00 μg/mL) was set up. The expression of MHCII,CD40 and CD86 molecules was detected to evaluate the effect of VLP of recombinant HEV on the differentiation and maturation of antigen-presenting cells(APC). To evaluate the effect of VLP of recombinant HEV on the activation and differentiation of spleen B and T lymphocytes, the proportions of activated B lymphocytes(CD19~+CD69~+),activated T lymphocytes(CD3~+CD69~+),T helper lymphocytes(Th)(CD3~+CD4~+) and Cytotoxic T lymphocytes(CTL)(CD3~+CD8~+) of spleen lymphocytes were analyzed. Results The expressions of MHCII,CD40 and CD86 on mouse bone marrow cells stimulated by 0.25,1.00 and 4.00 μg/mL VLP of recombinant HEV were significantly increased(all P<0.05),indicating that VLP of recombinant HEV promoted the differentiation and maturation of mouse bone marrow cells into APC. The proportion of activated B lymphocytes and T lymphocytes in spleen lymphocytes stimulated by 0.25,1.00 and 4.00 μg/mL VLP of recombinant HEV was significantly increased(all P<0.01),meanwhile, the proportion of Th cells in spleen lymphocytes stimulated by 1.00 and 4.00 μg/mL VLP of recombinant HEV was significantly increased(all P<0.05),indicating that VLP of recombinant HEV promoted the differentiation and maturation of spleen lymphocytes in mice. Conclusion The VLP of recombinant HEV exhibited promising biological activity in vitro,which laid a solid theoretical and experimental foundation for the development of recombinant HEV VLP vaccines.
作者 秦海艳 谢忆 张巧玲 杨林鹏 刘宇阳 杨俊杰 QIN Haiyan;XIE Yi;ZHANG Qiaoling;YANG Linpeng;LIU Yuyang;YANG Junjie(The Four Research Department,Lanzhou Institute of Biological Products Co.,Ltd.,Center for Gansu Provincial Vaccine Engineering Research,Lanzhou 730046,Gansu Province,China)
出处 《微生物学免疫学进展》 CAS 2024年第2期14-20,共7页 Progress In Microbiology and Immunology
关键词 戊型肝炎病毒 病毒样颗粒 骨髓 脾脏 淋巴细胞 生物学活性 Hepatitis E virus(HEV) Virus-like particle(VLP) Bone marrow Spleen Lymphocyte Biological activity
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