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目的:探讨紫外灭活型CVB3病毒诱导BALB/c小鼠产生特异性免疫应答及保护作用的评估。方法:采用紫外灭活的方法处理野生型CVB3m株,按照0.1 LD50(10~4 PFU)的剂量免疫小鼠,设置PBS免疫组作为对照,免疫后第3,5,7天收集小鼠血清,检测细胞因子含量;在第3,5,7,14天分离小鼠脾脏,流式分析T细胞亚群的分布比例;在免疫后一个月分离小鼠血清,检测中和抗体的滴度;同时间给予小鼠100LD 50野生型CVB3感染,观察小鼠的死亡率。结果:与对照组相比,在检测日期内紫外灭活型CVB3组小鼠血清中细胞因子IL-1α,TNF-α,IL-6的表达量明显增高(P<0.05),IL-4的表达量没有明显差异;免疫后第14天CD3~+CD4~+T细胞的分布较对照组明显升高(P<0.05);在免疫后一个月,紫外灭活型CVB3免疫组可以诱导机体产生高滴度中和抗体,同时,小鼠应对高致死量CVB3感染时有较高的存活率。结论:紫外灭活型CVB3感染能诱导机体产生特异性免疫应答,同时,产生的中和抗体可以提高小鼠应对致死剂量CVB3感染时的生存率,对机体有明显的保护作用。
OBJECTIVE: To evaluate the specific immune response induced by UV-inactivated CVB3 in BALB / c mice and its protective effect. Methods: Wild-type CVB3m strain was treated by UV inactivation. Mice were immunized with 0.1 LD50 (10 ~ 4 PFU), PBS immunized group was set as control, serum of mice were collected on days 3, 5 and 7 after immunization. The cytokines were detected; the spleens of mice were isolated on the 3rd, 5th, 7th and 14th days, and the distribution of T lymphocyte subsets was analyzed by flow cytometry. The serum of the mice was isolated one month after the immunization to detect the titer of the neutralizing antibody. Mice were challenged with 100 LD50 wild-type CVB3 at the time point to observe the mortality of mice. Results: Compared with the control group, the serum levels of IL-1α, TNF-α and IL-6 in UV-inactivated CVB3 group were significantly increased (P <0.05) The expression of CD3 ~ + CD4 ~ + T cells on the 14th day after immunization was significantly higher than that of the control group (P <0.05). One month after immunization, the UVB-inactivated CVB3 immunized group could induce the body to produce High titers of neutralizing antibodies, while mice should have a higher survival rate of CVB3 infection with high lethal dose. CONCLUSION: UVB-inoculated CVB3 can induce specific immune response in vivo. At the same time, the neutralizing antibody can improve the survival rate of mice when treated with lethal dose of CVB3 and has obvious protective effect on the body.