摘要
目的探讨含CpG基序的单链寡核苷酸（CpGODN）干预对感染人轮状病毒（HRV）乳鼠细胞免疫应答的影响。方法将40只ICR乳鼠随母鼠按窝随机均分为对照组、HRV感染组、CpGODN预处理组和CpOODN治疗组，每组10只。病毒攻击后第4天处死乳鼠，无菌取小肠、脾，对小肠组织按统一损伤标准评分，测定乳鼠脾脏指数，噻唑蓝（MTT）比色法检测脾淋巴细胞刺激指数，ELISA法检测脾淋巴细胞培养上清液中IFN-γ、IL-4的含量，流式细胞仪检测脾T淋巴细胞亚群。采用单因素方差分析比较各组数据。结果HRV感染组、CpGODN预处理组和CpGODN治疗组肠黏膜损伤评分分别为4．00±1．31、2．75±1．28和2．87±0．99，各组间比较，差异有统计学意义（F=11．32，P〈0．01），与HRV感染组比较，CF）GODN预处理组、CpG ODN治疗组的小肠组织黏膜损伤评分显著降低，差异有统计学意义（P〈O．05）。与对照组比较，HRV感染组脾淋巴细胞刺激指数、CD8＋T淋巴细胞所占百分比升高，CD4＋T淋巴细胞所占百分比、CD4＋／CD8＋降低，Th1细胞因子IFN-γ表达增强，CpG ODN预处理组、CpG ODN治疗组的脾脏指数、脾脏淋巴细胞刺激指数、CD4＋T淋巴细胞、CD8＋T淋巴细胞所占百分比、IFN-γ均升高（P〈0．05）；与HRV感染组比较，CpGODN预处理组、CpGODN治疗组的脾脏指数、脾脏淋巴细胞刺激指数、CD4＋T淋巴细胞所占百分比、CD4＋／CD8＋和IFN-γ均升高（P〈0．05）。结论CpGODN增强感染HRV乳鼠的细胞免疫应答，以Th1型细胞免疫应答为主。
Objective To investigate the effects of CpG oligodeoxynueleotide （ODN） on cellular immune responses in suckling mice infected with human rotavirus （HRV）. Methods Forty ICR suckling mice were randomly divided into four groups： control group, HRV infection group, CpG ODN pretreatment group and CpG ODN treatment group. Suckling mice were sacrificed four days after rotavirus challenge. Small intestine and spleen samples were collected under sterile condition. The degree of small intestinal mucosal injures was evaluated with standard scoring criteria. The spleen index was calculated and spleen lymphocyte stimulation index was detected by methyl thiazolyl tetrazolium （MTT） assays. Levels of gamma interferon （IFN-γ）, interleukin-4 （IL-4） in the supernatant of spleen lymphocyte culture were detected by enzyme linked immunosorbent assay （ELISA）. T lymphocyte subsets were analyzed by flow eytometry. The data was compared by one way ANOVA. Results The scores of mucosal injures of mice in HRV infection group, CpG ODN pretreatment group and CpG ODN treatment group were 4.00±1.31, 2.75 ± 1.28 and 2.87 ± 0.99, respectively, and the differences among groups were statistically significant （F = 11. 32, P〈0. 01）. The scores of mucosal injures in CpG ODN pretreatment group and CpG ODN treatment group were both lower than that in HRV infection group （P〈0. 05）. Compared with control group, spleen lymphocyte stimulation index and the percentage of CD8＋ T lymphocytes were higher, the percentage of CD4＋ T lymphocyte and ratio of CD4＋/CD8＋ T cells were lower, levels of T helper （Th）1 type cytokine IFN7 increased significantly in HRV infection group; while the spleen index, spleen lymphocyte stimulation index, percentages of CD4＋ and CD8＋ T lymphocytes, IFN-γ levels were all increased in CpG ODN pretreatment group and CpG ODN treatment group （P〈0. 05）. The spleen index, spleen lymphocyte stimulation index, the percentage of CD4＋ T lymphocytes, CD4＋/CD8＋ ratios and IFNy levels in CpG ODN pretreatment group and CpG ODN treatment group were all significantly higher than HRV infection group （P〈0.05）. Conclusion CpG ODN potently enhances cellular immune responses in ICR suckling mice infected with HRV and CpG ODN could induce dominant Thl response.
出处
《中华传染病杂志》
CAS
CSCD
北大核心
2011年第6期329-333,共5页
Chinese Journal of Infectious Diseases
基金
浙江省卫生厅基金资助项目（2010KYA139）
温州市科技局基金资助项目（Y20090350）
