CAJ | 학술논문

目的探讨含CpG基序的单链寡核苷酸(CpG ODN)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、CpG ODN预处理组和CpG ODN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、CpG ODN预处理组和CpG ODN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,CpG ODN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＜0.05).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.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淋巴细胞所占百分比、CD4+/CD8+和IFN-γ均升高(P＜0.05).结论 CpG ODN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.
목적 탐토함CpG기서적단련과핵감산(CpG ODN)간예대감염인륜상병독(HRV)유서세포면역응답적영향.방법 장40지ICR유서수모서안와수궤균분위대조조、HRV감염조、CpG ODN예처리조화CpG ODN치료조,매조10지.병독공격후제4천처사유서,무균취소장、비,대소장조직안통일손상표준평분,측정유서비장지수,새서람(MTT)비색법검측비림파세포자격지수,ELISA법검측비림파세포배양상청액중IFN-γ、IL-4적함량,류식세포의검측비T림파세포아군.채용단인소방차분석비교각조수거.결과 HRV감염조、CpG ODN예처리조화CpG ODN치료조장점막손상평분분별위4.00±1.31、2.75±1.28화2.87±0.99,각조간비교,차이유통계학의의(F=11.32,P＜0.01),여HRV감염조비교,CpG ODN예처리조、CpG ODN치료조적소장조직점막손상평분현저강저,차이유통계학의의(P＜0.05).여대조조비교,HRV감염조비림파세포자격지수、CD8+T림파세포소점백분비승고,CD4+T림파세포소점백분비、CD4+/CD8+강저,Th1세포인자IFN-γ표체증강,CpG ODN예처리조、CpG ODN치료조적비장지수、비장림파세포자격지수、CD4+T림파세포、CD8+T림파세포소점백분비、IFN-γ균승고(P＜0.05);여HRV감염조비교.CpG ODN예처리조、CpG ODN치료조적비장지수、비장림파세포자격지수、CD4+T림파세포소점백분비、CD4+/CD8+화IFN-γ균승고(P＜0.05).결론 CpG ODN증강감염HRV유서적세포면역응답,이Th1형세포면역응답위주.
Objective To investigate the effects of CpG oligodeoxynucleotide (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. Thedegree 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 cytometry. 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=ll. 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 IFN-y increased significantly in HRV infection group; while the spleen index, spleen lymphocyte stimulation index, percentages of CD4+ and CD8+ T lymphocytes, IFN-y 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 IFN-y 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 Th1 response.