摘要
目的建立鼠巨细胞病毒（MCMV）播散性感染的小鼠模型，在整体水平观察脾脏病理损伤与促炎因子IL-17A表达的关系，初步探讨IL-17A参与巨细胞病毒感染后脾脏病理改变的机制。方法建立鼠巨细胞病毒全身播散性感染模型，MCMVSmith株腹腔接种后3、7、14、28d各处死小鼠3只，同时设正常小鼠作为模拟感染对照。标准空斑实验检测脾脏的病毒滴度，lit．PCR法测定脾脏中MCMVmRNA、IL-17AmRNA的表达，免疫组化法检测脾脏的IL-17A蛋白的表达，HE染色法评估脾脏的病理性损伤程度，分析IL-17A的表达与脾脏病理改变的关系。结果脾组织的病毒滴度在巨细胞病毒感染后3d升高，7d有所降低，在感染后第14天用标准空斑实验检测不到病毒。MCMVmRNA在病毒感染后3、7d可以检测的到；IL-17AmRNA的表达，与模拟感染对照组相比，MCMV感染组表达逐渐增加，并在14d达到高峰，28d显著下降。免疫组化法检测的脾脏IL-17A蛋白的表达也在14d达到高峰。脾组织的病理损伤逐渐加重，至感染后第14天病变达高峰后逐渐减轻。脾脏组织病理损伤最重与IL-17A高表达出现在同一时期。结论促炎因子IL-17A的高表达与脾脏组织的病理损伤程度呈现明显相关性。
Objective To explore the correlation between the expression of IL-17A and the degree of spleen damage in acute mouse cytomegalovirus（MCMV） disseminated infection in vivo and to understand the mechanism about how IL-17A involved in the pathological damage of the spleen in MCMV infection. Methods An acute disseminated MCMV infection model was established in mice. BALB/c mice were ran- domly divided into two groups. Mice in group one were infected with MCMV Smith to establish disseminated infection. Mice in another group were sham-infected control. Three mice from each group were randomly chosen to be sacrificed on days 3, 7, 14 and 28 after the infection. Viral titers in spleen tissues were determined using a standard plaque assay. The expression of IL-17A mRNA and MCMV mRNA in the splenocytes were measured by RT- PCR. The expression of IL-17A in spleen tissues was observed by immunohistochemica1 staining. The pathology of the infected mice was assessed by histological examination of H＆E stained spleen sections. Results Viral titers and MCMV mRNA in the spleen peaked on day 3, but quickly dimin- ished on day 7. Virus was no longer detectable in the spleen on day 14 after the infection. The expression of IL-17A mRNA was significantly increased during the acute infection and reached the highest level on day 14, then decreased on day 28. It is significantly higher than that of the mock infection group. Immunohistochemistry assay also indicated that the expression of IL-17A in spleen tissue gradually increased to climax on day 14, then decreased on day 28. Accordingly, the pathological damages of spleen tissue in the infected mice deteriorated until day 14, then showed signs of recovery on day 28. The most severe pathological injury of spleen tissue and the highest expression of IL-17A appeared in the same period of time. Conclusion Our results showed a close correlation between IL-17A and the pathological damage in spleen. Thus, IL-17A may contribute to the spleen pathological damage during the acute disseminated MCMV infection.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2013年第3期188-192,共5页
Chinese Journal of Microbiology and Immunology
基金
教育部博士点基金（20090142110076）
国家自然科学基金（81271870）
作者简介
方峰，Email：ffang@tjh．tjmu．edu．cn，电话：027-83663579
