小鼠感染肝片吸虫早期IL-4/IFN-!及M2巨噬细胞标记分子的变化  

Levels of IL-4and markers of M2macrophages at early stage of Fasciola hepatica infection in mice

作　　者：罗洪林[1] 张文韬[1] 郭智莉[1] 周雪梅[1] 周容琼[2] 周作勇[2] 

机构地区：[1]西南大学动物科技学院预防兽医学重庆市重点建设实验室,重庆北碚400716 [2]西南大学荣昌校区动物医学系,重庆荣昌402460

出　　处：《中国兽医学报》2013年第8期1253-1258,共6页Chinese Journal of Veterinary Science

基　　金：中央高校基本科研业务费专项资金资助项目(XDJK2009B001)

摘　　要：为弄清肝片吸虫感染早期的主要细胞免疫类型及选择性激活巨噬细胞(M2或AAMΦ)标记分子的变化,本试验采用肝片吸虫囊蚴为感染源,经口分别感染雌性BALB/c野生型小鼠,运用特异性PCR鉴定成功感染小鼠后用间接ELISA对腹腔中IL-4和IFN-!的水平进行测定,并对细胞因子IL-4、转录因子GATA3和M2的标记蛋白Relm"、Ym1分子的mRNA进行荧光定量PCR检测。结果显示,在感染后1,3,5,7周,从所获取肝脏组织中扩增的ITS2片段条带清晰,大小正确,测序后确定为肝片吸虫ITS2序列,表明肝片吸虫感染成功。对腹腔中IL-4和IFN-!的水平测定表明感染组IL-4在感染后1,3,5周比对照组的显著增加(P=0.013,P<0.01,P=0.02),但在第7周时无显著差异。IL-4的水平显著高于IFN-!(P=0.011),而在第7周两者间无显著差异;对腹腔中IL-4、GATA3、Relm"和Ym1mRNA水平的测定结果表明,感染后IL-4和GATA3mRNA水平在第3周达到最高峰;于感染后1,3,5,7周,IL-4和GATA3mRNA水平都分别显著高于对照组(IL4:P=0.01,P=0.012,P=0.023,P=0.014;GATA3:P=0.011,P<0.01,P=0.032,P=0.014),但IL-4和GATA3间无显著差异;Relm"和Ym1mRNA水平都分别显著高于对照组(Relm":P<0.01,P<0.01,P<0.01,P=0.011;Ym1:P<0.01,P<0.01,P<0.01,P=0.012),且二者间无显著差异,并随时间推移mRNA水平都呈下降趋势,于第7周达到最低水平。本研究成功利用肝片吸虫-小鼠模型研究蠕虫感染早期细胞免疫类型及M2标记分子的变化,发现在肝片吸虫感染小鼠早期主要引起Th2为主的细胞免疫。IL-4和M2巨噬细胞标记分子Relm"和Ym1在感染后1,3,5,7周都显著增加且具有相似的变化趋势,但Relm"和Ym1在第1周的高表达可能受诸多因素的影响还有待深入研究。To address the dominant type of cellular immune responses and levels of M2 macropha- ges markers at the early stage of Fasciola hepatica infection, the wild type female BALB/c mice were infected with F. hepatica metacercariae and the species-specific PCR was applied to identify the infection of F. hepatica at different time post infection. The indirect ELISA and real-time PCR were used to test the levels of IL-4/IFN-γ, and mRNA levels of IL-4,GATA3,Relm a and Yml in pleural cavity at week 1,3,5 and 7 post infection, respectively. The results showed at each time point,the ITS2 sequence were successfully identified and the IL-4 was dominant with comparison to IFN-γ, at week 1,3 and 5, but no difference at week 7. The mRNA levels of IL-4 and GATA3 were significantly higher than the controls with no difference between both during infection. More-over, both were significantly increased at week 3 compared to the rest time points,respectively. The mRNA levels of Relm α and Yml were significantly higher than the controls and their levels at week 1 and 3 were significantly ences were found between week 1, higher than those of week 3 and 5. In summary, the F. 7, respectively, although no differ- hepatica induces Th2 bias cellular immune responses at the early stage of infection. Levels of IL-4, Relm ct and Yml keep the same trend at different time courses. However,high expression of Relm ot and Yml at week 1 post in- fection may be affected by complicated factors which need further investigations.

关 键 词：肝片吸虫 M2巨噬细胞 TH2细胞因子 Relm" Ym1 

分 类 号：S852.735[农业科学—基础兽医学] R535[农业科学—兽医学]