目的观察细粒棘球绦虫幼虫原头蚴及囊液对体外培养小鼠脾细胞产生IL-22的影响。方法取Balb/c小鼠脾细胞,分别加入不同浓度原头蚴或囊液共培养48 h,检测上清液IL-22表达量及细胞IL-22 m RNA的相对表达量。同浓度原头蚴或囊液分别在0、12、24、36和48 h收集细胞,流式细胞术检测CD4+IL-22+T细胞比例变化。RPMI 1640培养基与脾细胞共培养设为对照组。结果与对照组相比,ELISA和q RT-PCR检测显示在原头蚴为1 000/ml和囊液蛋白质量浓度为2.05 mg/ml时脾细胞产生IL-22增加;流式细胞术检测原头蚴组及囊液组CD4+IL-22+T细胞比例逐渐增加。结论原头蚴及囊液均可促进小鼠脾细胞产生IL-22增加,提示IL-22可能参与宿主防御细粒棘球蚴感染。
To observe the effect of Echinococcus granulosus protoscolices and hydatid fluid on IL-22 expression in mouse splenocytes in vitro, splenocytes were prepared from Balb/c mice and were co-cultured with different concentrations of protoscolices or hydatid fluid for 48 h, then the level of IL-22 in co-cultured supernatants were measured using ELISA. The mRNA extracted from cells and the relative level of IL-22 expression was detected using qRT-PCR; the percentage of CD4+IL-22+T cells was detected at different stages (0, 12, 24, 36, 48 h after infection) by flow cytometry. Spleen cells were co-cuhured with RPMI 1640 to serve as a control. ELISA and qRT-PCR indicated that cells co-cultured with 2.05 mg/ml hydatid fluid proteins or 1 000/ml protoscoleces would secrete more IL-22, as compared with the control group. FCM analysis showed that the percentage of CD4+ IL-22 +T cells was increased obviously in hydatid fluid or protoscolices treated group after 12 h treatment. In conclusion, the secretion of IL-22 increases significantly in mouse splenocytes treated with hydatid fluid or protoscoleces in vitro, suggesting that IL-22 may paly an important role in host immune response after Echinococcus granulosus infection.