柔嫩艾美耳球虫感染鸡的盲肠上皮间淋巴细胞表达白介素17的动态变化

以1×104个柔嫩艾美耳球虫(Eimeria tenella)孢子化卵囊感染10日龄AA肉鸡,于感染后第24、48及72小时分离鸡盲肠上皮间淋巴细胞(IELs)。提取鸡盲肠IELs总RNA,用实时荧光定量PCR方法检测IL-17的表达动态;然后用FITC标记的抗鸡CD4单抗和PE标记的抗小鼠IL-17单抗作为抗体对盲肠IELs进行荧光抗体染色,再进行流式细胞术分析,检测了表达IL-17的CD4+盲肠IELs在柔嫩艾美耳球虫感染后的动态变化。实时荧光定量PCR结果表明,鸡在感染柔嫩艾美耳球虫后能够显著上调IL-17mR-NA的表达水平,流式细胞分析结果表明,在球虫感染后表达IL-17的CD4+细胞数量显著上升。这些结果表明,IL-17参与了宿主抵抗球虫感染的反应。

Dynamic changes of IL-17 expression in the intraepithelial lymphocytes of chickens infected with Eimeria tenella

Cecal intraepithelial lymphocytes(IELs) of 10-day-old Arbor Acre(AA) chickens infected with 1×104 E.tenella oocysts were isolated at hour 24,48 and 72 post-infection.The total RNA of cecal IELs was prepared and used as templates of real-time RT-PCR for evaluating the dynamic profiles of IL-17 expression in chickens after E.tenella infection.FITC-labelled mAb specific for chicken CD4 and PE-labelled mAb specific to murine IL-17A secretory cells were used to probe the dynamic patterns of chicken IL-17-producing CD4+ IELs cells subsets during the early stage of E.tenella infection by flow cytometry.The results showed chicken IL-17 expression was highly up-regulated post E.tenella infection in both real-time RT-PCR and flow cytometric assays,which suggest that chicken IL-17 was involved in the host defense against E.tenella infection.