EAE小鼠体内γδT细胞的提取和鉴定

Study on extraction and identification of γδT-cell in EAE mice

目的试图从蛋白脂质蛋白(Proteolipid protein,PLP)免疫后的SJL/J小鼠肠上皮间(iIEL)、淋巴结(LNC)中提取γδT细胞,并探讨γδT细胞在实验性自身免疫性脑脊髓炎(Experimental autoimmune encephalomyelitis,EAE)发病机制中的作用。方法用多肽PLP178-191免疫小鼠,从小鼠iIEL、LNC中提取γδT细胞,利用免疫荧光标记和流式细胞分析技术进行检测。结果PLP免疫组和正常对照组比较,小鼠iIEL中CD3+细胞百分比在组间无统计学差异(P>0.05),而小鼠LNC中CD3+细胞百分比在组间存在显著性差异(P<0.01);小鼠iIELCD3+细胞中和LNCCD3+细胞中γδT细胞的百分比,组间均存在显著性差异(P<0.01)。结论利用PLP178-191免疫SJL/J小鼠,可以稳定地提取γδT细胞,γδT细胞的确参与了EAE发病过程。

Objective To extract γδT-cells from ilEL and LNC of SJL/J mice after immunized with PLP, and to explore its effect in EAE. Method PLP178-191 was used to immunize SJL/J mice, then γδT- cells were extractd from iIEL and LNC. Meanwhile, immunofluorescence marker and flow cytometry analysis were performed. Result Compared PLP group with control group, there were no marked different percentages of CD3^＋ cell in the ilEL （P〉0.05）. However, there were marked different percentages of CD3^＋ cell in the LNC （P〈0.05）, and marked different percentages of γδT-cells in the CD3^＋ cell both in the ilEL and LNC of SJL/J mice （P〈0.05）. Conclusion γδT-cell can be extracted from EAE model of SJL/J mice successfully, and γδT-cells indeed are correlated with EAE.