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目的通过对小鼠总IgE、特异性IgE及依赖IgE的嗜酸性粒细胞(Eos)介导的ADCC效应的动态观察,进一步探讨嗜酸性粒细胞在旋毛虫病免疫中的作用。方法采用双抗体夹心ELISA法,分别检测感染旋毛虫后7、14、21、28、35、60d小鼠外周血中总IgE水平,用间接ELISA法测定特异性IgE水平;分别在96孔细胞培养板的板孔中加入感染旋毛虫小鼠外周血提取的嗜酸性粒细胞。感染鼠血清及感染鼠的旋毛虫肌幼虫与含10%灭活小牛血清的RPMI-1640培养液,37℃条件下培养,分别在12、24、48h显微镜下观察小鼠Eos对旋毛虫肌幼虫的影响。结果实验组较对照组总IgE水平增高,在感染后14、21d达到高峰(P<0.01),之后呈逐渐下降;特异性IgE阳性率在感染后14和21d为88.9%,对照组阳性率为0,差异有统计学意义(P<0.01)。在体外培养的条件下,单纯Eos对旋毛虫肌幼虫无杀伤作用,在免疫血清参与下,Eos对幼虫的杀伤作用明显增强;灭活IgE和补体后,Eos的杀伤作用明显减弱,与单纯Eos组比较肌幼虫死亡率差异无统计学意义(P>0.05)。结论 Eos对旋毛虫肌幼虫杀伤的过程需要免疫血清的参与,免疫血清中介导上述效应的物质为IgE和补体。
OBJECTIVE: To further investigate the role of eosinophils in trichinosis immunization by dynamically observing ADCC effects induced by total IgE, specific IgE and IgE-dependent eosinophils in mice. Methods The total IgE levels in peripheral blood of mice infected with Trichinella were detected by double antibody sandwich ELISA at 7, 14, 21, 28, 35 and 60 days respectively. The specific IgE levels were measured by indirect ELISA. Plates of plate wells were infected with infected Trichinella peripheral blood of mice eosinophils. Serum and infected mouse Trichinella spiralis larvae and RPMI-1640 medium containing 10% inactivated calf serum were cultured at 37 ℃. The expression of Eos on Trichinella spiralis muscle was observed under microscope at 12, 24 and 48 hours respectively. Effects of larvae. Results The total IgE level in the experimental group was higher than that in the control group, reaching the peak at 14 and 21 days after infection (P <0.01), and then gradually decreased. The specific IgE positive rate was 88.9% at 14 and 21 days after infection, the positive rate in the control group was 0, the difference was statistically significant (P <0.01). Under the conditions of in vitro culture, Eos alone had no killing effect on Trichinella spiralis larvae. The killing effect of Eos on larvae was enhanced with the involvement of immune serum. The killing effect of Eos was significantly weakened after inactivation of IgE and complement, There was no significant difference in mortality between groups (P> 0.05). Conclusions The process of Eos killing of Trichinella spiralis muscle larvae requires the involvement of immune serum, and the substances that mediate the above effects in immune serum are IgE and complement.