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目的观察感染旋毛虫小鼠IgE水平的动态变化及其抗体依赖细胞介导的细胞毒效应(antibody-dependent cell cytotoxicity,ADCC),并通过IL-4、IL-5的表达测定,探讨IgE在旋毛虫感染小鼠急性期免疫反应中的作用。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,同时用间接ELISA法测定特异性IgE水平,采用体外培养法做IgE介导嗜酸性粒细胞对旋毛虫肌幼虫的体外杀伤实验,按分组设计在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(Eos)、感染鼠血清及感染鼠的旋毛虫肌幼虫与含有10%灭活小牛血清的RPMI-1640培养液,共培养2d。显微镜下,分别在12、24、48h观察小鼠Eos对旋毛虫肌幼虫的影响,另外,用RT-PCR方法观察感染旋毛虫小鼠脾脏中IL-4、IL-5的mRNA含量,观察这两种细胞因子的表达情况。结果实验组较对照组总IgE水平增高,在14、21d达到高峰(P<0.01),第3周后逐渐下降,特异性IgE在14和21d组与对照组比较差异有统计学意义(P<0.01),阳性率达88.9%。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后,Eos的杀伤作用明显减弱,与单纯Eos的情况无差别。感染组小鼠脾脏中IL-4和IL-5mRNA的含量均明显高于对照组。结论血清中IgE抗体参与了旋毛虫急性期的免疫,主要作用为快速排出肠道内的旋毛虫脱囊幼虫,Eos对旋毛虫肌幼虫杀伤的过程需要IgE的参与,且IgE是Eos发挥ADCC效应所依赖的重要免疫球蛋白,感染旋毛虫小鼠的IL-4和IL-5表达量明显升高,故说明IgE是小鼠旋毛虫感染急性期重要的抗体。
Objective To investigate the dynamic changes of IgE levels in mice infected with Trichinella spiralis and its antibody-dependent cell cytotoxicity (ADCC). To investigate the expression of IL-4 and IL-5, The role of caterpillar in immune response in acute phase of mice. Methods The total IgE levels in peripheral blood of mice infected with Trichinella were detected by double antibody sandwich ELISA. The levels of IgE in peripheral blood of mice infected with Trichinella were assayed by indirect ELISA. The in vitro culture method was used IgE-mediated eosinophils in vitro killing of Trichinella spiralis larvae experiment, according to the design of a group of 96-well cell culture plate plate wells infected with Trichinella infection in mice peripheral blood eosinophils (Eos), infected mice Serum and infected mice Trichinella muscle larvae and RPMI-1640 medium containing 10% inactivated calf serum were co-cultured for 2 days. The effects of mouse Eos on the larvae of Trichinella spiralis were observed under microscope at 12, 24 and 48 hours respectively. The mRNA levels of IL-4 and IL-5 in spleen of mice infected with Trichinella were observed by RT-PCR. The expression of two cytokines. Results The level of total IgE in experimental group was higher than that in control group, reached its peak on 14 and 21 days (P <0.01), and gradually decreased on the 3rd week. The specific IgE was significantly different between the 14th and 21st groups and the control group (P < 0.01), the positive rate of 88.9%. 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 significantly enhanced with the involvement of immune serum. The killing effect of Eos after inactivation of IgE was significantly weaker than that of Eos alone No difference. The content of IL-4 and IL-5mRNA in the spleen of infected mice were significantly higher than that of the control group. Conclusions Serum IgE antibody is involved in the acute phase of Trichinella spiralis immunization. Its main function is to rapidly excrete the trichinella spiralis larvae in the intestine. IgE is involved in the killing of Trichinella spiralis larvae by Eos, and IgE plays an important role in ADCC Dependent on the important immunoglobulin, IL-4 and IL-5 expression in mice infected with Trichinella significantly increased, indicating that IgE is an important antibody to mouse Trichinella infection in acute phase.