目的初步探讨细粒棘球蚴重组铁蛋白rEgferritin免疫小鼠产生的抗细粒棘球蚴感染的免疫机制。方法 rEg-ferritin免疫ICR小鼠3次、同时设立佐剂组和对照组,第12w用细粒棘球蚴原头蚴对3组小鼠进行攻击感染,5个月后剖杀小鼠,检查各组小鼠腹腔内包囊个数,根据公式计算保护力。分别于0w,12w,32w取血,制备血清,同时取脾,制备淋巴细胞悬液。通过ELISA检测血清中的IgG及其抗体亚型、IgE及MTT;通过流式细胞仪检测脾淋巴细胞中CD4+及CD8+亚群百分比。结果 rEgferritin组小鼠血清中的IgG,IgG2a,IgG2b持续增高,与佐剂组相比具有显著性差异(P〈0.01);3组IgG3无差异。rEgferritin组免疫小鼠可诱导小鼠产生85.6%的保护力抵御细粒棘球蚴原头蚴的再次感染,与对照组相比具有显著性差异(P〈0.01);rEgferritin组和佐剂组CD4+T细胞和CD8+T细胞均增加,rEgferritin组CD4+T细胞增加更为明显,与佐剂组相比有显著性差异(P〈0.01)。rEgferritin组脾细胞在受到rEgferritin抗原和天然抗原刺激后,细胞增殖明显。结论细粒棘球蚴重组抗原rEgferritin通过诱导宿主产生体液免疫级细胞免疫应答的方式抵御细粒棘球蚴感染的,说明rEgferritin是具有研究价值和潜力的包虫病候选疫苗分子。
In order to initially investigate the effects of rEgferritin on the immune response of mice infected with hydatid cysts of Echinococcus granulosus,ICR mice were immunized with rEgferritin by thrice and challenged by protoscoleces of E.granulosus at 12 week.Mice were killed at 20 weeks after infection,and blood of mice were obtain at 0 week,12 week and 32 week prepare serum,The splenocytes were isolated and the percentages of the splenic CD+4 and CD+ 8T lymphocytes were measured by FCM.In every groups splenocytes were cultivated with stimulation of rEgferritin native antigen,ConA and normal nutrient.The levels of the supernatant of cultures were determined by ELISA.The adjuvant group and blank group were set up for comparison.It was found that the OD values of IgG,IgG2a and IgG2b of immunized group were all higher than that of the control group(P0.01),and the OD value of IgG3 was no significant difference among the three groups.Mice vaccinated with rEgferritin and challenged intraperitoneally with E.granulosus protoscoleces revealed significant protective efficacy up to 85.6%.The percentage of the splenic CD+4 and CD+8T lymphocytes were all increased in the immunized group of mice,but CD+4T lymphocytes of immunized group were obviously higher than adjuvant group and control group(P0.01),splenic lymphocytes cell of immunized rEgferritin group of mice were growthed obviously by induced with rEgferritin protein and native antigen of Echinococcus g ranulosus.These data indicated that rEgferritin protein immunized could can induce humoral immunity of mice against the hydatid cyst of E.g ranulosus.The rEgferritin might be a valuable candidate vaccine for against hydatid disease.