抗幽门螺杆菌不同免疫途径免疫奶羊的研究

目的:以幽门螺杆菌(Helicobacterpylori,H.polyri,Hp)全菌抗原免疫奶羊,观察羊奶中特异性抗体产生规律。方法:采用菌液浓度为60亿cfu/ml全菌抗原,通过滴鼻、多点皮下注射、妊娠期肌肉注射三种途径免疫3只产奶山羊。前两组都于0、14、21、28d接受四次免疫接种,妊娠组于分娩前后1个月每隔2w注射1次。收集血清、乳清样本,以酶联免疫吸附试验测定IgA、IgG抗体。结果:经滴鼻、多点皮下注射、妊娠期肌注三种途径免疫奶羊后产生了抗体。血清中IgA变化幅度不大,三组血清中IgG较免疫前都有很大升高。羊奶中IgA抗体较免疫前都有升高,抗体水平:滴鼻组>妊娠组>皮下组,羊奶中IgG抗体水平:妊娠组IgG>皮下组>滴鼻组。结论:滴鼻粘膜免疫、多点皮下注射、妊娠期肌肉注射三种途径都能激发奶羊机体的免疫应答,使羊奶中抗Hp-IgA、抗Hp-IgG抗体水平升高,血清中IgG抗体水平升高,尤其是滴鼻免疫能够诱导机体同时产生不同部位的粘膜免疫和系统免疫,且所需的抗原量远远少于其他两组,是一个敏感、安全、可行有效的免疫途径。

STUDY ON DIFFERENT IMMUNIZATION ROUTES TO PREPARE IMMUNE MILK AGAINST HELICOBACTER PYLORI IN GOATS

Objective: To observe the effect of three different routes on the induction of immunity to whole bacterial antigen of Vac A+ Helicobacter pylori strain (NCTC11637). Methods:Two lactating goats and one pregnant goat were immunized by different routes which were intranasal,subcutaneous and intra-muscular injection with whole bacterial antigen of H.pylori (6×109cfu/ml) cultivated by solid culture medium in respective immune procedure. Serum and milk samples were collected and assayed by indirect enzyme-linked immunosorbent assay (ELISA) which used H.pylori lysate obtained by sonication and ultracentrifugation as coated antigen. The levels of anti-Hp IgG and IgA in serum and milk were determined. The control milk and serum were obtained from the non-immunized goat and the immunized goats before being immunized. Result:Three immune routes all induced systemic immune response.The results of ELISA proved that specific IgG in serum increased while IgA didn't increase significantly,and that the specific IgA and IgG in milk all increased to a greater or lesser extent. Among three routes, intranasal inoculation could induce systemic and local immune response in different mucosal sites. Conclusion:Three immunization routes all can induce systemic immune response. Intranasal inoculation can induce mucosal immune response. and may be a safe and effective immunization route.