摘要
目的应用B族链球菌荚膜多糖-破伤风类毒素结合物(GBS-CPS-TT)及B族链球菌荚膜多糖-白喉类毒素结合物(GBS-CPS-DT),经不同免疫途径免疫小鼠,分别测定小鼠血清中特异性IgG的抗体滴度,观察并比较两种不同载体的B族链球菌荚膜多糖-蛋白质结合物经黏膜途径诱导的全身特异性免疫反应状况,探索最佳黏膜免疫途径及免疫剂量。
方法选取体重14-16g健康清洁级NIH品系雌性小鼠300只,随机取240只分别以GBS-CPS-TT、GBS-CPS-DT免疫,分为皮下注射5μg组、滴鼻5μg组、滴鼻10μg组和灌胃10μg组共8组,每组各30只。各组均免疫3次,每次免疫间隔时间2周,免疫后1周时采血,分离血清,低温保存。采用间接酶联免疫实验(ELISA)检测小鼠血清中特异性IgG的抗体滴度。
结果GBS-CPS-TT、GBS-CPS-DT结合物滴鼻免疫、灌胃免疫和皮下注射均可以诱导产生特异性血清IgG抗体。滴鼻10μg组诱导产生的IgG抗体滴度明显高于滴鼻5μg组(P<0.01)；当免疫剂量相同时,滴鼻免疫诱导产生的IgG抗体滴度显著低于皮下注射产生的IgG抗体滴度(P<0.01)。作为载体蛋白TT与DT比较无差异(P>0.05)。对照组未检出特异性IgG抗体。
结论GBS-CPS-TT、GBS-CPS-TT结合物经滴鼻免疫小鼠后可产生明显的系统性免疫应答,滴鼻10μg组的免疫效果优于5μg组,抗体滴度有显著性差异意义,表明较大的免疫剂量达到的免疫效果可能更优；黏膜免疫后第二、三针诱导应答产生的抗体滴度具有明显的递增增强效应,表明结合物有免疫记忆及加强免疫应答效应；GBS多糖蛋白质结合物的黏膜免疫机制还有待进一步研究。
Objectives To observe the effect to the immunogenicity of different mucosal administration routes of GBS-CPS-TT、GBS-CPS-DT conjugate vaccines.
Methods NIH mice were divided into 8 groups,30 mice per group. Mice were immunized respectively with GBS-CPS-TT、GBS-CPS-DT conjugate vaccines 3 times with an interval of 2 weeks by intranasal or intragastric or hypodermic injection route. The amounts of IgG against GBS were measured by ELISA.
Results The levels of IgG in serum were significantly increased after intranasal immunization. The levels of IgG of the group of 10μg intranasal immunization was significantly higher than the group of 5μg intranasal immunization(P<0.01). It was also shown that immunological memory and boosted response could be elicited by GBS-TT or GBS-DT conjugate vaccines. TT or DT shows no difference to be the carrier to make GBS-protein conjugate vaccines (P>0.05).The levels of IgG of the group of the control group can not be detected.
Conclusions GBS-CPS-TT、GBS-CPS-DT conjugate vaccines can induce system immunity reaction by intranasal immunization, the higher immunizing dose may achieve the better immunizing results and this method of immunization appears to be a safe and effective immunization route. The effect of immune responses induced by mucosal immunization needs more study.
引文
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