以CotB为分子载体表面展示鸡白痢沙门氏菌OmpC的重组枯草杆菌芽孢对小鼠免疫效果的研究  

Immune effect in mice administered orally with recombinant Bacillus subtilis spores expressing OmpC of Salmonella pullorum by using CotB as a fusion partner

作　　者：刘明刚[1] 戴茜茜[1] 徐毓琴 张飞燕[1] 唐慧琴[1] 潘康成[1,2] 

机构地区：[1]四川农业大学动物医学院/动物微生态研究中心,四川成都611130 [2]动物疫病与人类健康四川省重点实验室,四川成都611130

出　　处：《中国预防兽医学报》2015年第12期966-969,977,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金："教育部长江学者和创新团队发展计划"创新团队资助项目(IRT0848)

摘　　要：为研究小鼠口服以CotB为分子载体表面展示鸡白痢沙门氏菌外膜蛋白OmpC的重组枯草杆菌芽孢后诱导产生的特异性抗体水平与交叉保护作用,本研究将120只成年雌性小鼠随机分为5组,A组与B组在第0、15 d分别灌服重组枯草杆菌SE1与野生型枯草杆菌168芽孢(1.0×10^(10)cfu芽孢/次/只),C组与D组则分别拌料饲喂重组枯草杆菌SE1与野生型枯草杆菌168芽孢(1.0×10~6cfu芽孢/g饲料),E组仅饲喂基础日粮。饲喂至22d,每组随机选取4只小鼠采集血清和小肠内容物,采用ELISA检测各组小鼠OmpC特异性抗体水平;同时,每组小鼠随机选取16只并分为两个小组,进行鼠伤寒沙门氏菌交叉保护试验。结果显示,口服重组芽孢可诱导产生特异性血清IgG与肠粘膜SIgA抗体,与对照组相比差异极显著(p<0.01),拌料饲喂方式所诱导的抗体水平均显著高于灌服方式(p<0.05);重组芽孢可为小鼠抗2×LD_(50)和10×LD_(50)攻毒剂量的鼠伤寒沙门氏菌感染提供100%和50%的保护作用。结果表明,重组芽孢经口服免疫可诱导机体产生特异性血清IgG与肠粘膜SIgA抗体,并可赋予交叉保护作用抗鼠伤寒沙门氏菌感染。To assess specific antibodies levels and cross-protective activities in mice administered orally with recombinant Bacillus subtilis spores expressing the OmpC antigen fused to the spore coat protein CotB, a total of 120 mice were randomly divided into 5 groups： group A and B were administrated orally with spores of recombinant B.subtilis SE1 or isogenic wild-type B.subtilis 168 by gavage （1.0×10-10 cfu spores/time/mouse） on day 0 and 15; group C and D were dosed orally by feeding the diet mixed with spores of the recombinant or wild-type strains （1.0×106 cfu spores/g）; a naive, untreated control group was included （group E）. The levels of the OmpC-specific antibodies in the serum and small intestinal content samples collected on 22 day were determined by ELISA. In addition, 16 mice of each group were randomly chosen and divided into two subgroups, and cross protection against Salmonella typhimurium were also explored. The results showed that antibody levels of both specific serum IgGand intestinal mucosa SIgA from mice dosed with recombinant B.subtilis SE1 spores were significantly higher than that from mice in control groups （p〈0.01）. Moreover, mice dosed by feeding the diet mixed with recombinant B.subtilis SE1 spores developed higher （p〈0.05） specific antibodies levels than mice dosed by gavage with spores of the same strain. The challenge of S.typhi- murium showed that mice dosed orally with recombinant B.subtilis SE1 spores provided 100% and 50% protection against a challenge dose of 2×LD50 and 10×LD50, respectively. The findings indicated that the recombinant B.subtilis SE1 spores was able to induce both specific serum IgG and intestinal mucosa SIgA and confer cross-protective activities against S. typhimurium infection following oral administration.

关 键 词：鸡白痢沙门氏菌 外膜蛋白OmpC 枯草芽孢杆菌 芽孢表面展示 免疫效果 

分 类 号：S852.61[农业科学—基础兽医学]