携带pVAX1-GPV-P32重组减毒沙门菌诱导山羊免疫应答研究

Study on Immunity Response in Goat Induced by the Attenuated Salmonella typhimuriumCarrying pVAX1-GPV-P32

为探讨携带pVAX1-GPV-P32重组减毒沙门菌诱导山羊免疫应答效果,将60只健康黑山羊随机分为3组,即重组细菌组、弱毒疫苗组和空白对照组,重组细菌组灌服重组减毒沙门菌,弱毒疫苗组股内侧皮内注射山羊痘弱毒疫苗,空白对照组不作任何处理;于不同时间段采集山羊血液、呼吸道/消化道分泌液及组织样本进行检测。结果:(1)在免疫后7、15、30和45d的山羊消化道特异性SIgA含量中,重组细菌组明显高于弱毒疫苗组(P<0.05),45d时到达最高值(18.172±0.122)μg·L-1;(2)在诱导产生特异性P32抗体上,重组细菌组略低于弱毒疫苗组,但差异不明显(P>0.05);(3)在特异性淋巴细胞转化效率上,免疫15d前重组细菌组略低于弱毒疫苗组,但免疫30d后重组细菌组高于弱毒疫苗组,但差异均不显著(P>0.05);(4)在诱导产生特异性细胞因子方面,重组细菌组山羊血清IL-2含量低于弱毒疫苗组,差异极显著(P<0.01),血清IL-5含量高于弱毒疫苗组,差异极显著(P<0.01),血清TNF-β高于弱毒疫苗组,差异显著(P<0.05),而血清IFN-γ和TNF-α含量与弱毒疫苗组差异不显著(P>0.05)。上述结果表明,构建的携带pVAX1-GPV-P32重组减毒沙门菌能诱导山羊产生较好的免疫应答反应。

The aim of this study was to study the immune response effect of the recombinant at- tenuated Salmonella carrying pVAX1-GPV-P32 to goats. The 60 black goats were randomly di- vided into three groups as recombinant bacteria group, attenuated vaccine group and the control group, which the goats were fed by the recombinant attenuated Salmonella in recombinant bacte- ria group, and injected by the goat pox attenuated vaccine in the attenuated vaccine group, but without any treatment in the control group. Then the samples of the blood and respiratory/diges- tive tract secretion and tissue were collected respectively in the different periods after immuniza-tion and used for detection and analysis. The results were showed as follows. （1）The contents of the specific SIgA in recombinant bacteria group was significant higher than that in the attenuated vaccine group （P〈0.05） in 7, 15, 30 and 45 d post-immunization （PI）, and which reached the highest value （18. 172＋0. 122 μg L-1） in 45 d; （2） The contents of anti-GPV P32 antibody in the recombinant bacteria group were slightly lower than that in the attenuated vaccine group, but the difference between groups was not significant （P〈0.05）; （3）The transformation efficiency of the specific lymphocyte in the recombinant bacteria group was slightly lower than that in the attenuated vaccine group from 0 to15 d PI, but higher than that in the attenuated vaccine group in 30 d and 45 d PI, but the difference was not significant （P〈0.05）. （4）The IL-2 content of the goat serum in the recombinant bacteria group were less than that in the attenuated vaccine group and the difference was very significant （P〈0.01）, and the IL-5 levels were higher than that in the attenuated vaccine group and the difference was very significant （P〈0. 01） too, and the TNF-β levels were higher than that in the attenuated vaccine group and the difference was signifi- cant （P〈0.05）, and the content of IFN-γ and TNF-a were not significant with that in the atten- uated vaccine group （P〉0.05）. These results indicated that the recombinant attenuated Salmo- nella carrying the pVAX1-GPV-P32 made the goat have a better immune responses.