表达猪传染性胃肠炎病毒S蛋白的重组乳酸菌口服免疫小鼠特异性免疫应答分析

将表达猪传染性胃肠炎病毒S蛋白的重组乳酸乳球菌pNZ8112-Sa/NZ9000经口服免疫BALB/c小鼠,在免疫后的不同时间收集免疫鼠粪便样品,断尾采血收集血液样品并分离血清,用间接ELISA技术检测血清中的IgG抗体及血清和粪便中的IgA抗体的动态产生规律;在加强免疫后流式细胞仪检测分析外周血T细胞的变化情况及无菌收集免疫鼠的肠黏液,经中和试验检测其抗体的中和能力。结果表明重组菌株口服免疫小鼠后血清中的IgG和IgA抗体产生能力由于非特异性干扰不能确定,外周血中T细胞百分数在实验组和对照组间变化不显著;粪便中的IgA抗体水平产生效果明显;黏液的抗体具有一定的中和能力,其效价检测结果为1:36。

Immune response of mice following oral inoculation of recombinant lactobacillus expressing porcine transmissible gastroenteritis virus spike glycoprotein

BALB/c mice were orally immunized with recombinant lactococcus lactis pNZ8112-Sa/NZ900 strain that expresses porcine transmissible gastroenteritis virus(TGEV)Spike(S)glycoprotein.Feces and serum samples were collected on different days after immunization,IgG,IgA in serum and slgA in the feces were examined by indirect ELISA.Changes of T cells proportion in peripheral blood lymphocytes in mice were analyzed by the flow cytometry.The neutralization ability of the aseptically collected intestinal mucin from mice was valuated after the third inoculation.The production of the IgG and IgA antibody from the orally immunized mice could not be determined due to nonspecific interference.The proportion of CD4~ ,CD8~ T cells in peripheral blood lymphocytes in mice showed no difference between the experimental and control groups.Neutralization test showed a neutralizing activity of 1:36 from the intestinal lavage.