【摘要】 利用分子克隆、定点突变以及重组表达技术获得猪肺炎支原体的主要免疫原蛋白p36、p46、p65和p97R1-Nrdf。设立以重组蛋白p36、p46和p65为试验Ⅰ组,以p36、p46、p65和p97R1-Nrdf为试验Ⅱ组,猪支原体疫苗安百克(M+PAC)为试验Ⅲ组,PBS+佐剂为空白对照组来免疫BALB/c小鼠。检测小鼠血清、肺脏以及经蛋白刺激脾淋巴细胞的猪肺炎支原体抗体、IFN-γ和IL-4。结果显示,Ⅱ组的猪肺炎支原体抗体水平极显著高于Ⅰ、Ⅲ组(P<0.01);Ⅱ组IFN-γ水平极显著高于Ⅲ组,而Ⅱ组与Ⅰ组、Ⅰ组与Ⅲ组的IFN-γ水平无显著差异(P<0.05);但Ⅰ、Ⅱ、Ⅲ组的IL-4水平差异不显著(P<0.05),但都极显著高于对照组(P<0.01);肺的检测结果中猪肺炎支原体抗体、IFN-γ和IL-4水平呈现一致性,都为Ⅱ组>Ⅰ组>Ⅲ组>对照组;而经刺激脾淋巴细胞检测结果为Ⅱ组的猪肺炎支原体抗体和IFN-γ水平最高,而IL-4水平为Ⅲ组最高。由此可见,Ⅱ组通过激活体液免疫和细胞免疫途径,产生的猪支原体肺炎抗体水平最高;Ⅰ组亦能通过这2种免疫途径达到与Ⅲ组同样的免疫效果。更多还原

【Abstract】 To construct a multi-subunit vaccine that could provide favorable immunologic efficacy,which provides basic data for the research of developing novel vaccines of Mycoplasmal Pneumonia of swine.The main immune original proteins of Mycoplasma hyopneumoniae p36,p46,p65 and p97R1-Nrdf were recombinantly expressed by using molecular cloning,directed mutations and restructuring expression techniques.Using the protein of p36,p46 and p65 as group Ⅰ,p36,p46,p65 and p97R1-Nrdf as group Ⅱ,using the Mycoplasma hyopneumoniae bacterin(M+PAC) as group Ⅲ,PBS added with adjuvants as the control group to immunize BALB/c mice.Detect serum,lungs and the splenic lymphocytes which is stimulated by protein with Mycoplasma pneumoniae antibody,IFN-gamma and IL-4.The results suggested that group Ⅱ induced significantly higher antibody than group Ⅰ and group Ⅲ(P<0.01) and also induced significantly higher IFN-γ production than group Ⅲ(P<0.05),however,there were no significant differences between group Ⅱ and group Ⅰ or group Ⅰ and group Ⅲ(P<0.05).IL-4 production was similar among the three group(P<0.05),but was obviously higher than the control group(P<0.01).The antibody level of MP antibody,IFN-γ and IL-4,in lung was group Ⅱ,group Ⅰ,group Ⅲ and the control group in proper order.The splenic lymphocytes results showed that the highest1 MP antibody and IFN-γ level were induced by group Ⅱ,while the highest IL-4 production was induced by group Ⅲ.It shows that the recombinant subunit vaccine containing p36,p46,p65 and p97R1-Nrdf provided the highest antibody level by simultaneously stimulating humoral immunity and cell immunity.Group Ⅰ provided almost the same significantly immunologic efficacy as group Ⅲ in the same way.更多还原