pcv2b1a pcv2b1b pcv2b1c安徽分离株免疫原性比较分析-comparative analysis of immunogenicity of pc v2 b1 a pc v2 b1b pc v2 b1 c anhui isolate.docxVIP

免疫组之间 IgG 水平和细胞因子含量也无显著差异(P＞0.05)。结果表明： 3 株 PCV2 安徽株均能诱导机体产生较高水平的体液免疫应答，但细 胞免疫应答受到抑制，且诱导体液和细胞免疫应答的能力存在一定差异，其中PCV2-BH0801 与 PCV2-XC0801 免疫应答差异较明显。3 株 PCV2 安徽株 Cap 蛋白均 具有良好的免疫原性，能诱导机体产生较高水平的体液免疫应答，但诱导体液和细胞 免疫应答能力无明显差异，PCV2-XC0801 与 PCV2-BH0801 和 PCV2-DY0801 相比 ORF2 氨基酸序列位点发生的改变未导致其编码 Cap 蛋白免疫原性的明显改变。因此， 同一基因亚型不同基因簇的 PCV2 安徽株免疫原性存在一定的差异，但 Cap 蛋白免疫 原性无明显差异，造成毒株间免疫原性存在差异的原因不只是与 ORF2 氨基酸序列改 变有关，还与其他因素有关。关键词：猪圆环病毒 2 型，IgG，T 细胞亚群，Th1 /Th2 细胞因子，Cap 蛋白AbstractPorcine circovirus type 2 is the major pathogen of post-weaning multisystemic wasting syndrome (PMWS ), which divided into PCV2a, PCV2b and PCV2c genetic subtypes. In recent years, the herd infection mainly is PCV2b genetic subtypes.And the Anhui region PCV2 isolates were PCV2b, divided into PCV2b-1A, PCV2b-1B and PCV2b-1C gene cluster, the overall amino acid sequence of PCV2b-1A and PCV2b-1B are consistent, but compared with PCV2b-1 and PCV2b-1B ,the amino acid sequence of PCV2b-1C have more obvious changes,and the changes of loci focused on two immune reaction zone of ORF2.To explore the immunogenicity differences of the different cluster but same genetic subtype of porcine circovirus type 2 Anhui strains . In this study, the following content were studied, for PCV2 vaccine research and PCV2 pathogenesis of prevention and control measures provide a theoretical basis.Choose the different cluster but same genetic subtype of porcine circovirus type 2 strains (PCV2-DY0801 is PCV2b-1A, PCV2-BH0801 is PCV2b-1B, PCV2-XC0801 isPCV2b-1C) infect mice .To detect the IgG antibodies levels, the number of T cell subsets and Th1/Th2 cytokine levels in peripheral blood of mice by ELISA and flow cytometry.By expressing the PCV2 ORF2 which encode Cap protein and its immunological activity are identified.Primers are designed according to the ORF2 gene sequence of PCV2 Anhui strain form GenBank, ORF2 gene from PCV2 are amplified by PCR and clone into a pET-32α expression vector, generating the pET-32α-ORF2 construction. It is transformed