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Positive effects of porcine IL-2 and IL-4 on virus-specific immune responses induced by the porcine reproductive and respiratory syndrome virus (PRRSV) ORF5 DNA vaccine in swine

机译:猪IL-2和IL-4对猪繁殖与呼吸综合征病毒(PRRSV)ORF5 DNA疫苗诱导的猪病毒特异性免疫应答的积极影响

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The purpose of this study was to investigate the effects of porcine interleukin (IL)-2 and IL-4 genes on enhancing the immunogenicity of a porcine reproductive and respiratory syndrome virus ORF5 DNA vaccine in piglets. Eukaryotic expression plasmids pcDNA-ORF5, pcDNA-IL-2, and pcDNA-IL-4 were constructed and then expressed in Marc-145 cells. The effects of these genes were detected using an indirect immunofluorescent assay and reverse transcription polymerase chain reaction (RT-PCR). Characteristic fluorescence was observed at different times after pcDNA-ORF5 was expressed in the Marc-145 cells, and PCR products corresponding to ORF5, IL-2, and IL-4 genes were detected at 48 h. Based on these data, healthy piglets were injected intramuscularly with different combinations of the purified plasmids: pcDNA-ORF5 alone, pcDNA-ORF5 + pcDNA-IL-2, pcDNA-ORF5 + pcDNA-IL-4, and pcDNA-ORF5 + pcDNA-IL-4 + pcDNA-IL-2. The ensuing humoral immune responses, percentages of CD4(+) and CD8(+) T lymphocytes, proliferation indices, and interferon-gamma expression were analyzed. Results revealed that the piglets co-immunized with pcDNA-ORF5 + pcDNA-IL-4 + pcDNA-IL-2 plasmids developed significantly higher antibody titers and neutralizing antibody levels, had significantly increased levels of specific T lymphocyte proliferation, elevated percentages of CD4(+) and CD8(+) T lymphocytes, and significantly higher IFN-gamma production than the other inoculated pigs (p < 0.05).
机译:这项研究的目的是调查猪白介素(IL)-2和IL-4基因对增强猪繁殖与呼吸综合征病毒ORF5 DNA疫苗在仔猪中的免疫原性的影响。构建真核表达质粒pcDNA-ORF5,pcDNA-IL-2和pcDNA-IL-4,然后在Marc-145细胞中表达。使用间接免疫荧光测定和逆转录聚合酶链反应(RT-PCR)检测了这些基因的作用。在Marc-145细胞中表达pcDNA-ORF5后的不同时间观察到特征性荧光,并在48小时检测到对应于ORF5,IL-2和IL-4基因的PCR产物。根据这些数据,向健康的仔猪肌肉内注射纯化质粒的不同组合:单独的pcDNA-ORF5,pcDNA-ORF5 + pcDNA-IL-2,pcDNA-ORF5 + pcDNA-IL-4和pcDNA-ORF5 + pcDNA- IL-4 + pcDNA-IL-2。随后的体液免疫反应,CD4(+)和CD8(+)T淋巴细胞的百分比,增殖指数和干扰素γ表达进行了分析。结果显示,用pcDNA-ORF5 + pcDNA-IL-4 + pcDNA-IL-2质粒共同免疫的仔猪产生了更高的抗体滴度和中和抗体水平,特异性T淋巴细胞增殖水平显着升高,CD4百分比升高( +和CD8(+)T淋巴细胞,并且比其他接种的猪明显更高的IFN-γ产生量(p <0.05)。

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