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首页> 外文期刊>Infection and immunity >Protection against Mycobacterium aviumby DNA Vaccines Expressing Mycobacterial Antigens as Fusion Proteins with Green Fluorescent Protein
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Protection against Mycobacterium aviumby DNA Vaccines Expressing Mycobacterial Antigens as Fusion Proteins with Green Fluorescent Protein

机译：表达绿色分枝杆菌抗原作为融合蛋白与绿色荧光蛋白的鸟分枝杆菌DNA疫苗的保护

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Mycobacterium avium causes disseminated disease in humans with AIDS, paratuberculosis in ruminants, lymphadenopathy in swine, and tuberculosis in birds. We constructed DNA vaccines expressing mycobacterial antigens as fusion proteins with enhanced green fluorescent protein (EGFP). Plasmids p65K-EGFP, p85A-EGFP, and p85B-EGFP expressed the M. avium 65-kDa antigen, theMycobacterium bovis BCG 85A antigen, and the M. avium 85B antigen, respectively, as EGFP fusion proteins. We visualized protein expression directly in cultured murine fibroblasts and intact muscle. p65K-EGFP expressed fusion protein in a diffuse cytoplasmic pattern, and p85A-EGFP and p85B-EGFP produced a speckled pattern. We vaccinated C57BL/6 mice with three doses of plasmid DNA and then challenged them intraperitoneally with M. avium. Negative controls received saline, and positive controls received one dose of BCG vaccine. Mice in all groups developed disseminated infection with a high burden of organisms. Compared to negative controls, mice vaccinated with p85A-EGFP had an eightfold reduction in spleen M. avium CFU at 4 weeks after infection and a fourfold reduction at 8 weeks, reductions similar to those generated by BCG vaccine. Mice vaccinated with p65K-EGFP had a fourfold CFU reduction at 4 weeks and no effect at 8 weeks. This is the first report of DNA vaccines expressing foreign antigens as fusion proteins with EGFP and the first report of successful DNA vaccination againstM. avium.
机译：鸟分枝杆菌在艾滋病患者，反刍动物的副结核病，猪的淋巴结病和鸟类的结核病中引起传播性疾病。我们构建了表达分枝杆菌抗原的DNA疫苗，将其与增强的绿色荧光蛋白（EGFP）融合在一起。质粒p65K-EGFP，p85A-EGFP和p85B-EGFP表达 M。禽类 65 kDa抗原，牛分枝杆菌BCG 85A抗原和 M。 avium 85B抗原分别作为EGFP融合蛋白。我们直接在培养的鼠成纤维细胞和完整的肌肉中可视化了蛋白质表达。 p65K-EGFP以弥散的细胞质模式表达融合蛋白，p85A-EGFP和p85B-EGFP产生斑点模式。我们给C57BL / 6小鼠接种了三剂质粒DNA，然后用 M腹膜内攻击它们。 avium 。阴性对照接受盐水，阳性对照接受一剂卡介苗疫苗。所有组中的小鼠都发生了传播性感染，并伴有大量生物。与阴性对照相比，接种p85A-EGFP的小鼠脾脏M降低了八倍。感染后第4周的鸟粪CFU和第8周的4倍减少，与BCG疫苗产生的减少相似。接种p65K-EGFP的小鼠在4周时CFU降低了4倍，而在8周时没有效果。这是表达外源抗原与EGFP融合蛋白的DNA疫苗的首次报道，也是成功针对em进行DNA疫苗接种的首次报道。 avium 。

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