共表达新城疫病毒F基因、传染性法氏囊病病毒VP_0基因的重组鸡痘病毒的抗原性和免疫原性

Immunogenicity and antigenicity of Recombinant Fowlpox Viruses Coexpression of NDV F And IBDV VP_0 Gene

以鸡痘病毒 (FPV) 2 82 E4 株 TK基因为侧翼 ,将 2个相同的复合启动子 ATI· p7.5× 2 0以反向方式连接 ,分别调控新城疫病毒 (NDV) F基因和传染性法氏囊病病毒 (IBDV) VP0 基因 ,得到重组转移质粒 p VP0 - 2 ATI· p7.5× 2 0 -F。然后将该重组转移质粒用脂质体转染预先感染 FPV 2 82 E4 株的鸡胚成纤维细胞 (CEF) ,在经 Brd U (5 -溴 -脱氧尿嘧啶 )加压处理后的 CEF上传代 ,用间接免疫荧光试验、Western blot检测 ,有 2株鸡痘病毒能同时表达 NDV F蛋白和IBDV VP0 蛋白。用这 2株重组鸡痘病毒刺种商品 L eghorn雏鸡 ,于刺种前及刺种后 1、2、3周对抗 NDV特异性抗体水平和淋巴细胞转化能力检测 ,结果发现 ,重组鸡痘病毒能激发机体产生良好的免疫反应。结果表明 。

A transfering vector was constructed by inserted NDV F gene and IBDV VP 0 gene, under the control of two synthetic promoters ATI·p7.5×20 which were conjugated at opposite direction, within the thymidine kinase (TK) gene of FPV 282E 4 strain. The resulting vector , pVP 0 2ATI·p7.5×20 F, was coinfected CEF cells with FPV 282E 4 strain. the intermediate recombinants were passaged in CEF cells in the presence of 5′ bromo deoxyuridine(5′ BrdU), and were then screened for the expression of NDV F protein by indirect immunofluorescene assay and identified the expresssion of NDV F and IBDV VP 0 gene by western blot. Wing web inoculation of commercial leghorn chickens resulted in development of nonspecific cellular immunity and development of antibodies to NDV. The results suggested that the recombinant Fowlpox virus vFV282 could be candidate vaccine against virulent NDV, IBDV and FPV.