李乐,孙军峰,陈林娜,师乾凯,邸涛,刘添仪,赵冉,张春玮,韩宗玺,刘胜旺

• 1:中国农业科学院哈尔滨兽医研究所禽呼吸道传染病创新团队兽医生物技术国家重点实验室

摘要(Abstract)：

为探索新城疫病毒(NDV)F基因及其蛋白诱导细胞免疫的能力,本研究对基因Ⅶ型NDV CK/CH/LHLJ/1/06株F基因进行了密码子优化和修饰,将其克隆至真核表达载体,获得了重组质粒pCAF。将质粒pCAF转染至悬浮培养的细胞中,上清中表达的蛋白经纯化获得了重组蛋白proF。以构建的质粒pCAF及其重组蛋白proF作为DNA和亚单位疫苗,采用pCAF单独免疫、proF单独免疫、pCAF和proF联合免疫的方式分别免疫SPF鸡,免疫后分离外周血淋巴细胞,通过淋巴细胞增殖试验和细胞因子检测试验评估pCAF和proF诱导的细胞免疫反应。结果显示,proF可刺激分离自pCAF单独免疫组或proF单独免疫组的外周血淋巴细胞增殖和细胞因子IL-4和IFN-γ的分泌,并且proF刺激后,pCAF和proF联合免疫组的外周血淋巴细胞的增殖水平、细胞因子IL-4和IFN-γ的分泌水平显著高于pCAF单独免疫组或proF单独免疫组。上述结果表明,pCAF和proF均可诱导细胞免疫应答,且与pCAF或proF单独免疫相比,pCAF和proF联合免疫组诱导的免疫效果更好。本研究结果为研制新型NDV疫苗提供了思路和理论基础。

关键词(KeyWords)： 基因Ⅶ型新城疫病毒;F蛋白;淋巴细胞增殖;细胞因子

Abstract:

Keywords:

基金项目(Foundation): 国家重点研发计划资助项目(2017YFD0500805);国家重点研发计划资助项目(2018YFD0501302);; 国家自然科学基金资助项目(31872504)

作者(Author): 李乐,孙军峰,陈林娜,师乾凯,邸涛,刘添仪,赵冉,张春玮,韩宗玺,刘胜旺

DOI: 10.16303/j.cnki.1005-4545.2021.10.01

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