Recombinant turkey herpesvirus expressing H9 hemagglutinin providing protection against H9N2 avian influenza
Recombinant turkey herpesvirus expressing H9 hemagglutinin providing protection against H9N2 avian influenza
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DOI：
10.1016/j.virol.2019.01.004
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发表时间：
2019-03-01
期刊：
影响因子：
3.7
通讯作者：
Liu, Jinhua
中科院分区：
文献类型：
--
作者：
Liu, Litao;Wang, Tong;Liu, Jinhua
H9 N2禽流感病毒在鸡群中流行，给人类造成了巨大的经济损失和公共卫生威胁。本研究将火鸡疱疹病毒（HVT）克隆为具有传染性的细菌人工染色体（BAC）。通过半乳糖激酶（galK）筛选和规则间隔短回文重复序列/相关9（CRISPR/Cas9）基因编辑系统构建了含H9 N2病毒血凝素（HA）基因的重组HVT（rHVT-H9）。重组rHVT-H9与亲本HVT在鼠疫形态和病毒复制动力学上无差异。免疫印迹法和间接免疫荧光法（IFA）可检测rHVT-H9在体外和体内的H9蛋白表达。rHVT-H9免疫鸡可诱导机体产生较强的体液免疫和细胞免疫。攻毒试验中，各免疫组鸡口咽部和泄殖腔均未散毒，各免疫组鸡内脏均未检出H9 N2病毒。结果表明，rHVT-H9在鸡中提供针对H9 N2 AIV的有效保护。
H9N2 avian influenza viruses (AIVs) were prevailing in chickens, causing great economic losses and public health threats. In this study, turkey herpesviruses (HVT) was cloned as an infectious bacterial artificial chromosomes (BAC). Recombinant HVT (rHVT-H9) containing hemagglutinin (HA) gene from H9N2 virus were constructed via galactokinase (galK) selection and clustered regularly interspaced short palindromic repeats/associated 9 (CRISPR/Cas9) gene editing system. The recombinant rHVT-H9 showed no difference with parent HVT in plague morphology and virus replication kinetics. H9 protein expression of rHVT-H9 could be detected by western blot and indirect immunofluorescence assay (IFA) in vitro and in vivo. Immunization with rHVT-H9 could induce robust humoral and cellular immunity in chickens. In the challenge study, no chicken shed H9N2 virus from oropharynx and cloaca, and no H9N2 virus was found in viscera in vaccination groups. The result suggests that rHVT-H9 provides effective protection against H9N2 AIV in chickens.