以小鼠为模型评价重组伪狂犬病病毒rPRV-HA的免疫效力  

Efficacy Evaluation of a Recombinant Pseudorabies Virus Expressing the Hemagglutinin Gene from H3N2 Subtype Swine Influenza Virus as Vaccine in a Mouse Model

作　　者：郑宝亮[1] 周国辉[1] 田志军[1] 仇华吉[1] 杨焕良[1] 尹训南[1] 胡守萍[1] 童光志[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室

出　　处：《畜牧兽医学报》2006年第4期361-367,共7页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家科技攻关项目(2004BA519A21);国家"973"项目(2005CB523200)

摘　　要：以小鼠为动物模型,对此前构建的表达H3N2亚型猪流感病毒(SIV)血凝素(HA)基因的重组伪狂犬病病毒(rPRV-HA)进行了免疫效力评价。按每只105.0TCID50rPRV-HA的剂量通过滴鼻接种8周龄雌性BALB/c小鼠(n=60),同时设Bartha-K61免疫对照组(n=60)、非免疫攻毒对照组(n=20)和非免疫不攻毒对照组(n=10)。于免疫后不同时间分别从rPRV-HA免疫组和Bartha-K61免疫对照组随机剖杀一定数量的小鼠,其余小鼠于免疫后第28天用105.0TCID50同亚型SIV毒株A/Swine/Heilongjiang/74/2000(H3N2)进行强毒攻击。攻毒后第4、7、14天分别剖杀小鼠,进行间接免疫荧光、病毒分离、血清学和病理组织学检测。结果表明,重组病毒主要分布于肺脏;免疫后14 d起,从rPRV-HA免疫组及Bartha-K61免疫对照组均可检测到针对PRV的荧光抗体;从rPRV-HA免疫组可以检测到针对SIV的荧光抗体和血凝抑制抗体,而各对照组均呈阴性。攻毒后从rPRV-HA免疫组小鼠未分离到攻击病毒,血凝抑制抗体显著升高,病理变化显著轻于对照组,表明rPRV-HA免疫小鼠可以抵抗同亚型SIV的攻击,可以作为rPRV-HA免疫效力评价模型。A recombinant pseudorabies virus （PRV） （rPRV-HA） expressing the hemagglutinin （HA） gene from H3N2 subtype swine influenza virus （SIV） was evaluated in a mouse model. A total of 150 8-week-old female BALB/c mice were used in this study. The mice were each inocula ted intranasally with 10^5.0 TCID50 of rPRV-HA （rPRV-HA group, n=60） or attenuated vaccine Bartha-K61 （Bartha-K61 group, n=60）, and another two unimmunized groups served as challenged group （n=20） or unchallenged group （n= 10）. Some mice in rPRV-HA and Bartha K61 groups were sacrificed for antigen detection and virus isolation at different days post-immunization （DPI）, and the others were each challenged with 10^5.0 TCID50 of the same subtype SIV 28 DPI.Recombinant virus could be detected in the lung of rPRV-HA-immunized mice. Antibody responses to PRV were detected by indirect immunofluorescence assay, but not by sero-neutralization test, in both rPRV HA and Bartha-K61 groups. SIV-specific antibodies were detected by hemagglutination inhibition test only in rPRV-HA group 14 DPI. The rPRV-HA-immunized mice were protected from homologous SIV challenge, as indicated by limiting virus replication and pathological changes of the organs, and boosted antibody responses to SIV post-challenge.

关 键 词：伪狂犬病病毒 H3N2亚型猪流感病毒 重组病毒 小鼠模型 免疫效力 

分 类 号：S858.285.3[农业科学—临床兽医学] S852.52[农业科学—兽医学]