最新录用：H3N2亚型犬流感病毒实验感染模型的建立

doi:10.3864/j.issn.0578-1752.210651R

中国农业科学

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最新录用：H3N2亚型犬流感病毒实验感染模型的建立

黄程¹，杨龙峰²，孙鹏³，程慧敏¹，杨志远¹，林健¹，祝洪伟³，刘立新¹，孙厚民³，李加凤³，赵际成¹，段会娟¹，潘洁¹，刘月焕¹*

¹北京市农林科学院畜牧兽医研究所，北京 100097; ²北京市延庆区动物疫病预防控制中心，北京 102100; ³青岛易邦生物工程有限公司，山东青岛 266113

发布日期:2022-10-09

Establishment of a Canine Experimental Infection Model with a H3N2 Subtype Canine Influenza Virus

HUANG Cheng¹, YANG LongFeng², SUN Peng³, CHENG HuiMin¹, YANG ZhiYuan¹, LIN Jian¹, ZHU HongWe³i, LIU LiXin¹, SUN HouMin³, LI JiaFeng³, ZHAO JiCheng¹, DUAN HuiJuan¹, PAN Jie¹, LIU YueHuan¹* #br#

¹Institute of Animal Husbandry and Veterinary Medicine, Beijing Municipal Academy of Agricultural and Forestry Sciences，Beijing，100097; ²Yanqing District Animal Disease Prevention and control center of Beijing Municipality, Beijing, 102100; ³YEBIO Bioengineering Co., Ltd of Qingdao, Shandong, 266113

Online:2022-10-09

摘要/Abstract

摘要： 【目的】建立H3N2亚型犬流感病毒（canine influenza virus, CIV）实验感染模型，了解犬流感的发病特征，为疫苗效力评价奠定基础。【方法】6—13月龄CIV血凝抑制（haemagglutination inhibition, HI）抗体阴性（HI<1:10）比格犬26只，其中3只鼻腔喷雾PBS作为阴性对照，另23只分5组（10、10³、10⁵、10⁶、10⁷EID₅₀/只），分别为3、5、5、5、5只/组，每只犬各鼻腔喷雾H3N2亚型CIV（A/canine/China/Huabei-170607/2017(H3N2)，简称HB株）病毒液1mL。观察临床症状、肺脏病变和肺脏病理组织学变化，计算肺实变率，检测病毒和HI抗体效价。【结果】10 EID₅₀剂量感染组，3只犬均未出现明显临床症状，肺脏均无明显眼观病变，肺脏实变率0%，无病理组织学变化，病毒检测均为阴性，HI抗体效价几何平均值（geometric Mean Titer, GMT）为1:15.9；10³EID₅₀剂量感染组，5只犬中有4只喘息、流鼻汁和咳嗽，1只犬未表现出临床症状，2只犬肺脏出现轻微实变，实变率平均为1.4%，4只犬病毒分离阳性，HI抗体效价GMT为1:320；10⁵EID₅₀剂量感染组，5只犬在攻毒后5d开始出现流鼻汁和咳嗽等临床症状，肺脏均有实变，实变率为4.2%，肺泡间隔增宽，病毒分离均为阳性，HI抗体效价GMT为1:2940.7；10⁶EID₅₀剂量感染组，5只犬在攻毒后4天体温升高、流鼻汁、咳嗽，临床症状出现较10⁵EID₅₀剂量感染组早1d，肺脏实变程度增加，实变率为17.9%，肺泡间隔增宽，病毒分离均为阳性，HI抗体效价GMT为1:2228.7；10⁷EID₅₀剂量感染组，5只犬在攻毒后3d表现出体温升高、流鼻汁、咳嗽等严重的临床症状，症状出现较10⁶EID₅₀剂量感染组早1d，肺脏严重实变，实变率为29.0%，肺泡间隔增宽，病毒分离均为阳性，HI抗体效价GMT为1:2940.7；对照犬均未出现明显临床症状，肺脏均无明显眼观病变，无病理组织学变化，病毒检测均为阴性，HI抗体效价均＜1:10。【结论】10⁶EID₅₀剂量病毒是能引起犬明显发病的最小病毒接种剂量，建立起H3N2亚型CIV实验感染模型。

关键词: 比格犬, 犬流感病毒, H3N2亚型, 肺脏, 感染模型

Abstract: 【Objective】 In this research, an experimental animal infection model of canine influenza virus (CIV, H3N2 subtype) was established to better understand the pathogenesis of canine influenza and to lay the foundation for vaccine efficacy evaluation. 【Methods】 26 beagles aged 6-13 months with negative CIV Haemagglutination Inhibition (HI) antibody (HI < 1:10) were selected. Three were challenged by 1mL nasal spray of PBS. And twenty-three challenged by 1mL nasal spray of H3N2 CIV (A/canine/China/Huabei-170607/2017(H3N2), HB strain for short) with 5 groups (10、10³、10⁵、10⁶、10⁷ 50% EID₅₀) as 3, 5, 5, 5 and 5 beagles each group respectively_. Clinical symptoms, lung lesions, histopathological changes of lung, calculating the proportion of consolidation mass, HI antibody titer and virus shedding were examined 14 days after virus challenge together with three control beagles. 【Results】 3 beagles inoculated with a dose of 10 EID₅₀ H3N2 CIV did not show any clinical symptoms , gross lesions in lungs and histopathological changes. The consolidation rate was 0%. Virus shedding was not detected. The geometric mean titer (GMT) of HI antibodies was 1:15.9. However, 4/5 of beagles inoculated with a dose of 10³ EID₅₀ H3N2 CIV showed clinical symptoms and virus shedding such as puffing, runny nose and cough. 2/5 of beagles showed light lung consolidation, whose rate was 1.4%. The GMT of HI antibodies was 1:320. All beagles (5/5) infected with a dose of 10⁵ EID₅₀ H3N2 CIV showed clinical symptoms at day 5 after challenge such as runny nose and cough, virus shedding, lung consolidation and widened alveolar septum. The consolidation rate was 4.2%. The GMT of HI antibodies was 1:2940.7. 5/5 of beagles infected with a dose of 10⁶ EID₅₀ all showed severe clinical symptoms at day 4 after challenge such as cough and elevated body temperature, virus shedding, obvious pathological features in the lungs with GMT of HI antibodies were 1:2228.7. The clinical symptoms appeared earlier 1 day than that in 10⁵ EID₅₀dose infection group. And the degree of lung consolidation increased. The lung consolidation ratio was 17.9%. 5/5 of beagles infected with doses of 10⁷ EID₅₀ all showed severe clinical symptoms at day 3 after challenge which appeared earlier 1 day than that in 10⁶ EID₅₀dose infection group, virus shedding and widened alveolar septum in the lungs with GMT of HI antibodies were 1:2228.7. The consolidation rate of lung was 29.0%. Control beagles did not show any clinical symptoms, gross lesions and histopathological changes in lungs. And virus shedding was not detected. GMT of HI antibodies was ＜1:10. 【Conclusions】 The dose of 10⁶ EID⁵⁰ is the minimum virus inoculation dose that can cause obvious pathogenesis in beagles. An experimental animal infection model of CIV subtype H3N2 in beagles was established.

Key words: beagles, canine influenza virus, H3N2 subtype, lung, infection model

黄程, 杨龙峰, 孙鹏, 程慧敏, 杨志远, 林健, 祝洪伟, 刘立新, 孙厚民, 李加凤, 赵际成, 段会娟, 潘洁, 刘月焕. 最新录用：H3N2亚型犬流感病毒实验感染模型的建立[J]. 中国农业科学, 0, (): 1-.

HUANG Cheng, YANG LongFeng, SUN Peng, CHENG HuiMin, YANG ZhiYuan, LIN Jian, ZHU HongWei, LIU LiXin, SUN HouMin, LI JiaFeng, ZHAO JiCheng, DUAN HuiJuan, PAN Jie, LIU YueHuan. Establishment of a Canine Experimental Infection Model with a H3N2 Subtype Canine Influenza Virus[J]. Scientia Agricultura Sinica, 0, (): 1-.

[1]	黄程, 杨龙峰, 孙鹏, 程慧敏, 杨志远, 林健, 祝洪伟, 刘立新, 孙厚民, 李加凤, 赵际成, 段会娟, 潘洁, 刘月焕. H3N2亚型犬流感病毒实验感染模型的建立[J]. 中国农业科学, 2023, 56(13): 2620-2628.
[2]	阿依木古丽·阿不都热依木, 张晨, 蔡勇, 覃圣, 罗文学, 扎西英派. 藏绵羊肺脏结构特点及HIF-1α和AQP1的表达特性[J]. 中国农业科学, 2023, 56(11): 2202-2211.
[3]	陈秋生,冯霞. 牦牛肺脏的适应性结构[J]. 中国农业科学, 2006, 39(10): 2107-2113 .

最新录用：H3N2亚型犬流感病毒实验感染模型的建立

Establishment of a Canine Experimental Infection Model with a H3N2 Subtype Canine Influenza Virus

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