现用重组AIV(H5+H7)三价灭活疫苗对近期H5和H7亚型AIV分离株的免疫效力研究  

Protection efficacy of the recombinant AIV(H5+H7) trivalent inactivated vaccine against the current circulating H5 and H7 subtype AIV isolates

作　　者：刘艳晶 麻琦 潘舒心 和新文 王燕 邓国华[1] 李雁冰[1] 施建忠[1] 包红梅[1] 毛胜刚 胡井雷 路通 刘景利 郭兴福 田国彬[1] 曾显营[1] 陈化兰[1] LIU Yan-jing;MA Qi;PAN Shu-xin;HE Xin-wen;WANG Yan;DENG Guo-hua;LI Yan-bing;SHI Jian-zhong;BAO Hong-mei;MAO Sheng-gang;HU Jing-lei;LU Tong;LIU Jing-li;GUO Xing-fu;TIAN Guo-bin;ZENG Xian-ying;CHEN Hua-lan(National Avian Influenza Reference Laboratory/State Key Lab of Veterinary Biotechnology,National Poultry Laboratory Animal Resource Center/Harbin Veterinary Research Institute,CAAS,Harbin 150069,China;Harbin Weike Biotechnology Company Limited,Harbin 150069,China)

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/国家禽流感参考实验室/国家禽类实验动物资源库,黑龙江哈尔滨150069 [2]哈尔滨维科生物技术有限公司,黑龙江哈尔滨150069

出　　处：《中国预防兽医学报》2022年第5期514-520,共7页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家自然科学基金面上项目(31672593);现代农业产业技术体系资金资助(CARS-41)。

摘　　要：为评估现用重组禽流感病毒(AIV)(H5+H7)三价灭活疫苗(H5N1 Re-11株+Re-12株,H7N9 H7-Re3株)对我国近期H5和H7亚型AIV分离株的免疫预防效果,本研究对60只3周龄SPF鸡以肌肉注射途径接种该疫苗(0.3 mL/只),另外,60只鸡注射PBS作为阴性对照组;免疫3周后分组,每10只免疫鸡和10只对照鸡为一组,共6组,采集各组鸡血清并检测针对6株AIV分离株(2020年~2021年分离的4株H5N6、1株H5N8和1株H7N9亚型AIV)的相应HI抗体效价。同时用上述6株AIV分离株作为抗原差异代表株,分别以105EID50的剂量进行攻毒试验,攻毒后5 d采集各组所有存活鸡的喉头和泄殖腔拭子样品,死亡鸡样品随时采集,进行排毒检测。HI抗体检测结果显示,免疫后3周,5组免疫鸡血清中针对H5亚型Re-11株和Re-12株抗原的HI抗体效价在8log2~8.6log2,针对4株H5N6 AIV分离株(DK/HuN/S11553/2020、 DK/GD/SC001/2020、 DK/FJ/S1424/20和GS/GZ/S1569/21)和1株H5N8 AIV分离株(WS/SX/4-1/20)的平均抗体效价分别为5.0log2、4.6 log2、4.3log2、3.8 log2、3.7log2;针对H7N9 AIV H7-Re3株的HI抗体效价为7.8log2,而针对H7N9 AIV分离株CK/YN/SD024/21的平均抗体效价为4.0log2;对照组鸡血清中针对疫苗株和所有分离株的HI抗体均为阴性。表明上述分离株与现用疫苗株存在较大抗原性差异。攻毒后14 d的观察期内,所有免疫组鸡均健活,且攻击3株H5N6和1株H7N9 AIV分离株的免疫组鸡均不排毒,但攻击1株H5N6 AIV分离株(GS/GZ/S1569/21)和H5N8 AIV分离株的免疫组分别有2/10和3/10只鸡的喉头拭子样品中检测到低水平的病毒;所有对照组鸡在攻毒后4 d内全部发病、死亡,且死亡对照组鸡的喉头和泄殖腔拭子样品均呈AIV阳性。以上结果表明,以近期野鸟或家禽中分离的H5和H7亚型AIV抗原变异株攻击各组鸡后,现用的H5+H7 AIV三价灭活疫苗可对免疫鸡提供无临床发病的保护,并可有效阻止分离株在鸡体内的复制。本研究为H5和H7三价疫苗的�To evaluate the protection efficacy of the currently used recombinant avian influenza virus(AIV)(H5+H7) trivalent inactivated vaccine(H5N1 Re-11/Re-12 strains and H7N9 H7-Re3 strain) against current circulating H5 and H7 subtype AIV isolates, 60 3-week-old SPF chickens were inoculated by intramuscular injection with l dose(0.3mL) of the vaccine, and 60 chickens were inoculated with PBS as the negative control. At 3 weeks post immunization, the serum samples were collected from each bird for HI antibody detection, and all the birds were randomly divided into 6 groups with 10 immunized birds and 10 control birds. All the birds were challenged with a dose of 105.0ELD50of six representative antigenic variants recently isolated between 2020 and 2021in China, respectively, including four H5N6 strains, one H5N8 strain and one H7N9 strain. The oropharyngeal and cloacal swabs were collected individually from the survival birds at 5 days post challenge or dead birds at the time of death for virus shedding detection. At 3 weeks post vaccination, the HI antibody titers against H5 subtype Re-11 and Re-12 strains ranged from 8.0log2 to8.6log2, and the average antibody titers against 4 strains of H5N6 AIV(DK/HuN/S11553/2020, DK/GD/SC001/2020, DK/FJ/S1424/20 and GS/GZ/S1569/21) and 1 strain of H5N8 AIV(WS/SX/4-1/20) were 5.0log2, 4.6log2, 4.3log2, 3.8log2, and 3.7log2,respectively. The HI antibody titer against the H7N9 AIV H7-Re3 strain was 7.8log2, while the average antibody titer against the H7N9 isolate CK/YN/SD024/21 was 4.0log2. No HI antibodies were detected in the control group. The results indicated that all the isolates had considerable antigenic difference with the vaccine strains. During the observation period of 14 days post challenge, all the birds in the immunized group were survived without clinical signs, and the immunized birds challenged with 3 of H5N6 strains and one H7N9 strain were no virus shedding. However, low levels of virus were detected in oropharyngeal swabs from 2/10 and3/10 birds challenged with H5

关 键 词：禽流感 H5+H7亚型 三价灭活疫苗 抗原变异株 免疫效力 

分 类 号：S852.65[农业科学—基础兽医学]