纳米佐剂N-2-HACC/CMCS对鸡新城疫病毒和H9禽流感病毒二联灭活疫苗免疫效果的评价  

Evaluation of immune enhancement of nano adjuvant N-2-HACC/CMCS on the combined inactivated vaccine against Newcastle disease virus and H9 subtype avian influenza virus

作　　者：赵志 高远 舒建洪[4] 孙德君 金政 赵凯 ZHAO Zhi;GAO Yuan;SHU Jian-hong;SUN De-jun;JIN Zheng;ZHAO Kai(Engineering Research Center of Agricultural Microbiology Technology,Ministry of Education,Heilongjiang University,Harbin 150500,China;Key Laboratory of Microbiology,College of Heilongjiang Province,School of Life Sciences,Heilongjiang University,Harbin 150080,China;Institute of Nanobiomaterials and Immunology,School of Life Science,Taizhou University,Taizhou 318000,China;College of Life Sciences and Medicine,Zhejiang Sci-Tech University,Hangzhou 310018,China;Harbin Pharmaceutical Group Bio-vaccine Co.Ltd,Harbin 150069,China;Zhejiang Hom-Sun Biotechnology Co.Ltd.,Shaoxing 312366,China)

机构地区：[1]黑龙江大学农业微生物技术教育部工程研究中心,黑龙江哈尔滨150500 [2]黑龙江大学生命科学学院黑龙江省普通高校微生物重点实验室,黑龙江哈尔滨150080 [3]台州学院生命科学学院/纳米生物材料与免疫学研究所,浙江台州318000 [4]浙江理工大学生命科学与医药学院,浙江杭州310018 [5]哈药集团生物疫苗有限公司,黑龙江哈尔滨150069 [6]浙江洪晟生物科技股份有限公司,浙江绍兴312366

出　　处：《中国预防兽医学报》2022年第6期630-635,共6页Chinese Journal of Preventive Veterinary Medicine

基　　金：浙江省“尖兵”“领雁”研发攻关计划(2022C02031);国家自然科学基金(31771000);哈尔滨市“雏鹰计划”暨科技企业培育专项(2020CYJBCG0306)。

摘　　要：为了评估纳米佐剂N-2-HACC/CMCS在鸡新城疫病毒(NDV)和H9禽流感病毒(AIV)二联灭活疫苗中的免疫效果,本研究以N-2-HACC/CMCS为佐剂制备了鸡NDV和AIV二联灭活疫苗(N-2-HACC/CMCS NDV+H9AIV二联灭活疫苗,简称为佐剂二联灭活疫苗),根据现行《中国兽药典》要求对制备的疫苗进行了外观、剂型、稳定性、无菌和安全性的检测后又评价了该疫苗的免疫效果。即将50只3日龄SPF鸡分为5组,分别为0.2 mL佐剂二联灭活疫苗组、0.2 mL商品二联灭活疫苗组、0.3 mL佐剂二联灭活疫苗组、0.3 mL商品二联灭活疫苗组及空白对照组,免疫后不同时间采血,采用血凝抑制试验和ELISA方法分别检测各组鸡血清中的NDV和AIV HI抗体效价以及细胞因子(IFN-γ、IL-2、IL-4)的分泌水平。并根据抗体检测结果分析免疫剂量与HI抗体效价和细胞因子的相关性。结果显示,无论是0.2 mL还是0.3 mL的免疫剂量,佐剂二联灭活疫苗免疫鸡血清中NDV和AIV HI抗体效价均极显著高于商品二联灭活疫苗组鸡(P<0.01);商品二联灭活疫苗免疫鸡产生的NDV和AIV HI抗体效价与免疫剂量呈正相关,而佐剂二联灭活疫苗免疫鸡产生的NDV和AIV HI抗体效价与免疫剂量无相关性;细胞因子检测结果显示,免疫后21 d,0.2 mL佐剂二联灭活疫苗和商品二联灭活疫苗免疫鸡血清中的IFN-γ、IL-2、IL-4含量均极显著高于各疫苗免疫后7 d和14 d各组鸡血清中的各细胞因子(P<0.01),而免疫后21 d,0.3 mL上述两种二联灭活疫苗免疫鸡血清中的细胞因子含量与免疫后7 d和14 d相应组鸡的细胞因子含量相比无明显升高,其中免疫0.2 mL佐剂二联灭活疫苗产生的IFN-γ、IL-2、IL-4水平最高。本研究结果首次证实N-2-HACC/CMCS不仅可以提高免疫鸡血清中NDV和AIV HI抗体效价,而且还可以减少疫苗中的抗原用量。本研究为N-2-HACC/CMCS作为疫苗佐剂在NDV和H9 AIV等灭活疫苗中的应用奠定了实验基础。In order to evaluate the immune effect of nano adjuvant N-2-HACC/CMCS in the combined inactivated vaccine against Newcastle disease virus(NDV)and H9 avian influenza virus(AIV),this study prepared a combined inactivated vaccine against NDV and H9 AIV with N-2-HACC/CMCS as adjuvant(N-2-HACC/CMCS NDV+H9 AIV combined inactivated vaccine,for short named adjuvant combined inactivated vaccine).In accordance with the current standard of the Chinese Veterinary Pharmacopoeia,the appearance,formulation,stability,sterility,and safety of the prepared vaccine were tested,and the immune effect of the vaccine was evaluated in chickens.503-day-old SPF chickens were divided into five groups,including 0.2mL adjuvantcombined inactivated vaccine group,0.2mL commercial combined inactivated vaccine group,0.3mL adjuvant-combined inactivated vaccine group,0.3mL commercial combined inactivated vaccine group and blank control group,respectively.After immunization,blood samples were collected at different time points.The hemagglutination inhibition(HI)antibody titers to NDV and AIV and the secretion levels of cytokines(including IFN-γ,IL-2,IL-4)in the serum of chickens in each group were detected by HI assay and ELISA,respectively,and the correlation between immune doses and HI antibody titers and cytokines was analyzed according to test results.The results showed that the HI antibody titers of NDV and AIV in the serum of chickens immunized with adjuvantcombined inactivated vaccine were significantly higher than those of chickens immunized with commercial combined inactivated vaccine at either 0.2mL or 0.3mL doses(P<0.01);the HI antibody titers agaisnt NDV and AIV in chickens immunized with commercial combined inactivated vaccine were positively correlated with the immune dose,while the HI antibody titers against NDV and AIV HI in chickens immunized with adjuvant combined inactivated vaccine were not correlated with the immune dose;the results of cytokine detection showed that the levels of IFN-γ,IL-2 and IL-4 in the serum of chickens imm

关 键 词：新城疫 H9亚型禽流感 壳聚糖季铵盐 免疫佐剂 免疫效果 

分 类 号：S852.4[农业科学—基础兽医学]