摘要
为获得用于评价H7N9亚型禽流感疫苗诱导的抗体Fc免疫效应的靶细胞,研究采用慢病毒包装与感染技术构建一株稳定表达H7N9亚型禽流感病毒(H7N9 AIV)HA蛋白的细胞株,并以该细胞株为靶细胞建立H7N9亚型禽流感疫苗免疫血清补体介导的细胞毒(ADCML)活性的检测方法。结果显示成功包装了一株重组慢病毒,滴度可达108 FU/mL;构建了稳定表达H7N9 AIV HA蛋白的293T细胞株,HA蛋白主要在细胞膜上表达。基于新城疫病毒(NDV)载体的H7N9亚型禽流感疫苗免疫鸡后,诱导产生的HA结合性IgG抗体滴度为2180。以稳定细胞株为靶细胞,载体疫苗血清诱导的细胞死亡水平显著高于空载体与未免疫鸡血清(P<0.001);补体灭活后,载体疫苗血清诱导的细胞死亡水平显著降低(P<0.001)。研究表明,基于NDV载体的H7N9亚型禽流感疫苗免疫鸡血清具有较强的ADCML活性,稳定表达H7N9 AIV HA蛋白的细胞株可作为评价H7N9亚型禽流感疫苗免疫血清Fc免疫效应的靶细胞。
This study aimed to generate a cell line as the target cells for assessing Fc effector functions of serum against H7N9 subtype avian influenza vaccines.Lentivirus packaging and infection technology was used to generate a cell line that could stably express the haemuglutinin(HA)of H7N9 subtype avian influenza virus(H7N9 AIV).Using the stable cell line as the tar⁃get cells,a method measuring antibody-dependent complement-mediated lysis(ADCML)activity of H7N9 subtype avian influen⁃za vaccine immune serum was established.A recombinant lentivirus with a high virus titer(108 fluorescence unit/mL)was pack⁃aged and a stable 293T cell line expressing the HA protein of H7N9 AIV mainly on the cell surface was generated through lentivi⁃rus infection.Mean HA-binding IgG titer of the sera from chickens immunized with the Newcastle disease virus(NDV)-vectored H7N9 subtype avian influenza vaccine was 2180.These sera caused a significantly higher level of cell death of the stable 293T cells compared to the vector and mock-control chicken sera(P<0.001).In addition,after complement inactivation,the level of cell death induced by the NDV-vectored H7N9 subtype avian influenza vaccine sera significantly decreased(P<0.001).It suggested that chicken sera induced by the NDV-vectored H7N9 subtype avian influenza vaccine could elicit a strong H7N9-specific ADC⁃ML activity,and the cell line stably expressing the HA protein could be used as the target cells to determine Fc effector functions of H7N9 subtype avian influenza vaccine immune sera.
作者
曹永忠
邓婧
黄雅
薄宗义
胡增垒
CAO Yongzhong;DENG Jing;HUANG Ya;BO Zongyi;HU Zenglei(Joint International Research Laboratory of Agriculture and Agri-Product Safety,Ministry of Education,Yangzhou University,Yangzhou,Jiangsu 225009;Animal Infectious Disease Laboratory,Ministry of Agriculture and Rural Affairs,Yangzhou University,Yangzhou,Jiangsu 225009;Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses,Yangzhou University,Yangzhou,Jiangsu 225009;Jiangsu Key Laboratory of Zoonosis,Yangzhou University,Yangzhou,Jiangsu 225009)
出处
《中国家禽》
北大核心
2023年第10期59-64,共6页
China Poultry
基金
江苏省农业科技自主创新资金项目(CX(21)3129)
江苏高校优势学科建设工程资助项目(PAPD)。
关键词
H7N9亚型禽流感
HA蛋白
稳定细胞株
疫苗
Fc免疫效应
补体
抗体保护
H7N9 subtype avian influenza
HA protein
stable cell line
vaccine
Fc effector functions
complement
anti⁃body protection
