H5N1人用禽流感疫苗免疫原性检测方法的建立

目的确定人用禽流感疫苗免疫原性检测方法。方法用禽流感病毒R1203株制备疫苗,以不同剂量免疫家兔,检测血清中的血凝抑制抗体和中和抗体,并进行交叉血凝抑制试验、交叉单向免疫扩散试验和交叉中和试验。结果R1203株疫苗接种家兔1针后14 d,中和抗体和血抑抗体滴度均较低;第2针后14 d均明显升高。血抑抗体量-效反应不明显,而中和抗体量-效反应明显。交叉血凝抑制试验显示,异型之间普遍有交叉反应,滴度大部分不低于1∶40。单向免疫扩散试验显示异型之间无交叉反应。禽流感疫苗抗血清不能中和人流感病毒,但能中和同型病毒(R1194),滴度可达1∶240。结论中和抗体能正确反映疫苗的免疫原性;检测中和抗体的方法可用于禽流感疫苗的效力试验。

Development of A Method for Determination of Immunogenicity of H5N1 Avian Influenza Vaccine for Human Use

Objective To develop a method for determination of immunogenicity of avian influenza vaccine for human use.Methods Prepare vaccine with avian influenza virus strain R1203.Immunize rabbits with the prepared vaccine at various dosages and determine the HI and neutralizing antibodies in sera.Perform cross HI test,cross single immunodiffusion test and cross neutralization test on antisera against human and avian influenza viruses.Results Both neutralizing and HI antibody titers of rabbits were low 14 d after the 1st dose,but increased significantly 14 d after the 2nd dose.The increase of HI antibody titer was dose-independent,while that of neutralizing antibody was significantly dose-dependent.Cross HI test showed cross reaction of different types,and the HI titers of most of samples were not less than 1∶40.Cross single immunodiffusion test showed no cross reaction of antisera of different types.The antisera induced by avian influenza virus could not neutralize human influenza virus but could neutralize the virus of the same type(R1194),with the highest neutralizing titer of 1∶240.Conclusion Neutralizing antibody titer could reflect the immunogenicity of avian influenza vaccine correctly,thus determination of neutralizing antibody might be used as the potency test on avian influenza vaccine.