乙型肝炎疫苗细胞免疫应答检测用参考品的制备和初步应用研究

目的通过研究重组乙型肝炎表面抗原（HBsAg）免疫小鼠诱导细胞免疫应答的特点，制备一批细胞免疫应答检测用参考品，规范乙型肝炎疫苗细胞免疫实验条件和操作。方法背部皮下分别免疫小鼠（BALB／c，H-2^d）0．5、1、2、4和8鹏的HBsAg，于免后7d，检测小鼠脾单个核细胞（MNC）体外刺激诱生IFN-γ的水平。分别免疫小鼠2、4μgHBsAg，免后7d，检测MNC分泌IFN-γ水平。结果1μg组可检测小鼠IFN-μ阳转；免疫剂量与IFN-γ阳转率存在剂量效应关系（r=0．951，P=0．049），IFN-γSFC与免疫剂量也存在剂量效应关系（r=0．996，P=0．000）。2μgHBsAg组IFN-1阳转率在50％-75％之间；4μg组IFN-γ阳转率均为75％。结论制备一批乙肝疫苗细胞免疫参考品，用于规范疫苗诱导细胞免疫应答的方法检测。

Preparation and Preliminary Study on the Reference of Cellular Immune Response to Hepatitis B Vaccine

Objective To explore the character of cellular immune response induced by recombinant hepatitis B surface antigen and to prepare reference of cellular immune response to hepatitis B vaccine. Methods At different time, with different dose and by different person, the levels of IFN -γ secreted by splenic mononuclear cells （MNC） of mice were detected by ELISPOT after stimulation with HBsAg MHC class Ⅰ peptide S28-39. Results The positive conversion of IFN - γ was detected at 1μg dose of HBsAg. A dose - effect relationship between IFN - γ level and HBsAg dose was found （ r = 0. 951, P = 0. 049）. By different person and at different time, the positive conversion rates of IFN - γ varied from 50 % to 75 % with 2 μg of HBsAg and was 75 % with 4 μg of HBsAg respectively. Conclusion A batch of cellular immune references of Hepatitis B vaccine were prepared to standardize the cellular immune assay.