近交系小鼠H-2基因与接种乙型肝炎疫苗后无(弱)应答的相关性分析  

Correlation between H-2 gene of inbred strains of mice and no response or hyporesponse to hepatitis B vaccine

作　　者：马丽颖[1] 钟熙[1] 梁争论[1] 岳秉飞[1] 贺争鸣[1] 刘双环[1] 

机构地区：[1]中国食品药品检定研究院,北京100050

出　　处：《中国生物制品学杂志》2017年第7期678-684,共7页Chinese Journal of Biologicals

摘　　要：目的分析近交系小鼠H-2基因与接种乙型肝炎疫苗后无(弱)应答的相关性,建立免疫无应答小鼠动物模型,从基因水平和细胞因子水平研究免疫无应答的遗传机制,为乙肝疫苗的研制及人类免疫无应答的研究提供理论依据。方法采用微量细胞毒法检测不同近交系小鼠(DBA/1、BALB/c、NIH、SJL、C57BL/6、C57BL/10)的H-2单倍型;PCR法检测小鼠的H-2A和H-2E基因。用3种重组乙型肝炎疫苗(酿酒酵母、汉逊酵母、CHO细胞)和治疗性乙型肝炎疫苗(乙克)分别免疫6种不同品系近交系小鼠,采用ELISA法检测抗体水平,流式细胞术检测小鼠CD4^+、CD8^+、CD19^+T细胞和Treg细胞免疫前后的变化。结果不同近交系小鼠有不同的H-2单倍型。除封闭群小鼠NIH外,近交系小鼠SJL在A基因区内有两条200和500 bp的区带,其余均为1条区带;不同品系小鼠在H-2E基因区段均无异常表现。C57BL/10小鼠对乙肝疫苗表现为免疫无(弱)应答。结论 C57BL/10小鼠可作为免疫无应答实验动物模型,进行乙肝疫苗免疫无(弱)应答的相关性研究。建议在进行疫苗效力检测时先检测所用小鼠的H-2基因型,根据小鼠H-2基因的单倍型与免疫反应的相关性筛选实验用小鼠。Objective To analyze the correlation between H-2 gene of inbred strains of mice and no response or hyporesponse to hepatitis B（HB） vaccine, establish a mouse model of no response, investigate the genetic mechanism of no immune response at gene and cytokine levels and provide a theoretical basis for preparation of HB vaccine and study on no immune response in humans. Methods The H-2 haplotypes of various inbred mouse strains（DBA/1, BALB/c,NIH, SJL, C57BL/6 and C57BL/10） were determined by microcytotoxicity test, while the H-2A and H-2E genes by PCR. The six mouse strains were immunized with three recombinant HB vaccines（Saccharomyces cerevisiae, Hansenula polymorpha and CHO cells） and one therapeutic HB vaccine separately, of which the antibody levels were determined by ELISA, and the changes of CD4＋, CD8＋, CD19＋T cells and Treg cells before and after immunization by flow cytometry.Results H-2 haplotypes were different in various mouse strains. Two bands at lengths of 200 and 500 bp were observed in A gene regions of NIH and SIL mice, while a single band in other inbred mouse strains. No abnormalities were observed in H-2E gene regions of various mouse strains. However, C57BL/10 mice showed no response or hyporesponse to HB vaccine. Conclusion C75BL/10 mice may be used as a experimental model for study on no response or hyporesponse to HB vaccine. It is suggested that the H-2 genotype should be tested at first in vaccine potency test, and the experimental mice should be screened according to the correlation between haplotype of H-2 gene and immune response.

关 键 词：近交系小鼠 乙型肝炎疫苗 免疫无应答 H-2基因 

分 类 号：R373.21[医药卫生—病原生物学] R392-33[医药卫生—基础医学]