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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 relationship between H-2 gene in inbred mice and no (weak) response after inoculation of hepatitis B vaccine and to establish an animal model of immune nonresponsive mice and to study the genetic mechanism of immune non-response from gene level and cytokine levels , Provide a theoretical basis for the study of hepatitis B vaccine and human immune non-response. Methods The H-2 haplotypes of different inbred mice (DBA / 1, BALB / c, NIH, SJL, C57BL / 6 and C57BL / 10) 2A and H-2E genes. Six different strains of inbred mice were respectively immunized with three recombinant hepatitis B vaccines (Saccharomyces cerevisiae, Hansenula polymorpha, CHO cells) and therapeutic hepatitis B vaccine (Bg), and the antibody levels were detected by ELISA The changes of CD4 ~ +, CD8 ~ +, CD19 ~ + T cells and Treg cells in mice before and after immunization were detected by the cell cytometry. Results Different inbred mice had different H-2 haplotypes. In addition to the closed group mice NIH, inbred mice SJL in the A gene region has two 200 and 500 bp bands, the rest are a band; different strains of mice in the H-2E gene segments No abnormalities. C57BL / 10 mice appear immune-no (weak) response to hepatitis B vaccine. Conclusion C57BL / 10 mice can be used as immunological non-response animal model to study the correlation between non-immune response and non-immune response of hepatitis B vaccine. It is recommended to test the H-2 genotype of the mice used for vaccine efficacy testing and to screen the experimental mice based on the correlation between the H-2 haplotype and the immune response.