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目的 研究GM CSF(粒 单核巨噬细胞集落刺激因子 )与抗原化抗体联合基因免疫时 ,GM CSF对TH 细胞应答的调节作用。方法 在编码免疫球蛋白重链的质粒中分别克隆黏蛋白 1(MUC1 )中特异性PDTRP抗原表位和GM CSF编码基因 ,构建含PDTRP和GM CSF编码基因的抗原化抗体表达重组体。经脾免疫和肌肉加强的方式免疫BALB c小鼠。采用ELISA方法检测小鼠血清中特异性抗体的水平及其亚类并动态观察 ;RT PCR方法检测TH 细胞分化中细胞因子和转录因子的表达水平。结果 抗原化抗体基因免疫能诱导机体产生免疫应答。GM CSF增强抗原化抗体基因免疫诱生的抗PDTRP特异性IgG抗体水平并且伴随IgG1 IgG2a显著性升高 ,同时可增强淋巴细胞TH2型细胞因子及转录因子 (IL 4、GATA 3)mRNA的表达水平。结论 GM CSF在增强抗原化抗体基因免疫诱导的免疫应答的同时使得免疫应答向TH2方向偏移。
Objective To study the regulatory effect of GM CSF on TH cell responses in combination with GM CSF (granulocyte-macrophage colony-stimulating factor) and antigen-binding antibody. Methods The specific PDTRP epitopes and GM CSF encoding genes in mucin 1 (MUC1) were cloned into the immunoglobulin heavy chain-encoding plasmids respectively to construct recombinant antigen-antibody expression plasmids containing PDTRP and GM CSF coding genes. BALBc mice were immunized spleenically and muscularly. The levels of specific antibodies and their subclasses in serum were detected by ELISA. The expression of cytokines and transcription factors in TH cells were detected by RT-PCR. Results Antigenized antibody gene immunity can induce the body to produce immune response. GM CSF enhances the level of anti-PDTRP-specific IgG antibodies induced by gene immunization of antigen-presenting cells with a significant increase of IgG1 IgG2a, as well as enhances the expression of TH2-type cytokines and transcription factors (IL4, GATA3) mRNA in lymphocytes . Conclusion GM CSF enhances the immune response induced by antigen immunization of antigen and at the same time shifts the immune response towards TH2.