Epstein Barr病毒LMP1 DNA免疫小鼠免疫效果的测定

Evaluation of Immunization Effect in Epstein-Barr Virus LMP1 DNA Immunized Mice

【目的】探讨EBV LMP1重组体DNA免疫小鼠后的免疫效果。【方法】将已构建的EBVLMP1基因 (EBVLMP1)重组质粒 pcDNA3 BLMP1肌肉注射免疫Balb/c小鼠 ,在免疫后的不同时间分别用ELISA、流式细胞仪及LDH法检测免疫小鼠体内特异性EBVLMP1抗体滴度、CD4+ /CD8+ T淋巴细胞亚群的数量变化以及EBVLMP1特异性细胞毒性T淋巴细胞 (CTL)的活性。用 pcDNA3质粒及PBS作为阴性对照及空白对照。【结果】重组质粒免疫Balb/c小鼠后 ,可在外周血液血清中检测到特异性的EBVLMP1抗体 ;其脾细胞中CD4+ /CD8+ T淋巴细胞亚群数量明显高于阴性对照及空白对照 ,且可检测到EBVLMP1特异性的CTL存在。以上结果在免疫 16周之内无明显改变。【结论】EBV LMP1质粒重组体可刺激小鼠产生特异性的体液和细胞免疫 ,并且可维持一定的时间 ,具有较好的免疫原性 ,是一种有效、制备方便的EBV候选疫苗。

To observe the immune responses to the recombinant of Epstein Barr virus (EBV) LMP1 in mice by DNA immunization. An eukaryotic expression plasmid vector (pcDNA3) was engineered to contain EBV LMP1 gene (EBV LMP1). The recombinants were injected into the crural quadricep muscles of each Balb/c mice with 50 g/100 L plasmid DNA 50 μL. Blank vectors and PBS buffer were used as control. The antibody specific for EBV LMP1 and the percentage of CD4 +/CD8 + were detected by ELISA and flow cytometry at the different time after the plasmid DNA inoculation. Specific EBV LMP1 cytotoxic T lymphocytes (CTL) in the immunized mice spleen was detected by the lactate dehydrogenase (LDH) method. The specific EBV LMP1 antibodies can be detected in the sera of mice immunized with pcDNA3 BLMP1 and the percentage of CD4 +/CD8 + T lymphocytes derived form pcDNA3 BLMP1 immunized mice was obviously higher than that in the controls. The specific EBV LMP1 CTL could also be detected in recombinants inoculated mice but not in the controls. These results were observed up to 16 weeks after immunization. [Conclusion] The EBV LMP1 can induce the specific hormonal and cellular immune responses in the mice which can maintain for a period of time.