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目的 :检测HPV 18L1 E6、E7嵌合基因DNA疫苗在小鼠体内的体液和细胞免疫效应。方法 :将实验动物BALB/c小鼠 5 4只随机分为 9组 ,按不同免疫方式 (肌肉接种或鼻内滴注 )分别给予不同的重组质粒 (pVAX1 L1 E6M3或pVAX1 L1 E7M3)和免疫佐剂 (pLXHDmB7 2或LTB)。用免疫原免疫 3次 ,末次免疫后取眼球后血进行ELISA抗体检测。末次免疫后进行小鼠足垫迟发型超敏反应 (DTH)试验。断足进行小鼠足垫HE染色。取小鼠脾脏制成单细胞悬液 ,进行脾细胞增殖试验 ,并进行CD4 + /CD8+ T细胞中IFN γ+ 或IL 4 + 细胞的FACS分析。结果 :与对照组相比 ,各实验组均获得明显的免疫效果。实验组免疫后血清抗体A值均高于相应组别免疫前 ;肌肉注射组每次免疫后抗体水平较前次明显升高。实验组小鼠注射VLP抗原的左后足垫局部有红肿硬结 ,镜下观察可见大量单核细胞侵润。肌肉接种组的DTH反应强度、脾细胞增殖刺激指数 (SI)和CD8+ IFN γ+ 细胞数均高于鼻内滴注组 ;而鼻内滴注组CD4 + IL 4 + 细胞数高于单纯质粒肌肉接种组 ;加入pLX HDmB7 2的联合免疫组各项指标均高于单纯质粒组。结论 :证实了重组pVAX1 HPV18L1/E6、E7嵌和基因DNA疫苗能诱导小鼠的体液免疫和细胞免疫效应。同时 ,证实B7 2分子能显著提高该质粒在小鼠体内的免疫反应?
Objective: To detect the humoral and cellular immune responses of HPV 18L1 E6 and E7 chimeric DNA vaccines in mice. Methods: Five 4 BALB / c mice were randomly divided into 9 groups. Different recombinant plasmids (pVAX1 L1 E6M3 or pVAX1 L1 E7M3) and immune adjuvant were given by different immunizations (intramuscular or intranasal instillation) (PLXHDmB7 2 or LTB). Immunized with the immunogen three times, after the last immunization of the eyeball to take ELISA antibody detection. Mice footpad delayed-type hypersensitivity (DTH) test was performed after the last immunization. Broken foot mouse HE padding. The spleen of the mouse was made into a single cell suspension, and the spleen cell proliferation test was carried out, and the FACS analysis of IFN γ + or IL 4 + cells in CD4 + / CD8 + T cells was performed. Results: Compared with the control group, all the experimental groups were significantly immunized. The serum A value in the experimental group after immunization was higher than that of the corresponding group before immunization. The antibody level in the intramuscular injection group was obviously higher than the previous one. Experimental group of mice injected with VLP antigen left foot pad locally inflamed sclerosis, microscopic observation showed a large number of monocyte infiltration. The intramuscular inoculation group had higher DTH intensity, splenocyte proliferation stimulating index (SI) and CD8 + IFN γ + cell number than the intranasal instillation group, while the number of CD4 + IL 4 + cells in the intranasal instillation group was higher than that of the pure plasmid muscle Inoculation group; adding pLX HDmB7 2 combined immunization group indicators were higher than the plasmid group alone. Conclusion: The recombinant DNA vaccine of pVAX1 HPV18L1 / E6, E7 and DNA vaccine can induce humoral and cellular immune responses in mice. At the same time, it was confirmed that B7 2 molecule can significantly enhance the immune response of the plasmid in mice.