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目的 :探讨基于α 病毒的基因疫苗的免疫学效应作用 ,寻找更好的基因疫苗形式。方法 :用磷酸钙法共转染表达质粒P1A/pSMART2a和包装质粒helper到 2 93细胞中 ,制备高水平的重组α 病毒P1A/SFV。鉴定病毒及其表达后 ,用其免疫小鼠 ,测定免疫动物体内抗体生成情况和特异CTL的杀伤效应。在预防性和治疗性实验中 ,测定该重组病毒免疫动物后6 0d内动物的无瘤率和生存率。结果 :该重组病毒疫苗在体外培养细胞中有很好的表达。用其免疫动物后 ,P1A/SFV能激发比对照组强得多的CTL杀伤效应。在E/T比为 10 0∶1时 ,P1A/SFV组CTL的杀伤率为 75 %。抗体在所有组别中均未见产生。在保护性和治疗性实验中 ,于 6 0d实验期限内 ,P1A/SFV的效果最好。保护性实验第 6 0天时 ,P1A/SFV组动物的无瘤率达到 6 0 % ;而P1A/ pCI neo组只有2 0 %。在治疗性实验第 6 0天 ,P1A/SFV组小鼠的存活率达到 5 0 %左右 ;而其他对照组都只有 10 %左右。结论 :相对于普通基因疫苗 ,基于重组α 病毒的基因疫苗效果最好 ,为临床肿瘤的基因治疗提供了新的思路
OBJECTIVE: To investigate the immunological effects of alphavirus-based gene vaccines and to search for better gene vaccine forms. Methods: The recombinant adenovirus P1A / SFV was prepared by co-transfection of the expression plasmid P1A / pSMART2a and the packaging plasmid helper into 293 cells by the calcium phosphate method. After identification of the virus and its expression, the mice were immunized and the antibody production in the immunized animal and the killing effect of the specific CTL were measured. In preventive and curative experiments, the tumor-free rate and survival rate of the animals within 60 days after the recombinant virus-immunized animal were determined. Results: The recombinant virus vaccine in vitro cultured cells have a good expression. After being immunized with the same, P1A / SFV stimulated a much stronger CTL killing effect than the control group. At the E / T ratio of 10: 1, the killing rate of CTL in the P1A / SFV group was 75%. Antibodies were not found in all groups. In protective and therapeutic experiments, P1A / SFV performed best during the 60d experimental period. On the 60th day of protective experiment, the tumor-free rate of animals in P1A / SFV group was 60%, while that in P1A / pCI neo group was only 20%. On the 60th day of the therapeutic experiment, the survival rate of mice in the P1A / SFV group was about 50%, while that in the other control groups was only about 10%. CONCLUSION: Gene vaccines based on recombinant alphavirus have the best effect compared with common gene vaccines, providing new ideas for gene therapy of clinical tumors