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目的研究多表位重组DNA疫苗pc(pcDNA3.1-)-P6-gBCTL-Hsp70的免疫原性。方法将构建的真核表达质粒pcP6-gBCTL-Hsp70转染CHO细胞表达,采用间接免疫荧光和Western blotting检测P6抗原的表达情况。将重组质粒肌肉注射免疫接种Balb/c小鼠3次,每次间隔2周。末次免疫后1周采集眼眶静脉血,末次免疫后1月无菌分离小鼠脾脏。采用ELISA法检测小鼠血清特异性抗体滴度、IFN-γ及IL-18含量,MTT法测定脾淋巴细胞增殖率,LDH法检测CTL活性。结果重组质粒pc-P6-gBCTL-Hsp70具有一定的免疫原性,可刺激机体产生特异性抗体,诱导分泌较高水平的IFN-γ和IL-18,促进小鼠脾淋巴细胞的增殖,CTL杀伤病毒感染细胞活性增强。结论重组核酸疫苗pc-P6-gBCTL-Hsp70能诱导较强的细胞免疫和体液免疫。
Objective To study the immunogenicity of multi-epitope recombinant DNA vaccine pc (pcDNA3.1 -) - P6-gBCTL-Hsp70. Methods The eukaryotic expression plasmid pcP6-gBCTL-Hsp70 was transfected into CHO cells and the expression of P6 antigen was detected by indirect immunofluorescence and Western blotting. Balb / c mice were immunized intramuscularly with the recombinant plasmids for 3 times at intervals of 2 weeks. Orbital venous blood was collected one week after the last immunization, and the spleen of mice was aseptically separated from the last immunization in January. Serum specific antibody titers, IFN-γ and IL-18 levels were detected by ELISA. The proliferation rate of splenic lymphocytes was measured by MTT assay and the activity of CTL by LDH assay. Results The recombinant plasmid pc-P6-gBCTL-Hsp70 had certain immunogenicity, which could stimulate the body to produce specific antibodies, induce the secretion of higher levels of IFN-γ and IL-18, promote the proliferation of mouse splenic lymphocytes and kill CTL Viral infected cells increased activity. Conclusion The recombinant DNA vaccine pc-P6-gBCTL-Hsp70 can induce strong cellular immunity and humoral immunity.