HSV-2gD、gBCTL表位及Hsp70融合DNA疫苗的免疫效果观察

The immune responses of the DNA vaccine of HSV-2gD, gBCTL and Hsp70

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。

This study aim to evaluate humoral and cellular immune response in Balb/c mice immunized by pc （pcDNA3.1）-P6-gBCTL-Hsp70 genetic vaccine. Firstly the recombinant plasmid pc-P6-gBCTL-Hsp70 was transformed into CHO cells and the expressed protein was characterized by indirect immunofluorescence and Western blotting. Then the recombinant plasmid was inoculated into Balb/e mice by intramuscular injection for three times, once of two weeks. One week after the last vaccination, the levels of specific IgG antibody, IFN-γ and IL-18 were measured by ELISA; one month after the last vaccination, spleen cells of vaccinated mice were separated and the proliferation of spleen lymphoeytes was measured by MTT. We found that pc-P6-gBCTL-Hsp70 vaccine could promote splenocytes proliferation, induce higher levels of IFN-γ IL-18 and specific antibody, improve the activity of CTL kill cells of Balb/c mice. In conclusion, the recombinant clone pc-P6-gBCTL-Hsp70 can effectively induce humoral and cellular immune responses, which can offer basic conditions for constructing new type of polygenic DNA vaccine.