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目的用构建的pVAX-p55-v3DNA疫苗免疫SD大鼠,以pVAX-p55-v0作为阳性对照,从细胞及体液免疫水平研究其免疫保护作用机制。方法分别将pVAX-p55-v3、pVAX-p55-v0(阳性对照)DNA疫苗免疫SD大鼠(以pVAX1空载体及PBS作为阴性对照),然后按常规方法建立PC动物模型,于第6周处死大鼠,制作肺印片,计数包囊;取脾,做淋巴细胞增殖试验,测定各组大鼠脾T细胞增殖水平;取血,分离血清,采用ELISA法测定血清中细胞因子IFN-γ、IL-4、IL-2及抗体水平,评价疫苗的免疫水平就其免疫保护作用。结果与pVAX-p55-v0组一样,pVAX-p55-v3免疫组肺印片包囊数较PBS及pVAX1对照组明显减少[(1.75±0.98)个,P<0.05),脾T淋巴细胞显著增殖[(0.1937±0.0156),P<0.05),血清IFN-γ、IL-2水平增高明显[(74.279±2.215)pg/ml,(47.409±3.814)pg/ml,P<0.05),血清IgG显著增高[(1.22±0.09),P<0.05),而免疫下调因子IL-4水平各实验组之间无显著性差异(P>0.05)。结论重组DNA疫苗pVAX-p55-v3可诱导大鼠产生保护性免疫应答。其免疫保护效应与p55-v0DNA疫苗无显著差别。
OBJECTIVE: To construct a pVAX-p55-v3 DNA vaccine for immunization of SD rats and to investigate the mechanism of its immunoprotection based on cellular and humoral immunity using pVAX-p55-v0 as a positive control. Methods SD rats were immunized with pVAX-p55-v3 and pVAX-p55-v0 (positive control) DNA vaccine respectively, and pVAX1 empty vector and PBS were used as negative controls. PC animal models were established by routine methods and sacrificed at the 6th week Rats were sacrificed and the lungs were printed and counted. The splenic lymphocytes were obtained and the proliferation of splenic T lymphocytes was measured. The serum levels of splenic T cells were measured. The levels of serum IFN-γ, IL-4, IL-2 and antibody levels, evaluate the immune level of the vaccine on its immune protection. Results Compared with pVAX-p55-v0 group, the number of cytoplasm of lung imprint pVAX-p55-v3 group was significantly decreased compared with PBS group and pVAX1 control group [(1.75 ± 0.98), P <0.05) The levels of serum IFN-γ and IL-2 increased significantly ([74.279 ± 2.215] pg / ml, (47.409 ± 3.814) pg / ml, P <0.05) (1.22 ± 0.09, P <0.05), while there was no significant difference between the experimental group and the immunosuppressive factor IL-4 (P> 0.05). Conclusion The recombinant DNA vaccine pVAX-p55-v3 can induce protective immunity in rats. The immune protection effect and p55-v0 DNA vaccine no significant difference.