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目的探讨pFLAG CMV8 gp96NTD-CSP重组DNA疟疾疫苗免疫能否诱导小鼠产生保护性免疫及其效应机制。方法以pFLAG CMV8质粒为载体,构建免疫用重组质粒,按照DNA疫苗免疫方法免疫小鼠;野生子孢子进行攻击后,采用Real-time PCR和吉氏染色观察被攻击小鼠的肝脏虫荷和原虫血症,即免疫小鼠抵御野生子孢子攻击的能力;并通过ELISA和ELISPOT方法探讨免疫小鼠保护性免疫的可能机制。结果核酸疫苗pFLAG CMV8 gp96NTD-CSP免疫小鼠能显著抵御野生子孢子的攻击,并且能诱导小鼠产生较高的抗体水平和较高的CSP特异的CD8+T细胞频率。结论 pFLAG CMV8 gp96NTD-CSP重组DNA疫苗可能通过诱导小鼠CSP特异抗体和CSP特异的CD8+T细胞的产生,一定程度上抵御野生子孢子的攻击。
Objective To investigate whether pFLAG CMV8 gp96NTD-CSP recombinant DNA vaccine can induce protective immunity in mice and its mechanism. Methods The plasmid pFLAG CMV8 was used as the vector to construct the recombinant plasmids for immunization and the mice were immunized with DNA vaccine. After wild sporozoites were challenged, the liver worms and protozoa of challenge mice were observed by Real-time PCR and Giemsa staining Serum, the ability of immune mice to resist the challenge of wild sporozoites; and explore the possible mechanism of protective immunity in mice immunized by ELISA and ELISPOT methods. Results The DNA vaccine pFLAG CMV8 gp96NTD-CSP could significantly resist the challenge of wild sporozoites and induce higher antibody production and higher CSP-specific CD8 + T cell frequency in mice. Conclusion The pFLAG CMV8 gp96NTD-CSP recombinant DNA vaccine may resist the challenge of wild sporozoites to a certain extent by inducing the production of mouse CSP-specific antibodies and CSP-specific CD8 + T cells.