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目的：观察重组质粒ＤＮＡ直接免疫接种诱导ＢＡＬＢ／ｃ小鼠的免疫应答水平，为恶性疟原虫ＤＮＡ疫苗在动物和人体的应用提供依据。方法：构建编码多价保护性抗原的重组质粒ｐｃＤＮＡ３－Ｐｆ８，ＰＣＲ法检测免疫鼠的肌肉、肝脏、肾脏、心脏、脾脏和肺组织中ｐｃＤＮＡ３－Ｐｆ８，ＥＬＩＳＡ、Ｔ淋巴细胞转化试验、体外抑制试验观察其诱导的体液免疫及细胞免疫水平。结果：用ＰＣＲ法从上述组织中均检测到ｐｃＤＮＡ３－Ｐｆ８，ＥＬＩＳＡ法测得免疫鼠血清的特异性抗体滴度达１２５６０，淋巴细胞转化试验显示恶性疟原虫可溶性抗原能特异性地剌激免疫鼠脾细胞增殖，免疫血清在体外还能抑制恶性疟红内期疟原虫的生长、发育。结论：编码恶性疟原虫多价保护性抗原的重组质粒ｐｃＤＮＡ３－Ｐｆ８直接免疫接种，能特异性地剌激ＢＡＬＢ／ｃ小鼠产生体液免疫和细胞免疫应答，其免疫血清在体外对疟原虫生长发育具有明显抑制作用。
OBJECTIVE: To observe the level of immune response in BALB / c mice induced by direct DNA immunization of recombinant plasmids and provide a basis for the application of Plasmodium falciparum DNA vaccine in animals and humans. Methods: The recombinant plasmid pcDNA3-Pf8 encoding multivalent protective antigen was constructed. The pcDNA3-Pf8, ELISA, T lymphocyte transformation test in muscle, liver, kidney, heart, spleen and lung of immunized mice were detected by PCR. Observe the induced humoral and cellular immunity. Results: The pcDNA3-Pf8 was detected by PCR from all the above tissues. The titer of the specific antibody of the immunized rat serum was 12560 measured by ELISA, and the lymphocyte transformation test showed that the soluble antigen of P. falciparum could specifically stimulate the immunized rat Spleen cell proliferation, immune serum in vitro can inhibit Plasmodium falciparum malaria parasite growth and development. CONCLUSION: Direct immunization of recombinant plasmid pcDNA3-Pf8 encoding P. falciparum multivalent protective antigen can specifically stimulate the humoral and cellular immune response in BALB / c mice. The immune serum of P. falciparum is able to stimulate the growth and development of Plasmodium in vitro Has a significant inhibitory effect.