淋病奈瑟菌NspA基因疫苗诱导小鼠特异性体液免疫应答
首发时间:2007-04-11
摘要:目的 在构建淋病奈瑟菌NspA真核细胞表达质粒pcNspA 的基础上,进一步研究NspA基因疫苗诱生特异性抗体的作用。方法 将NspA真核表达质粒pcNspA通过肌肉注射、滴鼻免疫、阴道接种三种不同途径免疫BALB/c小鼠,收集免疫小鼠的血清、支气管肺泡洗液和阴道洗液,以间接ELISA方法检测抗体的类型和效价。比较不同免疫途径诱生小鼠体内特异性抗体水平的差异,并对小鼠产生的特异性抗体进行体外溶菌活性的检测。结果 滴鼻免疫和肌肉注射pcNspA质粒都可以在小鼠体内诱导产生较高的特异性体液免疫水平,尤其是滴鼻免疫途径不仅可诱生较高水平的血清IgG抗体(1:1600),而且在支气管肺泡洗液和阴道洗液中均可检测到特异性sIgA的存在。而阴道接种pcNspA质粒仅能在阴道局部检测到较低水平的特异性sIgA。用pcNspA质粒诱生的血清IgG抗体做体外溶菌活性实验,证实滴鼻免疫小鼠血清具有溶菌活性。结论 淋病奈瑟菌NspA基因疫苗可在小鼠体内诱导产生特异性抗体,并对机体有免疫保护作用;滴鼻免疫可能是淋病奈瑟菌NspA基因疫苗免疫的一个有效途径。
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Neisseria. gonorrhoeae specific antibodies in vivo in mice induced by NspA DNA vaccine and the bactericidal activity of the antiserum samples in vitro
Abstract:Objective To explore the feasibility of DNA vaccine against gonorrhea by using the self-construced pcNspA containing the NspA gene from Neisseria gonorrhoeae strain WHO-A. Methods BALB/c mice were immunized via intramuscular injection (IM), intranasal immunization (IN) or vaginal immunization (IV), respectively. Serum samples, lung lavage fluids and vaginal fluids were collected from each mouse 2 weeks after the last immunization. N. gonorrhoeae specific antibody levels in serum samples, pooled lung lavage fluids and vaginal wash fluids were analyzed by ELISA. Results All immunized mice generated measurable specific antibodies, and the antibody level in the IN mice was the highest. IN mice generated NspA-specific IgG and IgA in serum, as well as sIgA in lung lavage fluids and vaginal fluids. Antibody-mediated complement bactericidal activity assay in vitro indicated that the serum samples from the IN mice could mediate complement to produce bactericidal effect to N. gonorrhoeae. Conclusion These experimental results demonstrate that NspA DNA vaccine of pcNspA could induce protective immunity against N. gonorrhoeae infection and intranasal immunization was a suitable inoculation route with potential application value.
Keywords: Neisseria gonorrhoeae NspA DNA vaccine mouse
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