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目的 初步探讨HIV-1CN54合成gp120基因的DNA疫苗（pcDNA3.1-syngp120）鼻内接种小鼠是否诱发免疫应答。方法 DNA疫苗免疫后,制备脾和肠系膜淋巴结（MLN）淋巴细胞,在体外测其增殖应答和CD8+CTL应答。间接ELISA法测血清和粘膜洗液抗原-特异的IgG和IgA抗体滴度。中和实验测免疫血清和阴道洗液是否中和 HIV-1SF33。结果 在 DNA疫苗未次免疫后,小鼠第 1周检测到脾和 MLN CD8+CTL应答较弱,而第 5周未检测到。另外,在末次免疫后的第1、5周检测到MLN而未检测到脾淋巴细胞发生增殖应答。并且检测到特异的血清IgG抗体和粘膜的（包括粪便和阴道洗液）IgA抗体,但未检测到血清的IgA抗体和粘膜的（包括粪便和阴道洗液）IgG抗体。中和实验发现末次免疫后第5周的血清能中和实验室毒株HIV-1SF33（B亚型）,而阴道洗液则没有。结论 该DNA疫苗鼻内免疫小鼠可诱导粘膜免疫应答,包括 MLN淋巴细胞增殖应答和 CD8+CTL应答,同时诱导较弱的粘膜IgA抗体应答。此外,能诱导脾CD8+CTL应答和血清IgG抗体应答。免疫血清中和HIV-1S F33,而阴道洗液不能。
Objective To investigate whether the intranasal vaccination with pcDNA3.1-syngp120 DNA vaccine of gp120 gene of HIV-1CN54 induces an immune response. Methods After spleen and mesenteric lymph node (MLN) lymphocytes were immunized with DNA vaccine, their proliferative response and CD8 + CTL response were measured in vitro. Indirect ELISA was used to measure serum and mucosal wash antigen-specific IgG and IgA antibody titers. Neutralization test to determine whether immune serum and vaginal lotion neutralize HIV-1SF33. Results After the mice were immunized with no DNA vaccine, the response to spleen and MLN CD8 + CTL was weak in the first week and not detected in the fifth week. In addition, MLNs were detected at weeks 1 and 5 after the last immunization and no proliferative response was detected in splenic lymphocytes. And specific serum IgG antibodies and mucosal IgA antibodies (including stool and vaginal washes) but no serum IgA and mucosal IgG antibodies (including stool and vaginal washes) were detected. Neutralization experiments revealed that serum 5 weeks after the last immunization neutralized laboratory strain HIV-1SF33 (subtype B), whereas vaginal washings did not. Conclusion The intranasal immunization of the DNA vaccine induced mucosal immune responses, including MLN lymphocyte proliferation and CD8 + CTL responses, while inducing a weaker mucosal IgA antibody response. In addition, splenic CD8 + CTL responses and serum IgG antibody responses can be induced. Immune serum neutralizes HIV-1S F33, whereas vaginal lotion does not.