HPV16L1-E7蛋白疫苗诱导小鼠产生抗体及细胞免疫应答的研究

Induction of specific antibody and cellular immune response of HPV16 L1E7 chimeric virus-like particles in mice

目的 :探讨嵌合性病毒样颗粒HPV1 6L1 E7蛋白疫苗接种于动物体内诱发体液和细胞免疫反应的能力。方法 :以本室构建的HPV1 6L1 E7嵌合蛋白疫苗腹腔注射免疫BALB/c小鼠 ,在初次免疫后 ,分别于第 3、4、5周采血ELISA检测特异性抗体的产生。并耳皮下注射HPV1 6L1蛋白 ,检测特异性DHT反应。在末次免疫 2周后 ,取小鼠脾细胞 ,用HPV 1 6L1 E7蛋白刺激 ,检测T细胞增殖反应及血清IFN γ水平。结果 :HPV1 6L1 E7蛋白疫苗免疫小鼠可有效产生HPV1 6L1抗体及特异性DTH反应 ,加强免疫后 ,抗体水平升高 ,两者水平均高于对照组 (P <0 .0 1 )。HPV1 6L1 E7嵌合蛋白疫苗免疫后取小鼠脾细胞 ,在特异性HPV1 6L1 E7蛋白抗原刺激下 ,特异性T淋巴细胞明显增殖。被免疫小鼠血清中出现高水平IFN γ。结论 :嵌合病毒样颗粒HPV1 6L1 E7蛋白可诱导小鼠产生高滴度抗体、HPV1 6L1特异性TDH参与的超敏反应及细胞免疫应答的细胞因子 。

Objective: To assess the ability of inducing humoral and cell mediated immune responses of HPV16 L1E7 chimeric virus like particles for immunization in mice. Methods: Chimeric virus like particles (CVLPs) constructed by fusing HPV16 E7 sequences into the C terminus of the viral L1 gene were used as the preventive and therapeutic HPV vaccine. Animals received the vaccine intraperitoneally, and were detected for antibodies with ELISA, DTH reaction and serum IFN γ level five weeks after the first immunization. Spleenic T cells extracted from experimental mice were stimulated with HPV16 L1 E7 protein in vitro to show the activity of T lymphocytes two weeks after the second immunization. Results: Animals that received the cVLPs vaccine intraperitoneally developed systemic anti L1 IgG antibodies. And the level of antibodies was much higher after the second immunization. HPV16L1 specific DTH responses were induced by immunization with this vaccine. Serum IFN γ was much higher than that of the controls. Spleenic T cells extracted from experimental mice showed a specific T lymphocyte activity. Conclusion: The results suggest that HPV16 L1 E7 based protein vaccine may be useful for the prevention and therapy of primary infections by HPV16 and HPV associated tumor.