重组痘苗病毒HIV疫苗肌注初免蛋白疫苗滴鼻加强后激发高强度黏膜IgA应答反应

摘要

Objective To analyze the mucosal immune responses induced in BALB/c mice after immunization with Vaccinia Tiantan-based HIV vaccine in combination with protein and to evaluate the effi-cacy of different immune strategies and adjuvants.Methods The BALB/c mice were intramuscularly or in-tranasally immunized with the recombinant Vaccinia virus Tiantan strain ( rTV) carrying CN54 Gag-Pol-Env gene and boosted with the gp140 protein and MF59 adjuvant by intramuscular, intranasal or subcutaneous in-jection.Serum, saliva and vaginal lavage samples were collected from the BALB/c mice.The titers of anti-gen specific IgG and IgA were detected by ELISA.Results Significantly enhanced mucosal immune respon-ses were induced by immunization with gp140 protein used in combination with MF59 adjuvant.The IgA an-tibodies elicited in mice mucosal tissues by gp140 protein and MF59 adjuvant were as high as those induced by gp140 protein and cholera toxin subunit B or recombinant flagellin adjuvant.High tiers of gp140-specific IgA antibodies were observed in serum, saliva and vaginal lavage samples from the mice intramuscularly primed with rTV and intranasally boosted with gp140 protein and MF59 adjuvant.The tiers of IgA antibodies in serum and mucosal tissue samples were 1 ∶15 000 and 1 ∶600, respectively.Conclusion High titers of mucosal IgA antibodies were elicited in BALB/c mice intramuscularly primed with Vaccinia Tiantan-based HIV vaccine and intranasally boosted with gp140 protein and MF59 adjuvant, especially in vaginal and oral tissues.This immunization strategy might be able to block the heterosexual transmission of HIV-1 through va-ginal and oral mucosa.%目的：研究痘苗病毒载体HIV疫苗和蛋白疫苗联合运用所激发的黏膜免疫应答，探讨不同途径免疫产生黏膜免疫应答的异同，进一步证实MF59佐剂在激发疫苗黏膜免疫应答方面的效果。方法以表达HIV-1中国流行株 CN54 Gag-Pol-Env 基因的重组痘苗病毒天坛株rTV，重组HIV-1膜蛋白gp140三聚体为抗原，辅以水包油型佐剂（ oil-in-water） MF59，在小鼠模型上，通过不同免疫途径组合联合免疫BALB／c小鼠，即重组痘苗病毒初免，免疫途径为滴鼻或肌注，gp140与MF59混合物加强免疫，免疫途径为滴鼻、肌注或皮下，探讨能诱导高效黏膜免疫应答的免疫策略。通过ELISA方法对小鼠血清中HIV-1特异性的IgG和IgA及唾液和阴道分泌物中IgA进行检测。结果MF59是一种良好的黏膜佐剂，能显著增强HIV蛋白疫苗的黏膜免疫应答，所激发的黏膜IgA滴度与鞭毛素蛋白（ SF）和霍乱毒素B亚单位（ CTB）的黏膜佐剂活性相当；重组痘苗病毒初免（肌注），gp140与MF59加强（滴鼻）联合免疫策略能诱导高水平的血清IgA和黏膜IgA抗体（P＜0．05），其抗体滴度分别高达1∶15000和1∶600。结论重组痘苗病毒天坛株rTV系统途径免疫小鼠后蛋白疫苗黏膜途径加强能激发高强度黏膜免疫应答，这一免疫策略及组合可为阻断通过口腔和阴道黏膜传播的艾滋病疫苗研究提供参考。