中华皮肤科杂志

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脂质体包裹HSV-2gD-HSP70融合蛋白诱导小鼠免疫应答的研究

樊建勇, 杨慧兰   

  1. 广州军区广州总医院皮肤科, 510010
  • 收稿日期:2006-11-29 出版日期:2007-10-15 发布日期:2007-10-15
  • 通讯作者: 杨慧兰,email:huilany@medmail.com.cn E-mail:huilany@medmail.com.cn
  • 基金资助:
    国家自然科学基金(30371290);广东省医学科研基金资助课题(A2006492)

A study on the immune response induced by liposome-entrapped HSV-2gD-HSP70 fusion protein in mice

FAN Jian-yong, YANG Hui-lan   

  1. Department of Dermatology, Guangzhou General Hospital of Guangzhou Military Area Command of Chinese PLA, Guangzhou 510010, China
  • Received:2006-11-29 Online:2007-10-15 Published:2007-10-15

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摘要： 目的 探讨局部外用脂质体包裹单纯疱疹病毒2型(HSV-2)糖蛋白D(glycoprotein D,gD)与结核杆菌热休克蛋白(heat shock protein,HSP)70融合蛋白疫苗在小鼠体内诱导产生特异性免疫应答的情况。方法 局部外用脂质体包裹gD,gD+HSP70,HSP70-gD融合蛋白分别免疫BALB/c小鼠3周,末次免疫后第4周检测小鼠血清中gD IgG水平,同时分离小鼠脾淋巴细胞,并用HSV-2gD蛋白刺激,用MTT法检测小鼠脾淋巴细胞增殖情况及ELISA法检测脾淋巴细胞培养上清中γ-干扰素(INF-γ)、白细胞介素4(IL-4)水平。结果 脂质体包裹gD、gD+HSP70和HSP70-gD均可以诱导小鼠产生针对HSV-2gD蛋白的抗体并诱导脾淋巴细胞增殖及分泌IFN-γ和IL-4,脂质体包裹HSP70-gD组在血清抗体水平、脾淋巴细胞刺激指数及其分泌IFN-γ、IL-4水平方面均显著高于脂质体包裹gD+HSP70组和gD组,差异有统计学意义(P<0.05)。结论 HSP70可以促进gD蛋白诱导小鼠特异性免疫应答反应。

关键词: HSP70热休克蛋白质类, 糖蛋白类, 单纯疱疹病毒疫苗, 脂质体

Abstract: Objective To investigate the specific immune response induced by topical application of liposome-entrapped herpes simplex virus-2 glycoprotein D(HSV-2gD)-Bacillus tuberculosis heat shock protein(HSP)70 fusion protein in mice.Methods Forty BALB/C mice were randomly divided into 4 groups to be immunized by empty liposomes,liposome-entrapped gD,liposome-entrapped gD and HSP70, and liposome-entrapped gD-HSP70 fusion protein,respectively,once a day for 3 weeks.At 4 weeks after the last immunization,the serum level of anti-HSV-2gD IgG antibody was assessed.At the same time,lympho-cytes were isolated from the spleen of the vaccinated mice and stimulated with HSV-2gD protein,the prolif-eration of lymphocytes was measured by MTT assay,and ELISA assay was utilized to measure the levels of IFN-γand IL-4 in the culture medium of lymphocytes.Results Liposome-entrapped gD,gD+HSP70,and HSP70-gD fusion protein all induced the production of IgG antibody against HSV-2gD,the proliferation of lymphocytes,and secretion of IFN-γand IL-4 by these lymphocytes.The serum level of IgG antibody against HSV-2gD,stimulation index of spleen lymphocytes,and levels of IFN-γand IL-4 were all significantly higher in mice immunized by liposome-entrapped HSP70-gD fusion protein than in those immunized by gD and gD+HSP70(all P<0.05).Conclusion These results suggest that HSP70 could promote the induction of specific immune response in mice by HSV-2gD.

Key words: Heat-shock proteins 70, Glycoproteins, Herpes simplex virus vaccine, Liposomes