脂质体包裹HSV-2gD-HSP70融合蛋白诱导小鼠免疫应答的研究  

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

作　　者：樊建勇[1] 杨慧兰[1] 

机构地区：[1]广州军区广州总医院皮肤科,510010

出　　处：《中华皮肤科杂志》2007年第10期609-611,共3页Chinese Journal of Dermatology

基　　金：国家自然科学基金(30371290);广东省医学科研基金资助课题(A2006492)

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。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, lymphocytes were isolated from the spleen of the vaccinated mice and stimulated with HSV-2gD protein, the proliferation of lymphocytes was measured by MTT assay, and ELISA assay was utilized to measure the levels of IFN-γ and 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-γ and IL-4 by these lymphocytes. The serum level of IgG antibody against HSV-2gD, stimulation index of spleen lymphocytes, and levels of IFN-γ and 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.

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 

分 类 号：R392[医药卫生—免疫学]