Generation of heat shock protein-based vaccines by intracellular loading of gp96 with antigenic peptides
Generation of heat shock protein-based vaccines by intracellular loading of gp96 with antigenic peptides
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DOI：
10.1016/s0165-2478(97)00048-5
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发表时间：
1997-06-01
影响因子：
4.4
通讯作者：
Huckriede, A
中科院分区：
文献类型：
--
作者：
Heikema, A;Agsteribbe, E;Huckriede, A
几项研究表明，用从病毒感染细胞的肿瘤中纯化的愈合休克蛋白（HSPs）免疫诱导特异性细胞毒性T细胞（CTL）活性。这种免疫应答针对与HSP结合的肽，而不是针对HSP本身。所述肽衍生自肿瘤或病毒特异性蛋白质，所述蛋白质在正常蛋白质周转和加工过程中降解以通过MHC I类分子呈递。热休克蛋白似乎作为载体的抗原肽。在免疫后，它们确保它们被专门的巨噬细胞摄取，并将它们引入MHC I类呈递途径，否则该途径仅可用于细胞内蛋白质。使用流感病毒核蛋白（NP）作为模型抗原，我们已经测试了是否可以通过在纯化HSP之前在培养的细胞中过表达抗原来产生基于HSP的疫苗。采用基于Semliki Forest病毒（SFV）复制子的转染系统来实现NP的高表达。由于SFV介导的鼠细胞转染是低效的，我们使用仓鼠衍生的细胞系BHK 21作为负载NP肽的gp96的来源，其可以100%效率转染。从转染的细胞中纯化蛋白质并用于第一次接种研究。仓鼠gp96制剂在小鼠中耐受良好，未观察到针对外源蛋白的抗体应答。初步结果表明，确实诱导了针对NP的细胞免疫应答。SFV转染适用于任何已知的抗原，因此被认为是用于生产能够诱导细胞免疫应答的基于HSP的疫苗的优雅手段。(C)1997年Elsevier Science B.V.
Several studies have shown that immunization with heal shock proteins (HSPs) purified from tumors of virus-infected cells induces specific cytotoxic T-cell (CTL) activity. This immune response is directed against peptides bound to the HSPs rather than against the HSPs themselves. The peptides are derived from tumor- or virus-specific proteins which are degraded in the course of normal protein turnover and processing for presentation by MHC class I molecules. The HSPs appear to function as carriers for the antigenic peptides. Upon immunization they ensure their uptake by specialized macrophages and their introduction into the MHC class I presentation route which is otherwise accessible only for intracellular proteins. Using influenza virus nucleoprotein (NP) as a model antigen, we have tested whether an HSP-based vaccine can be produced by overexpressing an antigen in cultured cells prior to purification of the HSP's. The transfection system based on the Semliki Forest virus (SFV) replicon was employed to achieve high expression of NP. Since SFV-mediated transfection of murine cells was inefficient we used the hamster-derived cell line BHK21, which can be transfected with 100% efficiency, as a source for NP peptide-loaded gp96. The protein was purified from transfected cells and used for first vaccination studies. The hamster gp96 preparation was well tolerated in mice, an antibody response against the foreign protein was not observed. Preliminary results suggest that a cellular immune response against NP was indeed induced. SFV transfection is applicable for any known antigen and is therefore considered to be an elegant means for the production of HSP-based vaccines capable of inducing a cellular immune response. (C) 1997 Elsevier Science B.V.