Li Pira Giuseppina, Ivaldi Federico, Moretti Paolo, Manca Fabrizio
Laboratory of Cellular Immunology, Advanced Biotechnology Center, 16132 Genoa, Italy.
J Biomed Biotechnol. 2010;2010:325720. doi: 10.1155/2010/325720. Epub 2010 Jun 15.
Mapping of antigenic peptide sequences from proteins of relevant pathogens recognized by T helper (Th) and by cytolytic T lymphocytes (CTL) is crucial for vaccine development. In fact, mapping of T-cell epitopes provides useful information for the design of peptide-based vaccines and of peptide libraries to monitor specific cellular immunity in protected individuals, patients and vaccinees. Nevertheless, epitope mapping is a challenging task. In fact, large panels of overlapping peptides need to be tested with lymphocytes to identify the sequences that induce a T-cell response. Since numerous peptide panels from antigenic proteins are to be screened, lymphocytes available from human subjects are a limiting factor. To overcome this limitation, high throughput (HTP) approaches based on miniaturization and automation of T-cell assays are needed. Here we consider the most recent applications of the HTP approach to T epitope mapping. The alternative or complementary use of in silico prediction and experimental epitope definition is discussed in the context of the recent literature. The currently used methods are described with special reference to the possibility of applying the HTP concept to make epitope mapping an easier procedure in terms of time, workload, reagents, cells and overall cost.
绘制辅助性T细胞(Th)和细胞毒性T淋巴细胞(CTL)识别的相关病原体蛋白质的抗原肽序列图谱,对于疫苗开发至关重要。事实上,T细胞表位图谱分析为基于肽的疫苗设计以及用于监测受保护个体、患者和疫苗接种者中特异性细胞免疫的肽库设计提供了有用信息。然而,表位图谱分析是一项具有挑战性的任务。实际上,需要用淋巴细胞测试大量重叠肽,以鉴定诱导T细胞应答的序列。由于要筛选来自抗原蛋白的众多肽组,人类受试者可得的淋巴细胞是一个限制因素。为克服这一限制,需要基于T细胞检测小型化和自动化的高通量(HTP)方法。本文我们考虑HTP方法在T表位图谱分析中的最新应用。在近期文献的背景下,讨论了计算机预测和实验性表位定义的替代或互补使用。特别参考将HTP概念应用于使表位图谱分析在时间、工作量、试剂、细胞和总体成本方面成为更简便程序的可能性,描述了当前使用的方法。