Construction and Functional Test of HLA-A＊2402-Peptide Tetramer

HLA-A~*2402 is one of the most frequent HLA-A allele in Asian population.To construct HLA-A~*2402-peptidetetramers,the transmembrane and intracellular segments of HLA-A~*2402 cDNA were replaced with BSP sequenceto form a fusion gene of sHLA-A~*2402-BSP.The sHLA-A~*2402-BSP fusion protein and β2m were high-levelexpressed as insoluble aggregates in E.coli,and refoided to form an HLA-A~*2402-peptide monomeric complex bydilution method in the presence of an antigenic peptide.The HLA-A~*2402-peptide monomeric complex wasbiotinated and tetramized to prepare HLA-A~*2402-peptide tetramer.Then using the HLA-A~*2402-peptidetetramers to detect antigen-specific cytotoxic T lymphocyte(CTL)induced by artificial antigen presenting cell(aAPC)in vitro.The results showed that HLA-A~*2402-peptide tetramer was prepared correctly,and functional indetecting antigen-specific CTL in vitro,HLA-A~*2402-peptide monomeric and its multimeric complexes areexpected to provide a powerful tool for studying mechanisms of immune-related diseases in Asian populations.Cellular & Molecular Immunology.2005;2(2):145-149.

Construction and functional test of HLA-A*2402-peptide tetramer

HLA-A*2402 is one of the most frequent HLA-A allele in Asian population. To construct HLA-A*2402-peptide tetramers, the transmembrane and intracellular segments of HLA-A*2402 cDNA were replaced with BSP sequence to form a fusion gene of sHLA-A*2402-BSP. The sHLA-A*2402-BSP fusion protein and beta2m were high-level expressed as insoluble aggregates in E.coli, and refolded to form an HLA-A*2402-peptide monomeric complex by dilution method in the presence of an antigenic peptide. The HLA-A*2402-peptide monomeric complex was biotinated and tetramized to prepare HLA-A*2402-peptide tetramer. Then using the HLA-A*2402-peptide tetramers to detect antigen-specific cytotoxic T lymphocyte (CTL) induced by artificial antigen presenting cell (aAPC) in vitro. The results showed that HLA-A*2402-peptide tetramer was prepared correctly, and functional in detecting antigen-specific CTL in vitro, HLA-A*2402-peptide monomeric and its multimeric complexes are expected to provide a powerful tool for studying mechanisms of immune-related diseases in Asian populations.