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.
Lu
Xiaoling
X
Department of Immunology, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.
Wu
Xiongwen
X
Liang
Zhihui
Z
Weng
Xiufang
X
Li
Qing
Q
Gong
Feili
F
D000483:Alleles; D000595:Amino Acid Sequence; D000938:Antigen-Presenting Cells / Q000276:immunology; D001208:Asia; D001483:Base Sequence; D015234:HLA-A Antigens / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D066298:In Vitro Techniques; D046912:Multiprotein Complexes; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology
D015234:HLA-A Antigens; D046912:Multiprotein Complexes; D011993:Recombinant Fusion Proteins