A functionally significant allelic polymorphism in a T cell receptor V beta gene segment.
The effect of an allelic polymorphism in the BV1S1 gene segment on recognition of major histocompatibility complex (MHC)-peptide complexes by a specific T cell receptor (TCR) was studied using RBL 2H3 cells transfected with TCR-CD3 zeta chimeric receptors. An HLA-A2-restricted human immunodeficiency virus (HIV) pol-specific cytotoxic T lymphocyte (CTL) clone utilizing the BV1S1A2 gene in combination with AV2S1A2 was identified and the extracellular domains of the TCR were fused to CD3 zeta. In degranulation assays RBL 2H3 transfectants expressing this receptor maintained the specificity of the parental CTL clone. The allelic variant BV1S1A1N1 containing a glutamine for histidine substitution at position 48 in the loop of the second complementarity-determining region was generated by site-directed mutagenesis. Transfection of this molecule as a CD3 zeta chimera together with the original AV2S1A2 CD3 zeta molecule resulted in cell surface expression of both chains but a loss of recognition of HLA-A2 HIV pol peptide-pulsed targets. The effect of this polymorphism on MHC-peptide recognition supports current models of TCR MHC-peptide interaction and provides evidence for a functional role for polymorphism in the TCRV genes.
Vessey
S J
SJ
Institute of Molecular Medicine, John Radcliffe Hospital, Oxford, Great Britain.
Bell
J I
JI
Jakobsen
B K
BK
D000483:Alleles; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002999:Clone Cells; D003601:Cytotoxicity Tests, Immunologic; D015789:HLA-A2 Antigen / Q000235:genetics / Q000276:immunology / Q000502:physiology; D006801:Humans; D008969:Molecular Sequence Data; D011110:Polymorphism, Genetic / Q000276:immunology*; D051381:Rats; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000235:genetics* / Q000502:physiology*; D014407:Tumor Cells, Cultured
D015789:HLA-A2 Antigen; D016693:Receptors, Antigen, T-Cell, alpha-beta