An optimal viral peptide recognized by CD8+ T cells binds very tightly to the restricting class I major histocompatibility complex protein on intact cells but not to the purified class I protein.
CD8+ cytotoxic T lymphocytes recognize cell surface complexes formed by class I major histocompatibility complex (MHC-I) glycoproteins and antigenic peptides. We have identified a peptide nonamer (termed IV9) derived from the human immunodeficiency virus that is over a millionfold more active (at subpicomolar concentrations) than peptide analogues longer or shorter by one or two amino acid residues. Although IV9 does not detectably bind to isolated MHC-I molecules as measured by equilibrium dialysis, we quantitated its specific binding in unaltered form to MHC-I on intact cells. Less than 1% of cell surface MHC-I forms complexes with IV9, which suffices to trigger maximal cytotoxic T-lymphocyte activity. By contrast, a peptide dodecamer that includes the IV9 sequence and is active at micromolar concentrations does not bind to MHC-I on intact cells, raising the possibility that this longer peptide undergoes processing. Using stoichiometrically iodinated IV9 to obviate the ambiguities associated with trace labeling methods, we measured the dissociation kinetics of purified peptide/MHC-I complexes isolated by affinity chromatography and found these complexes to be exceedingly stable (t1/2 = 200-600 hr).
Tsomides
T J
TJ
Department of Biology, Massachusetts Institute of Technology, Cambridge 02139.
Walker
B D
BD
Eisen
H N
HN
D000595:Amino Acid Sequence; D016827:CD8 Antigens / Q000276:immunology*; D002460:Cell Line; D003602:Cytotoxicity, Immunologic; D006678:HIV / Q000201:enzymology / Q000276:immunology*; D015395:Histocompatibility Antigens Class I / Q000276:immunology* / Q000302:isolation & purification; D006801:Humans; D007700:Kinetics; D008969:Molecular Sequence Data; D010455:Peptides / Q000138:chemical synthesis / Q000276:immunology*; D011485:Protein Binding; D012194:RNA-Directed DNA Polymerase / Q000276:immunology*; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D014764:Viral Proteins / Q000276:immunology*
D016827:CD8 Antigens; D015395:Histocompatibility Antigens Class I; D010455:Peptides; D014764:Viral Proteins; D012194:RNA-Directed DNA Polymerase