A simple assay for detection of peptides promoting the assembly of HLA class I molecules.
Synthetic peptides derived from influenza virus and human immunodeficiency virus were tested for their ability to promote the assembly of HLA-A2 and HLA-B51 molecules in T2 cell lysates. Specific assembly was detected by an enzyme-linked immunosorbent assay. The most significant HLA-A2 assembly was obtained in the presence of peptides known to be targets for HLA-A2-restricted cytotoxic T lymphocytes (influenza matrix M.58-66 and HIV Pol 476-484). Three of a batch of Nef peptides corresponding to epitopic regions for cytotoxic T lymphocytes, caused significant assembly of HLA-A2 (Nef 83-91, 137-145 and 144-153), but only at high concentrations (100 microM). As these peptides bound relatively weakly, it is unlikely that they are good candidates for HLA-A2-restricted CTL epitopes. Peptides matrix M.60-68, Nef 186-194, and Plasmodium falciparum sh.77-85 produced the most significant assembly of HLA-B51. These peptides have a dominant hydrophobic anchor residue (V, L. I) at position 9 that could occupy pocket "F". Our results also suggest that another hydrophobic residue (V, L) at position 3 or 4 may anchor to hydrophobic pocket "D" of HLA-B51. Proline at position 2 greatly increases HLA-B51 anchoring.
Connan
F
F
Institut Cochin de Génétique Moléculaire (ICGM), INSERM U152, Paris, France.
Hlavac
F
F
Hoebeke
J
J
Guillet
J G
JG
Choppin
J
J
D000595:Amino Acid Sequence; D000818:Animals; D000953:Antigens, Protozoan / Q000737:chemistry / Q000378:metabolism*; D000956:Antigens, Viral / Q000737:chemistry / Q000378:metabolism*; D015693:Gene Products, nef / Q000737:chemistry / Q000378:metabolism; D015789:HLA-A2 Antigen / Q000378:metabolism*; D015235:HLA-B Antigens / Q000378:metabolism*; D059949:HLA-B51 Antigen; D009980:Influenza A virus / Q000276:immunology; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D010455:Peptides / Q000276:immunology* / Q000378:metabolism; D010963:Plasmodium falciparum / Q000276:immunology; D011485:Protein Binding; D013329:Structure-Activity Relationship
D000953:Antigens, Protozoan; D000956:Antigens, Viral; D015693:Gene Products, nef; D015789:HLA-A2 Antigen; D015235:HLA-B Antigens; D059949:HLA-B51 Antigen; D046911:Macromolecular Substances; D010455:Peptides