Identification of peptide sequences that potentially trigger HLA-A2.1-restricted cytotoxic T lymphocytes.
We used the human processing defective cell line 174CEM.T2 (T2) to identify potential cytotoxic T lymphocyte (CTL) epitopes of human proteins. Exogenously added peptides can increase the number of properly folded HLA-A2.1 molecules on the cell surface of T2 cells, as shown by immunofluorescence measurements using the mouse monoclonal antibody BB7.2 (anti-HLA-A2.1) and fluorescein isothiocyanate-labeled goat anti-mouse F(ab')2 antibody. The peptides were selected on the basis of a computer score derived from the recently described HLA-A2.1 specific motif. Analysis of the influenza matrix protein showed that 15 out of 35 high-scoring peptides up-regulate the expression of HLA-A2.1 molecules on the T2 cell surface. The combination of the computer scoring program and an immunofluorescence-based peptide binding assay allows rapid detection of potential CTL target peptides.
Nijman
H W
HW
Department of Immunohematology and Blood Bank, University Hospital, Leiden, The Netherlands.
Houbiers
J G
JG
Vierboom
M P
MP
van der Burg
S H
SH
Drijfhout
J W
JW
D'Amaro
J
J
Kenemans
P
P
Melief
C J
CJ
Kast
W M
WM
D000595:Amino Acid Sequence; D000956:Antigens, Viral / Q000276:immunology*; D003602:Cytotoxicity, Immunologic; D000939:Epitopes; D015789:HLA-A2 Antigen / Q000276:immunology*; D006801:Humans; D007111:Immunity, Cellular; D066298:In Vitro Techniques; D009980:Influenza A virus / Q000276:immunology; D008969:Molecular Sequence Data; D010455:Peptides / Q000276:immunology*; D013329:Structure-Activity Relationship; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D014763:Viral Matrix Proteins / Q000737:chemistry / Q000276:immunology*; D001613:beta 2-Microglobulin / Q000276:immunology
D000956:Antigens, Viral; D000939:Epitopes; D015789:HLA-A2 Antigen; C031540:M-protein, influenza virus; D010455:Peptides; D014763:Viral Matrix Proteins; D001613:beta 2-Microglobulin