Flow cytometric analysis of peptide binding to major histocampatibility complex class I for hepatitis C virus core T-cell epitopes.
METHODS
To characterize the repertoire of T-cell epitopes on the hepatitis C virus (HCV) core protein, we studied major histocompatibility complex (MHC) class I binding of 75 decapeptides on 20 human B-cell lines and murine spleen cells using a flow cytometric assay. The results were compared with MHC class I stabilization on T2 cells, the SYFPEITHI algorithm, and known T-cell epitopes from the literature.


RESULTS
Binding of peptides proved to be specific for MHC class I molecules. We observed peak fluorescence signals at positions amino acids (aa) 35-44, aa 87-96, aa 131-140, and aa 167-176 in virtually all HLA-A2-positive cell lines. These sites corresponded to T-cell epitopes predicted by SYFPEITHI and the positions of known T-cell epitopes, whereas T2 stabilization was at variance for two peptides. The assay was applied to HLA-A2-negative cells and murine spleen cells without further modification, and identified additional peptides, corresponding to known T-cell epitopes.


CONCLUSIONS
Peptide binding to different MHC class I alleles can be mapped rapidly by a flow cytometric assay and enables a first orientation on the sites of possible T-cell epitopes. Application of this assay to HCV core suggests a rather limited repertoire of epitopes in the Caucasoid population.
Schweitzer
S
S
Department of Internal Medicine I, University of Bonn, Bonn, Germany.
Schneiders
A M
AM
Langhans
B
B
Kraas
W
W
Jung
G
G
Vidalin
O
O
Inchauspe
G
G
Sauerbruch
T
T
Spengler
U
U
D000818:Animals; D001402:B-Lymphocytes / Q000276:immunology; D001667:Binding, Competitive; D019921:Biotinylation; D002478:Cells, Cultured; D018604:Epitope Mapping / Q000379:methods*; D018984:Epitopes, T-Lymphocyte / Q000276:immunology / Q000378:metabolism*; D005434:Flow Cytometry / Q000379:methods*; D005456:Fluorescent Dyes; D015395:Histocompatibility Antigens Class I / Q000276:immunology / Q000378:metabolism*; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D009842:Oligopeptides / Q000276:immunology / Q000378:metabolism; D011485:Protein Binding; D013154:Spleen / Q000276:immunology; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology / Q000378:metabolism; D014758:Viral Core Proteins / Q000276:immunology* / Q000378:metabolism*
D018984:Epitopes, T-Lymphocyte; D005456:Fluorescent Dyes; D015395:Histocompatibility Antigens Class I; D009842:Oligopeptides; D014758:Viral Core Proteins; C069130:nucleocapsid protein, Hepatitis C virus