Flow cytometric analysis of peptide binding to major histocampatibility complex class I for hepatitis C virus core T‐cell epitopes

• Published in: Cytometry (New York, N.Y.) N.Y.), 2000-12, Vol.41 (4), p.271-278
• Main Authors: Schweitzer, Susann, Schneiders, Angelika M., Langhans, Bettina, Kraas, Wolfgang, Jung, Günther, Vidalin, Olivier, Inchauspe, Genevieve, Sauerbruch, Tilman, Spengler, Ulrich
• Format: Article
• Language: English
• Subjects: Animals; B-Lymphocytes - immunology; Binding, Competitive; Biotinylation; Cells, Cultured; Epitope Mapping - methods; Epitopes, T-Lymphocyte - immunology; Epitopes, T-Lymphocyte - metabolism; Flow Cytometry - methods; Fluorescent Dyes; hepatitis C virus; Histocompatibility Antigens Class I - immunology; Histocompatibility Antigens Class I - metabolism; Humans; major histocompatibility complex class I; Mice; Mice, Inbred BALB C; Oligopeptides - immunology; Oligopeptides - metabolism; Protein Binding; Spleen - immunology; T-Lymphocytes, Cytotoxic - immunology; T-Lymphocytes, Cytotoxic - metabolism; T‐cell epitopes; Viral Core Proteins - immunology; Viral Core Proteins - metabolism
• ISSN: 0196-4763; 1097-0320
• DOI: 10.1002/1097-0320(20001201)41:4<271::AID-CYTO5>3.0.CO;2-M

Background/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. Cytometry 41:271–278, 2000. © 2000 Wiley‐Liss, Inc.