Differential presentation of tumor antigen-derived epitopes by MHC-class I and antigen-positive tumor cells.
SSX2 is a member of the family of cancer/testis antigens. The SSX2 derived peptide SSX2(103-111) has been shown to be presented to cytotoxic T-lymphocytes (CTL) by Major-Histocompatibility (MHC) Class-I complexes after endogenous processing, more precisely by the allele HLA-A*0201. The HLA-A*0201- and SSX2-positive melanoma cell line SK-Mel-37 but not Me275 had been shown to elicit reactivity in SSX2(103-111) specific cytotoxic T-lymphocytes. To analyze the correlation between SSX2(103-111) presentation and T-cell stimulation, we intended to visualize presentation of SSX2(103-111) in these melanoma cell lines. Fab-antibodies were established from a human phage library with specificity for SSX2(103-111)/HLA-A*0201 complexes (but non-reactive with HLA-A*0201 or SSX2(103-111) alone) and used to visualize the presentation of SSX2(103-111) in the context of HLA-A*0201 by fluorescence microscopy. Presentation of SSX2(103-111) the context of HLA-A*0201 was demonstrated for the majority of SK-Mel-37, but for only a small fraction (<1%) of Me275 as indicated by a clear membrane-staining pattern in fluorescence microscopy. The presentation of SSX2(103-111) on SK-Mel37 and Me275, but not the expression of the SSX2 protein correlated with the capability of these cells to stimulate cells of an SSX2(103-111)-specific T-cell clone. MHC-peptide specific antibodies are a valuable tool for the analysis of antigenic peptides in the context of MHC-I molecules and for the structural definition of immunodominant epitopes.
Held
Gerhard
G
Jose-Carreras-Center for Immuno- and Genetherapy, Klinik für Innere Medizin I, Saarland University Medical School, 66421 Homburg, Saar, Germany. gerhard.held@uks.eu
Neumann
Frank
F
Sturm
Christine
C
Kaestner
Lars
L
Dauth
Nina
N
de Bruijn
Diederik R
DR
Renner
Christoph
C
Lipp
Peter
P
Pfreundschuh
Michael
M
D016475:3T3 Cells; D000818:Animals; D000918:Antibody Specificity; D017951:Antigen Presentation; D000951:Antigens, Neoplasm / Q000235:genetics / Q000276:immunology*; D001943:Breast Neoplasms / Q000235:genetics / Q000276:immunology*; D045744:Cell Line, Tumor; D015234:HLA-A Antigens / Q000276:immunology*; D015789:HLA-A2 Antigen; D006801:Humans; D007128:Immunoglobulin Fragments / Q000276:immunology; D008545:Melanoma / Q000235:genetics / Q000276:immunology*; D051379:Mice; D008856:Microscopy, Fluorescence; D009363:Neoplasm Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D010446:Peptide Fragments / Q000276:immunology; D016133:Polymerase Chain Reaction; D012097:Repressor Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology
D000951:Antigens, Neoplasm; D015234:HLA-A Antigens; C435939:HLA-A*02:01 antigen; D015789:HLA-A2 Antigen; D007128:Immunoglobulin Fragments; D009363:Neoplasm Proteins; D010446:Peptide Fragments; D012097:Repressor Proteins; C093962:synovial sarcoma X breakpoint proteins