[Epitope of somatic mutations in the hepatocyte growth factor receptor naturally processed and presented by HLA-A2 on human hepatocellular carcinoma].
OBJECTIVE
To isolate and identify peptides bound to HLA I on human hepatocellular carcinoma.


METHODS
MHC-associated peptides were extracted by mild acid wash of viable hepatocellular carcinomas cells, collected by gel filtration, and fractioned by reversed phase high pressured liquid chromatography (RP-HPLC). Peptides of individual fractions were reconstitution of T cell epitopes and were identified by cytotoxicity T lymophacyte assay. Actively extracted sample analyses were performed by HPLC-MS-MS (tandom mass spectrometry). Protein database in the internet was used as an additional tool for structure analysis and for determination of protein source of the eluted peptides.


RESULTS
RP-HPLC showed that there were over 20 different fractions of peptides derived from human hepatocellular carcinoma, and only two active peaks were identified by cytotoxicity T lymophocyte assay. The most promising candidate for T cell epitope was nonamers peptide (SLIVHLNEV), derived from met/hepatocyte growth factor receptor, point mutation was 1180F to L.


CONCLUSIONS
Sensitive sequencing by HPLC-MS may provide a powerful method of identifying tumor specific antigenic peptides, and nonamers peptide (SLIVHLNEV) is the hepatocellular carcinoma antigenic peptide.
Guo
A
A
Department of Pathology, Fourth Military Medical University, Xi'an 710032, China.
Sui
Y
Y
Ye
J
J
D000595:Amino Acid Sequence; D006528:Carcinoma, Hepatocellular / Q000235:genetics / Q000276:immunology* / Q000473:pathology; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003601:Cytotoxicity Tests, Immunologic; D018984:Epitopes, T-Lymphocyte / Q000032:analysis* / Q000235:genetics / Q000276:immunology; D015789:HLA-A2 Antigen / Q000276:immunology*; D006801:Humans; D020014:K562 Cells; D008113:Liver Neoplasms / Q000235:genetics / Q000276:immunology / Q000473:pathology; D009154:Mutation; D019859:Proto-Oncogene Proteins c-met / Q000235:genetics* / Q000276:immunology
D018984:Epitopes, T-Lymphocyte; D015789:HLA-A2 Antigen; D019859:Proto-Oncogene Proteins c-met