Identification of HLA class I-restricted tumor-associated antigens in adult T cell leukemia cells by mass spectrometric analysis.
OBJECTIVE
In this study, we attempted a comprehensive analysis of MHC class I-bound peptides in adult T cell leukemia (ATL) cells in order to identify as many tumor-associated antigens (TAAs) as possible that could be used for CTL-based immunotherapy.


RESULTS
Using mass spectrometry combined with reversed-phase liquid chromatography, we could sequence 188 HLA class I-restricted candidate peptides from three ATL-derived cell lines. In accordance with the restrained expression of HTLV-I viral RNA in these cell lines, there were no HTLV-I-encoded peptides among these candidates. Based on the differential expression between ATL cells and normal CD4+ T cells, we selected 10 novel peptides as T cell epitopes of overexpressed source proteins. RT-PCR analysis revealed that 5 source proteins including PRAME, a known tumor-testis antigen, were highly expressed in the majority of 16 ATL cases. Furthermore we could induce PRAME-specific CTLs in vitro from an HLA-B62+ healthy donor that showed specific cytotoxicity against HLA-B62+ PRAME+ ATL cells.


CONCLUSIONS
These results demonstrate that comprehensive analysis of HLA class I-bound peptides by mass spectrometry is useful for identification of TAA-derived peptides in ATL. Considering that expression patterns of leukemia/lymphoma-associated antigens vary from case to case, this approach appears to be suitable for the tailor-made immunotherapy of hematological malignancies.
Kawahara
Masahiro
M
Department of Hematology and Oncology, Graduate School of Medicine, Kyoto University, Kyoto, Japan.
Hori
Toshiyuki
T
Matsubara
Yasushi
Y
Okawa
Katsuya
K
Uchiyama
Takashi
T
D000951:Antigens, Neoplasm / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D045744:Cell Line, Tumor; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D002853:Chromatography, Liquid / Q000379:methods; D003601:Cytotoxicity Tests, Immunologic; D015395:Histocompatibility Antigens Class I / Q000096:biosynthesis* / Q000737:chemistry; D015368:Human T-lymphotropic virus 1 / Q000235:genetics; D006801:Humans; D016130:Immunophenotyping; D015459:Leukemia-Lymphoma, Adult T-Cell / Q000235:genetics / Q000276:immunology* / Q000821:virology; D013058:Mass Spectrometry / Q000379:methods; D008869:Microtubule-Associated Proteins / Q000096:biosynthesis / Q000235:genetics; D010446:Peptide Fragments / Q000032:analysis*; D017346:Protein Serine-Threonine Kinases / Q000096:biosynthesis / Q000235:genetics; D011506:Proteins / Q000235:genetics / Q000378:metabolism; D012367:RNA, Viral / Q000096:biosynthesis; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods
D000951:Antigens, Neoplasm; C454180:CKAP5 protein, human; D015395:Histocompatibility Antigens Class I; D008869:Microtubule-Associated Proteins; C404944:PRAME protein, human; C524815:PRR11 protein, human; D010446:Peptide Fragments; D011506:Proteins; D012367:RNA, Viral; D017346:Protein Serine-Threonine Kinases; C114482:CDKL5 protein, human