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Identification of cyclin B1 as a shared human epithelial tumor-associated antigen recognized by T cells.

Kao H, Marto JA, Hoffmann TK, Shabanowitz J, Finkelstein SD, Whiteside TL, Hunt DF, Finn OJ - J. Exp. Med. (2001)

Bottom Line: We also detected memory T cells specific for one or more of these peptides in patients with breast cancer and squamous cell carcinomas of the head and neck (SCCHN).Immunohistochemical analysis of tumor lines and tissue sections showed cyclin B1 overexpression and aberrant localization in the cytoplasm instead of the nucleus.Sequencing genomic DNA and cDNA corresponding to P1-P6 region showed that differences in COOH-terminal residues were not due to either DNA mutations or errors in transcription, suggesting a high error rate in translation of cyclin B1 protein in tumors.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Genetics and Biochemistry, University of Pittsburgh, Pittsburgh, PA 15261, USA.

ABSTRACT
We eluted peptides from class I molecules of HLA-A2.1(+) breast adenocarcinoma and loaded reverse phase high-performance liquid chromatography (HPLC) fractions onto dendritic cells to prime naive CD8(+) T cells. Fractions that supported growth of tumor-specific cytotoxic T lymphocytes were analyzed by nano-HPLC micro-ESI tandem mass spectrometry. Six HLA-A2.1-binding peptides, four 9-mers (P1-P4) differing in the COOH-terminal residue, and two 10-mers (P5 and P6) with an additional COOH-terminal alanine, were identified in one fraction. Peptide sequences were homologous to cyclin B1. We primed CD8(+) T cells from another HLA-A2.1(+) healthy donor with synthetic peptides and generated P4-specific responses. We also detected memory T cells specific for one or more of these peptides in patients with breast cancer and squamous cell carcinomas of the head and neck (SCCHN). T cells from one patient, restimulated once in vitro, could kill the tumor cell line from which the peptides were derived. Immunohistochemical analysis of tumor lines and tissue sections showed cyclin B1 overexpression and aberrant localization in the cytoplasm instead of the nucleus. Sequencing genomic DNA and cDNA corresponding to P1-P6 region showed that differences in COOH-terminal residues were not due to either DNA mutations or errors in transcription, suggesting a high error rate in translation of cyclin B1 protein in tumors.

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HLA class I–restricted T cell responses to cyclin B1 peptides in HLA-A2.1+ SCCHN patients. PBMCs were tested for recognition of cyclin B1 peptides after one (A) or two in vitro stimulations (B–D) in an IFN-γ ELISPOT assay.
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fig3: HLA class I–restricted T cell responses to cyclin B1 peptides in HLA-A2.1+ SCCHN patients. PBMCs were tested for recognition of cyclin B1 peptides after one (A) or two in vitro stimulations (B–D) in an IFN-γ ELISPOT assay.

Mentions: We also examined PBMCs from five HLA-A*0201 SCCHN patients for their ability to respond to the cyclin B1 peptides in an IFN-γ ELISPOT assay. These patients were chosen for having either stable disease or being in long term remission. We detected HLA class I–restricted T cell responses to one or more of the cyclin B1 peptides in four of the five patients tested (Fig. 3) . Patient A exhibited HLA class I–restricted T cell responses to five of eight peptides, after only one in vitro stimulation. For two of these peptides, P4 and CB9, we also detected peptide-specific T cells in the absence of any in vitro stimulation (data not shown).


Identification of cyclin B1 as a shared human epithelial tumor-associated antigen recognized by T cells.

Kao H, Marto JA, Hoffmann TK, Shabanowitz J, Finkelstein SD, Whiteside TL, Hunt DF, Finn OJ - J. Exp. Med. (2001)

HLA class I–restricted T cell responses to cyclin B1 peptides in HLA-A2.1+ SCCHN patients. PBMCs were tested for recognition of cyclin B1 peptides after one (A) or two in vitro stimulations (B–D) in an IFN-γ ELISPOT assay.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2195974&req=5

fig3: HLA class I–restricted T cell responses to cyclin B1 peptides in HLA-A2.1+ SCCHN patients. PBMCs were tested for recognition of cyclin B1 peptides after one (A) or two in vitro stimulations (B–D) in an IFN-γ ELISPOT assay.
Mentions: We also examined PBMCs from five HLA-A*0201 SCCHN patients for their ability to respond to the cyclin B1 peptides in an IFN-γ ELISPOT assay. These patients were chosen for having either stable disease or being in long term remission. We detected HLA class I–restricted T cell responses to one or more of the cyclin B1 peptides in four of the five patients tested (Fig. 3) . Patient A exhibited HLA class I–restricted T cell responses to five of eight peptides, after only one in vitro stimulation. For two of these peptides, P4 and CB9, we also detected peptide-specific T cells in the absence of any in vitro stimulation (data not shown).

Bottom Line: We also detected memory T cells specific for one or more of these peptides in patients with breast cancer and squamous cell carcinomas of the head and neck (SCCHN).Immunohistochemical analysis of tumor lines and tissue sections showed cyclin B1 overexpression and aberrant localization in the cytoplasm instead of the nucleus.Sequencing genomic DNA and cDNA corresponding to P1-P6 region showed that differences in COOH-terminal residues were not due to either DNA mutations or errors in transcription, suggesting a high error rate in translation of cyclin B1 protein in tumors.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Genetics and Biochemistry, University of Pittsburgh, Pittsburgh, PA 15261, USA.

ABSTRACT
We eluted peptides from class I molecules of HLA-A2.1(+) breast adenocarcinoma and loaded reverse phase high-performance liquid chromatography (HPLC) fractions onto dendritic cells to prime naive CD8(+) T cells. Fractions that supported growth of tumor-specific cytotoxic T lymphocytes were analyzed by nano-HPLC micro-ESI tandem mass spectrometry. Six HLA-A2.1-binding peptides, four 9-mers (P1-P4) differing in the COOH-terminal residue, and two 10-mers (P5 and P6) with an additional COOH-terminal alanine, were identified in one fraction. Peptide sequences were homologous to cyclin B1. We primed CD8(+) T cells from another HLA-A2.1(+) healthy donor with synthetic peptides and generated P4-specific responses. We also detected memory T cells specific for one or more of these peptides in patients with breast cancer and squamous cell carcinomas of the head and neck (SCCHN). T cells from one patient, restimulated once in vitro, could kill the tumor cell line from which the peptides were derived. Immunohistochemical analysis of tumor lines and tissue sections showed cyclin B1 overexpression and aberrant localization in the cytoplasm instead of the nucleus. Sequencing genomic DNA and cDNA corresponding to P1-P6 region showed that differences in COOH-terminal residues were not due to either DNA mutations or errors in transcription, suggesting a high error rate in translation of cyclin B1 protein in tumors.

Show MeSH
Related in: MedlinePlus