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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+ breast cancer patients. PBMCs were tested for recognition of cyclin B1 peptides after one in vitro stimulation (A, B, E, and F) or no in vitro stimulation (C and D) in an IFN-γ ELISPOT assay. Number of T cells per well is indicated in parenthesis.
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fig2: HLA class I–restricted T cell responses to cyclin B1 peptides in HLA-A2+ breast cancer patients. PBMCs were tested for recognition of cyclin B1 peptides after one in vitro stimulation (A, B, E, and F) or no in vitro stimulation (C and D) in an IFN-γ ELISPOT assay. Number of T cells per well is indicated in parenthesis.

Mentions: We tested PBMCs from six breast cancer patients who had undergone surgery but had not yet started chemotherapy, for their ability to recognize the cyclin B1 peptides in an IFN-γ ELISPOT assay. Four out of the six HLA-A2+ breast cancer patients tested exhibited secondary responses against one or more of the cyclin B1 peptides (Fig. 2) . Patient A exhibited strong HLA class I–restricted secondary T cell responses to three of the three peptides tested, P4, CB9, and CB10, after only one in vitro stimulation. There was no recognition of the HIV-POL control peptide. Patient B appeared to have a weak secondary response to one of the three peptides tested, P1, and only after two in vitro stimulations. This patient was later found to be HLA-A*0206, suggesting that, if the anti-P1 response is real, P1 may also bind to HLA-A*0206.


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+ breast cancer patients. PBMCs were tested for recognition of cyclin B1 peptides after one in vitro stimulation (A, B, E, and F) or no in vitro stimulation (C and D) in an IFN-γ ELISPOT assay. Number of T cells per well is indicated in parenthesis.
© Copyright Policy
Related In: Results  -  Collection

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

fig2: HLA class I–restricted T cell responses to cyclin B1 peptides in HLA-A2+ breast cancer patients. PBMCs were tested for recognition of cyclin B1 peptides after one in vitro stimulation (A, B, E, and F) or no in vitro stimulation (C and D) in an IFN-γ ELISPOT assay. Number of T cells per well is indicated in parenthesis.
Mentions: We tested PBMCs from six breast cancer patients who had undergone surgery but had not yet started chemotherapy, for their ability to recognize the cyclin B1 peptides in an IFN-γ ELISPOT assay. Four out of the six HLA-A2+ breast cancer patients tested exhibited secondary responses against one or more of the cyclin B1 peptides (Fig. 2) . Patient A exhibited strong HLA class I–restricted secondary T cell responses to three of the three peptides tested, P4, CB9, and CB10, after only one in vitro stimulation. There was no recognition of the HIV-POL control peptide. Patient B appeared to have a weak secondary response to one of the three peptides tested, P1, and only after two in vitro stimulations. This patient was later found to be HLA-A*0206, suggesting that, if the anti-P1 response is real, P1 may also bind to HLA-A*0206.

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