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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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Cyclin B1 protein is overexpressed in epithelial tumor cells. (A) MS-A2 cells, original magnification, ×20. (B) MS-A2 cells, original magnification, ×40. (C) 201T cells, original magnification, ×20. (D) 201T cells, original magnification, ×40. (E) Human airway bronchoepithelial cells, original magnification, ×20.
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fig5: Cyclin B1 protein is overexpressed in epithelial tumor cells. (A) MS-A2 cells, original magnification, ×20. (B) MS-A2 cells, original magnification, ×40. (C) 201T cells, original magnification, ×20. (D) 201T cells, original magnification, ×40. (E) Human airway bronchoepithelial cells, original magnification, ×20.

Mentions: To understand the reason why cyclin B1 peptides would elicit T cell responses in cancer patients, we examined by immunohistochemistry the cyclin B1 expression in the original tumor cell line MS-A2 from where they were first isolated and identified (Fig. 5, A and B) . There was intense staining of cyclin B1 protein in the tumor cells, predominantly found in the cytoplasm. Fig. 5, C and D, depict similar intense cytoplasmic staining of cyclin B1 in a human lung adenocarcinoma cell line 201T. No cyclin B1 staining was observed in normal cells, represented by primary cultures of human airway bronchoepithelial cells (Fig. 5 E).


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)

Cyclin B1 protein is overexpressed in epithelial tumor cells. (A) MS-A2 cells, original magnification, ×20. (B) MS-A2 cells, original magnification, ×40. (C) 201T cells, original magnification, ×20. (D) 201T cells, original magnification, ×40. (E) Human airway bronchoepithelial cells, original magnification, ×20.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2195974&req=5

fig5: Cyclin B1 protein is overexpressed in epithelial tumor cells. (A) MS-A2 cells, original magnification, ×20. (B) MS-A2 cells, original magnification, ×40. (C) 201T cells, original magnification, ×20. (D) 201T cells, original magnification, ×40. (E) Human airway bronchoepithelial cells, original magnification, ×20.
Mentions: To understand the reason why cyclin B1 peptides would elicit T cell responses in cancer patients, we examined by immunohistochemistry the cyclin B1 expression in the original tumor cell line MS-A2 from where they were first isolated and identified (Fig. 5, A and B) . There was intense staining of cyclin B1 protein in the tumor cells, predominantly found in the cytoplasm. Fig. 5, C and D, depict similar intense cytoplasmic staining of cyclin B1 in a human lung adenocarcinoma cell line 201T. No cyclin B1 staining was observed in normal cells, represented by primary cultures of human airway bronchoepithelial cells (Fig. 5 E).

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