Limits...
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.

Show MeSH

Related in: MedlinePlus

T cells from an HLA-A2.1+ SCCHN patient restimulated to cyclin B1 peptides in vitro are able to kill the original tumor. T cells were restimulated with P4 (A), and CB9 (B) for 5 d and tested in a CTL assay.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC2195974&req=5

fig4: T cells from an HLA-A2.1+ SCCHN patient restimulated to cyclin B1 peptides in vitro are able to kill the original tumor. T cells were restimulated with P4 (A), and CB9 (B) for 5 d and tested in a CTL assay.

Mentions: Because T cells from healthy individuals primed to synthetic cyclin B1 peptides in vitro could not kill tumor cells, we wanted to determine whether T cells from patients that we assumed had been primed in vivo could lyse tumor cells. We restimulated PBLs from one SCCHN patient once in vitro with either peptide P4 or peptide CB9 and tested the T cells for their ability to kill the original tumor MS-A2 from which the peptides were derived. We included in the assay the same tumor transduced with the CD80 gene to provide “costimulation” for T cell activation (MS-A2/CD80). As shown in Fig. 4 A, T cells restimulated once with P4 were able to lyse the MS-A2/CD80 tumor and to a lesser extent, MS-A2 tumor, but not the HLA-A2− target MS or K562 that was a control target for LAK activity. Similarly, in Fig. 4 B, T cells restimulated with CB9 peptide were able to lyse the MS-A2+/CD80+ tumor and not the other targets.


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)

T cells from an HLA-A2.1+ SCCHN patient restimulated to cyclin B1 peptides in vitro are able to kill the original tumor. T cells were restimulated with P4 (A), and CB9 (B) for 5 d and tested in a CTL assay.
© Copyright Policy
Related In: Results  -  Collection

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

fig4: T cells from an HLA-A2.1+ SCCHN patient restimulated to cyclin B1 peptides in vitro are able to kill the original tumor. T cells were restimulated with P4 (A), and CB9 (B) for 5 d and tested in a CTL assay.
Mentions: Because T cells from healthy individuals primed to synthetic cyclin B1 peptides in vitro could not kill tumor cells, we wanted to determine whether T cells from patients that we assumed had been primed in vivo could lyse tumor cells. We restimulated PBLs from one SCCHN patient once in vitro with either peptide P4 or peptide CB9 and tested the T cells for their ability to kill the original tumor MS-A2 from which the peptides were derived. We included in the assay the same tumor transduced with the CD80 gene to provide “costimulation” for T cell activation (MS-A2/CD80). As shown in Fig. 4 A, T cells restimulated once with P4 were able to lyse the MS-A2/CD80 tumor and to a lesser extent, MS-A2 tumor, but not the HLA-A2− target MS or K562 that was a control target for LAK activity. Similarly, in Fig. 4 B, T cells restimulated with CB9 peptide were able to lyse the MS-A2+/CD80+ tumor and not the other targets.

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