Simultaneous analysis of HER-2/neu gene amplification and protein overexpression in single cells of pleural and ascitic effusions from patients with breast and ovarian cancer

Vaccination of patients with cancer using dendritic cells (DCs) was shown to be effective for B-cell lymphoma and malignant melanoma. Here we provide evidence that patients with advanced breast and ovarian cancer can be efficiently vaccinated with autologous DCs pulsed with HER-2/neu– or MUC1-derived peptides. Ten patients were included in this pilot study. The DC vaccinations were well tolerated with no side effects. In 5 of 10 patients, peptide-specific cytotoxic T lymphocytes (CTLs) could be detected in the peripheral blood using both intracellular IFN-γ staining and 51Cr-release assays. The major CTL response in vivo was induced with the HER-2/neu–derived E75 and the MUC1-derived M1.2 peptide, which lasted for more than 6 months, suggesting that these peptides might be immunodominant. In addition, in one patient vaccinated with the MUC1-derived peptides, CEA- and MAGE-3 peptide-specific T-cell responses were detected after several vaccinations. In a second patient immunized with the HER-2/neu peptides, MUC1-specific T lymphocytes were induced after 7 immunizations, suggesting that antigen spreading in vivo might occur after successful immunization with a single tumor antigen. Our results show that vaccination of DCs pulsed with a single tumor antigen may induce immunologic responses in patients with breast and ovarian cancer. This study may be relevant to the design of future clinical trials of other peptide-based vaccines.

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Studies of the HER-2/neu proto-oncogene in human breast and ovarian cancer

Science ◽

10.1126/science.2470152 ◽

1989 ◽

Vol 244 (4905) ◽

pp. 707-712 ◽

Cited By ~ 4538

Author(s):

D. Slamon ◽

W Godolphin ◽

L. Jones ◽

J. Holt ◽

S. Wong ◽

...

Keyword(s):

Ovarian Cancer ◽

Human Breast ◽

Breast And Ovarian Cancer ◽

Her 2

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No Strong Association Between HER-2/neu Protein Overexpression and Gene Amplification in High-grade Invasive Urothelial Carcinomas

Pathology & Oncology Research ◽

10.1007/s12253-008-9027-y ◽

2008 ◽

Vol 14 (3) ◽

pp. 261-266 ◽

Cited By ~ 39

Author(s):

Vildan Caner ◽

Nilay Sen Turk ◽

Fusun Duzcan ◽

N. Lale Satiroglu Tufan ◽

E. Canan Kelten ◽

...

Keyword(s):

Gene Amplification ◽

Strong Association ◽

High Grade ◽

Protein Overexpression ◽

Urothelial Carcinomas ◽

Her 2

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HER-2/neu Amplification Testing in Breast Cancer by Multiplex Ligation-Dependent Probe Amplification in Comparison with Immunohistochemistry and In Situ Hybridization

Analytical Cellular Pathology ◽

10.1155/2009/615178 ◽

2009 ◽

Vol 31 (1) ◽

pp. 1-10

Author(s):

Cathy B. Moelans ◽

Roel A. de Weger ◽

Marja T.M. van Blokland ◽

Chantal Ezendam ◽

Sabrina Elshof ◽

...

Keyword(s):

Breast Cancer ◽

Gene Amplification ◽

Invasive Breast Cancer ◽

Protein Overexpression ◽

Quick Method ◽

Cheap Method ◽

High Concordance ◽

Her 2 ◽

Dependent Probe

Background: Assessment of HER-2/neu status in invasive breast cancer is crucial to establish eligibility for trastuzumab and taxane based chemotherapy. Next to immunohistochemistry (IHC) to evaluate protein overexpression, a second line gene amplification test is required for cases with equivocal protein expression. This study aimed to validate a new PCR based test, called Multiplex Ligation-dependent Probe Amplification (MLPA), as a simple and quick method to assess HER-2/neu gene amplification status in invasive breast cancer.Methods: MPLA results were compared with gene amplification status assessed by fluorescence in situ hybridization (FISH) and chromogenic in situ hybridization (CISH) as gold standard, and with protein overexpression by IHC in 518 breast carcinoma patients.Results: About 10% of cases overexpressed HER-2/neu at the protein level (IHC), and 11% of cases showed gene-amplification by MLPA. A high concordance was found between FISH and CISH, MLPA and IHC, and MLPA and CISH. MLPA showed amplification in 7/36 (19%) of the equivocal IHC 2+ cases. However, of the IHC 0/1+ cases, 6/434 (1.4%) were also amplified by MLPA, and amplification was confirmed in all of these cases by FISH/CISH. On the other hand, one of the 48 (2%) IHC 3+ cases was normal by MLPA and lack of amplification was confirmed by FISH/CISH.Conclusion: MLPA is a fast, accurate and cheap method to detect breast cancer HER-2/neu amplification in small quantities of DNA extracted from paraffin blocks, and thereby a reliable alternative to FISH and CISH.

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