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Identification of Breast Cancer-Restricted Antigens by Antibody Screening of SKBR3 cDNA Library Using a Preselected Patient's Serum

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Abstract

Screening of a breast cancer cDNA library from SKBR3 human breast cancer cells by SEREX (serological analysis of cDNA expression library) using a preselected serum from a breast cancer patient revealed 13 genes, two of which, INT-MI-1 and INT-MI-2, encode novel gene products, while the remaining 11 genes and their products are identical with or highly homologous to known GenBank entries. Immunoscreening of the 13 clones using 20 allogeneic sera from breast cancer patients and 20 samples from age- and gender-matched healthy donors showed that lactate dehydrogenase-A (LDH-A), lactate dehydrogenase-B (LDH-B), fibulin-1, and thyroid hormone-binding protein (THBP) were recognized principally by the breast cancer patient sera, indicating the immunogenicity of these molecules in vivo. The other antigens were similarly recognized by normal and patients sera, and thus not tumor-restricted immunologically. RT-PCR analysis revealed strong expression of fibulin-1 in tumor cell lines and surgical specimen whereas in the same experimental conditions, normal tissues scored negative. Also THBP expression was found in various tumors whereas in normal tissues, its expression is restricted to the testis and, at lower levels, in ovary, liver, and spleen. In contrast, LDH-A and LDH-B were ubiquitously expressed in normal and tumor tissues, with LDH-B levels considerably lower and heterogeneous in normal samples compared to those expressed in tumor cell lines. The differential expression of fibulin-1 between the normal tissues and breast carcinoma cell lines (5/6) and surgical specimens (5/6) suggests the possible involvement of the overexpression of this extracellular matrix-associated glycoprotein in the pathogenesis of this neoplasm.

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Authors and Affiliations

  1. Molecular Targeting Unit, Department of Experimental Oncology, USA

    Stefania Forti, AnnaMaria Invernizzi, Fabio Castiglioni, Serenella M. Pupa & Sylvie Ménard

  2. Ludwig Institute for Cancer Research, New York Branch at Memorial Sloan-Kettering Cancer Center, New York, NY, USA

    Matthew J. Scanlan & Lloyd J. Old

  3. Anesthesia and Intensive Care Unit, Italy

    Sandro Pupa

  4. General Oncology B Breast Unit, Italy

    Roberto Agresti

  5. Gynecological Oncology Unit, Italy

    Rosanna Fontanelli

  6. Medicine Laboratory Unit, Istituto Nazionale Tumori, Milano, Italy

    Daniele Morelli

Authors
  1. Stefania Forti
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  2. Matthew J. Scanlan
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  3. AnnaMaria Invernizzi
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Forti, S., Scanlan, M.J., Invernizzi, A. et al. Identification of Breast Cancer-Restricted Antigens by Antibody Screening of SKBR3 cDNA Library Using a Preselected Patient's Serum. Breast Cancer Res Treat 73, 245–256 (2002). https://doi.org/10.1023/A:1015854415746

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