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Molecular profiling of the immune response in colon cancer using protein microarrays: Occurrence of autoantibodies to ubiquitin C ‐terminal hydrolase L3
Author(s) -
Nam Myeong J.,
MadozGurpide Juan,
Wang Hong,
Lescure Pascal,
Schmalbach Cecelia E.,
Zhao Rong,
Misek David E.,
Kuick Rork,
Brenner Dean E.,
Hanash Samir M.
Publication year - 2003
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200300594
Subject(s) - colorectal cancer , autoantibody , protein microarray , lung cancer , immune system , cancer , antibody microarray , antibody , antigen , proteomics , blot , biology , microbiology and biotechnology , cancer research , microarray , immunology , medicine , pathology , biochemistry , gene expression , gene
We implemented a protein microarray approach to identify proteins that induce a humoral response in colon cancer. Solubilized proteins from the LoVo colon adenocarcinoma cell line were separated into 1760 fractions, arrayed onto nitrocellulose‐coated slides, and hybridized with individual sera from 15 newly diagnosed patients with colon cancer, 15 with lung cancer, and 15 healthy subjects. 39/1760 fractions showed enhanced reactivity with sera from patients with colon cancer ( p < 0.01) relative to healthy controls. A distinct pattern of reactivity was observed with sera from colon cancer relative to lung cancer. One fraction that exhibited reactivity with 9/15 colon cancer sera was subjected to mass spectrometry leading to the identification of ubiquitin C ‐terminal hydrolase isozyme 3 (UCH‐L3) as a constituent. To validate the occurrence of autoantibodies to UCH‐L3, independent analysis was done by means of Western blots. UCH‐L3 antibodies were detected in 19/43 sera from patients with colon cancer, and in 0/54 sera from subjects with lung cancer (24), colon adenoma (15) or otherwise healthy (15). Our findings indicate the occurrence of an immune response to a broad set of antigens in colon cancer and the feasibility of identifying the antigenic targets using a combination of protein microarrays and mass spectrometry.