Surface-epitope masking and expression cloning identifies the human prostate carcinoma tumor antigen gene PCTA-1 a member of the galectin gene family.
The selective production of monoclonal antibodies (mAbs) reacting with defined cell surface-expressed molecules is now readily accomplished with an immunological subtraction approach, surface-epitope masking (SEM). Using SEM, prostate carcinoma (Pro 1.5) mAbs have been developed that react with tumor-associated antigens expressed on human prostate cancer cell lines and patient-derived carcinomas. Screening a human LNCaP prostate cancer cDNA expression library with the Pro 1.5 mAb identifies a gene, prostate carcinoma tumor antigen-1 (PCTA-1). PCTA-1 encodes a secreted protein of approximately 35 kDa that shares approximately 40% sequence homology with the N-amino terminal region of members of the S-type galactose-binding lectin (galectin) gene family. Specific galectins are found on the surface of human and marine neoplastic cells and have been implicated in tumorigenesis and metastasis. Primer pairs within the 3' untranslated region of PCTA-1 and reverse transcription-PCR demonstrate selective expression of PCTA-1 by prostate carcinomas versus normal prostate and benign prostatic hypertrophy. These findings document the use of the SEM procedure for generating mAbs reacting with tumor-associated antigens expressed on human prostate cancers. The SEM-derived mAbs have been used for expression cloning the gene encoding this human tumor antigen. The approaches described in this paper, SEM combined with expression cloning, should prove of wide utility for developing immunological reagents specific for and identifying genes relevant to human cancer.
Su
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Department of Pathology, Columbia University, College of Physicians and Surgeons, New York, NY 10032, USA.
Lin
J
J
Shen
R
R
Fisher
P E
PE
Goldstein
N I
NI
Fisher
P B
PB
D000595:Amino Acid Sequence; D000818:Animals; D000951:Antigens, Neoplasm / Q000032:analysis / Q000096:biosynthesis*; D001483:Base Sequence; D002417:Cattle; D002460:Cell Line; D002645:Chickens; D003001:Cloning, Molecular; D017931:DNA Primers; D037161:Galectins; D006388:Hemagglutinins / Q000737:chemistry* / Q000235:genetics; D006801:Humans; D007150:Immunohistochemistry; D008297:Male; D051379:Mice; D008819:Mice, Nude; D008969:Molecular Sequence Data; D005810:Multigene Family; D009363:Neoplasm Proteins / Q000096:biosynthesis* / Q000737:chemistry* / Q000235:genetics; D016133:Polymerase Chain Reaction; D011467:Prostate / Q000166:cytology / Q000378:metabolism; D017430:Prostate-Specific Antigen / Q000032:analysis; D011471:Prostatic Neoplasms / Q000378:metabolism* / Q000473:pathology; D014176:Protein Biosynthesis; D051381:Rats; D017386:Sequence Homology, Amino Acid; D014162:Transfection; D014183:Transplantation, Heterologous; D014407:Tumor Cells, Cultured
D000951:Antigens, Neoplasm; D017931:DNA Primers; D037161:Galectins; D006388:Hemagglutinins; C091991:LGALS8 protein, human; D009363:Neoplasm Proteins; D017430:Prostate-Specific Antigen