Identification of the HERV-K gag antigen in prostate cancer by SEREX using autologous patient serum and its immunogenicity.
The prostate cancer HERV-K gag-related NGO-Pr-54 antigen was identified by SEREX analysis using autologous patient serum. NGO-Pr-54 mRNA was observed to be faintly expressed in normal prostate and strongly expressed in a variety of cancers, including ovarian cancer (5/8), prostate cancer (6/9), and leukemia (5/14). A phage plaque assay showed that a strong reaction was constantly observed with clone ZH042 in which the 5' end of NGO-Pr-54 is deleted, suggesting that it contained the sequence coding for the protein product. A TI-35 mAb was produced using a recombinant protein (438 aa) deduced from the sequence of ZH042. Transfection of clone ZH042 into 293T cells resulted in the production of an approximately 50-kDa molecule visualized by Western blotting. Natural production of the molecule was confirmed in a SK-MEL-23 melanoma cell line. An indirect immunofluorescence assay showed that NGO-Pr-54 protein was expressed on the cell surface as well as in the cytoplasm. Cell surface expression was confirmed by flow cytometry using the TI-35 mAb. The antibody response against NGO-Pr-54 was observed in patients with bladder (5.1%), liver (4.1%), lung (3.4%), ovarian (5.6%), and prostate (4.2%) cancer, as well as with malignant melanoma (13.2%).
Ishida
Toshiaki
T
Department of Immunology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, 2-5-1 Shikata-cho, Okayama 700-8558, Japan.
Obata
Yuichi
Y
Ohara
Nobuya
N
Matsushita
Hirokazu
H
Sato
Shuichiro
S
Uenaka
Akiko
A
Saika
Takashi
T
Miyamura
Takako
T
Chayama
Kosuke
K
Nakamura
Yurika
Y
Wada
Hisashi
H
Yamashita
Toshiharu
T
Morishima
Tsuneo
T
Old
Lloyd J
LJ
Nakayama
Eiichi
E
D000369:Aged, 80 and over; D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm / Q000235:genetics* / Q000276:immunology; D019556:COS Cells; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D020077:Endogenous Retroviruses; D005260:Female; D019084:Fluorescent Antibody Technique, Indirect; D015972:Gene Expression Regulation, Neoplastic; D015683:Gene Products, gag; D006801:Humans; D007106:Immune Sera / Q000032:analysis; D007938:Leukemia / Q000235:genetics / Q000276:immunology; D008297:Male; D008545:Melanoma / Q000235:genetics / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins / Q000235:genetics*; D010051:Ovarian Neoplasms / Q000235:genetics / Q000276:immunology; D011467:Prostate / Q000276:immunology / Q000378:metabolism; D011471:Prostatic Neoplasms / Q000235:genetics* / Q000276:immunology; D011994:Recombinant Proteins / Q000235:genetics; D014162:Transfection
D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm; D015683:Gene Products, gag; D007106:Immune Sera; C534523:NGO-Pr-54 protein, human; D009363:Neoplasm Proteins; D011994:Recombinant Proteins