Cytokeratin 8 and 19 as antigens recognized by adenocarcinoma-reactive human monoclonal antibody AE6F4

Akira Ichikawa, Hirofumi Tachibana, Seiji Kawamoto, Masanori Kamei, Tsutomu Honjoh, Shuichi Hashizume, Sanetaka Shirahata

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Abstract

The human monoclonal antibody (MAb) AE6F4 is secreted by a human-human hybridoma line established from the in vitro immunization of normal human peripheral blood lymphocytes with the human lung adenocarcinomn cell line, A549. This MAb is strongly reactive to lung cancer tissues. In the previous study, the antigens recognized by the MAb AE6F4 were purified from A549 cells and identified as 14-3-3 protein and 31kDa cytosolic phospholipase A2 (cPLA2). The MAb AE6F4 also binds two kinds of antigens (53kDA and 40kDa), which are not related to 14-3-3 protein or 31 kDa cPLA2, in the human breast adenocarcinoma cell line, MCF-7. We purified a 38 kDa antigen, which is a degradation product of 53kDa antigen from breast adenocarcinoma MCF-7 cells using ion-exchange and hydroxyapatite column chromatography. Two partial amino acid sequences of the purified 38 kDa antigen showed 95-100% homology to human cytokeratin 8 (CK8). Two-dimensional gel electrophoresis and immunoblot analysis of intermediate filament fraction separated from MCF-7 cells demonstrated that the 53kDa and 40kDa antigens were CK8 and CK19, respectively. Antigenic determinants on CK8 and CK19 recognized by the MAb AE6F4 were resistant to sodium periodate treatment, although antigenic determinant on 31 kDa antigen (14-3-3 protein and(or) cPLA2) was sensitive to this treatment. These results suggest that the MAb AE6F4 reacts with both carbohydrate and peptide antigenic determinants.

Original languageEnglish
Pages (from-to)195-202
Number of pages8
JournalHuman Antibodies
Volume8
Issue number4
Publication statusPublished - Dec 4 1997

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Keratin-8
Keratin-19
Adenocarcinoma
Monoclonal Antibodies
Antigens
Cytosolic Phospholipases A2
14-3-3 Proteins
Epitopes
MCF-7 Cells
Breast
Intermediate Filaments
Ion Exchange
Electrophoresis, Gel, Two-Dimensional
Hybridomas
Durapatite
Chromatography
Amino Acid Sequence
Immunization
Lung Neoplasms
Carbohydrates

All Science Journal Classification (ASJC) codes

  • Immunology and Allergy
  • Immunology

Cite this

Ichikawa, A., Tachibana, H., Kawamoto, S., Kamei, M., Honjoh, T., Hashizume, S., & Shirahata, S. (1997). Cytokeratin 8 and 19 as antigens recognized by adenocarcinoma-reactive human monoclonal antibody AE6F4. Human Antibodies, 8(4), 195-202.

Cytokeratin 8 and 19 as antigens recognized by adenocarcinoma-reactive human monoclonal antibody AE6F4. / Ichikawa, Akira; Tachibana, Hirofumi; Kawamoto, Seiji; Kamei, Masanori; Honjoh, Tsutomu; Hashizume, Shuichi; Shirahata, Sanetaka.

In: Human Antibodies, Vol. 8, No. 4, 04.12.1997, p. 195-202.

Research output: Contribution to journalArticle

Ichikawa, A, Tachibana, H, Kawamoto, S, Kamei, M, Honjoh, T, Hashizume, S & Shirahata, S 1997, 'Cytokeratin 8 and 19 as antigens recognized by adenocarcinoma-reactive human monoclonal antibody AE6F4', Human Antibodies, vol. 8, no. 4, pp. 195-202.
Ichikawa A, Tachibana H, Kawamoto S, Kamei M, Honjoh T, Hashizume S et al. Cytokeratin 8 and 19 as antigens recognized by adenocarcinoma-reactive human monoclonal antibody AE6F4. Human Antibodies. 1997 Dec 4;8(4):195-202.
Ichikawa, Akira ; Tachibana, Hirofumi ; Kawamoto, Seiji ; Kamei, Masanori ; Honjoh, Tsutomu ; Hashizume, Shuichi ; Shirahata, Sanetaka. / Cytokeratin 8 and 19 as antigens recognized by adenocarcinoma-reactive human monoclonal antibody AE6F4. In: Human Antibodies. 1997 ; Vol. 8, No. 4. pp. 195-202.
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