Cloning of variable regions of an antibody that reacts with the soluble fraction of human liver cells and its possible value in chronic liver disease

Hidetsugu Saito, Shinichiro Tada, Hirotoshi Ebinuma, Kazuhiro Atsukawa, Tetsuya Masuda, Yasutaka Inagaki, Kanji Tsuchimoto, Toshio Morizane, Hiromasa Ishii

Research output: Contribution to journalArticle

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Abstract

A gene encoding the variable regions of the heavy and light chains of a mouse monoclonal antibody designated H2, which specifically reacts with human liver cells, was cloned into a phagemid vector. The clone of the variable region was designed to be expressed as a separate protein, the structure of which is the same as that of the mouse antibody. The cloned phage protein specifically reacted with anti-idiotypic antibodies produced in rabbit against the original mouse antibody, and this reaction was specifically blocked by the original antibody. The soluble protein, expressed as a fusion protein, was detected as a single 30-kd band on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and specifically bound to an anti-H2 idiotypic antibody as determined by Western blot analysis. Sera of patients with various disease were assayed for antibodies to anti-H2 by sandwich enzyme-linked immunosorbent assay (ELISA). Only sera from patients with chronic liver disease reacted strongly. This binding was specifically blocked by the cloned soluble protein. The nucleotide sequences of the variable regions were determined by the dideoxy chain-termination method, and the sequences were approximately 95% identical to those of other mouse immunoglobulins. This findings suggest that a human antibody with the same idiotype as a mouse monoclonal antibody that reacts with human liver cells, can be detected in patients with chronic liver disese, suggeting that autoimmunity may be partly reponsoble for these disease.

Original languageEnglish
Pages (from-to)1498-1506
Number of pages9
JournalHepatology
Volume23
Issue number6
DOIs
Publication statusPublished - 1996

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Liver Diseases
Organism Cloning
Chronic Disease
Antibodies
Liver
Anti-Idiotypic Antibodies
Proteins
Monoclonal Antibodies
Serum
Autoimmunity
Sodium Dodecyl Sulfate
Bacteriophages
Immunoglobulins
Polyacrylamide Gel Electrophoresis
Clone Cells
Western Blotting
Enzyme-Linked Immunosorbent Assay
Rabbits
Light
Genes

ASJC Scopus subject areas

  • Hepatology

Cite this

Cloning of variable regions of an antibody that reacts with the soluble fraction of human liver cells and its possible value in chronic liver disease. / Saito, Hidetsugu; Tada, Shinichiro; Ebinuma, Hirotoshi; Atsukawa, Kazuhiro; Masuda, Tetsuya; Inagaki, Yasutaka; Tsuchimoto, Kanji; Morizane, Toshio; Ishii, Hiromasa.

In: Hepatology, Vol. 23, No. 6, 1996, p. 1498-1506.

Research output: Contribution to journalArticle

Saito, Hidetsugu ; Tada, Shinichiro ; Ebinuma, Hirotoshi ; Atsukawa, Kazuhiro ; Masuda, Tetsuya ; Inagaki, Yasutaka ; Tsuchimoto, Kanji ; Morizane, Toshio ; Ishii, Hiromasa. / Cloning of variable regions of an antibody that reacts with the soluble fraction of human liver cells and its possible value in chronic liver disease. In: Hepatology. 1996 ; Vol. 23, No. 6. pp. 1498-1506.
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