Mouse monoclonal antibody against human soluble liver antigen and the detection of human antibodies possessing the same idiotype in patients with chronic liver diseases.
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概要
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A gene encoding the variable regions of the heavy and light chains of a mouse monoclonal antibody H2, which specifically reacts with human liver cells, was cloned in a phagemid vector. Anti-idiotypic antibody was produced by immunizing the original mouse antibody with various absorptions. The cloned phage specifically reacted with this anti-idiotypic antibody, and the reaction between the cloned phage and anti-idiotypic antibody was specifically blocked by the original antibody. The soluble protein, expressed as a fusion protein, was detected as a 30 kDa single band on sodium dodecyl sulfate polyacrylamide gel electrophoresis. This protein was demonstrated to bind specifically to an anti H2 idiotypic antibody by Western blot analysis.<BR>The sera of patients with various diseases were assayed using this anti-idiotypic antibody and sandwich enzyme-linked immunosorbent assay. Only the sera of patients with chronic liver diseases highly reacted and this binding was specifically attenuated by the cloned soluble protein. This liverspecific idiotype-bearing antibody detected in the serum of a patient with chronic hepatitis C could be purified and this antibody mediated antibody-dependent cytotoxicity against a hepatocyte derived cell line Chang <I>in vitro</I>.<BR>These findings suggest that a human antibody that reacts with human liver cells, can be detected in patients with chronic liver diseases and can contribute to the liver cell damage.
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