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Scientific article
English

Diverse antigen specificity of erythrocyte-reactive monoclonal autoantibodies from NZB mice

Published inClinical and experimental immunology, vol. 105, no. 2, p. 313-320
Publication date1996
Abstract

The specificities of a panel of erythrocyte-reactive MoAbs derived from NZB mice with autoimmune haemolytic anaemia (AIHA) were determined by immunoprecipitation and immunoblotting. Of the eight antibodies, two (IgG1 MoAb 105-2H and IgG2a MoAb 34-3C) immunoprecipitated a 105-kD component identified as the erythrocyte anion channel band 3. A similar band was also immunoprecipitated by the IgG2b MoAb 34-2B when used at relatively high concentrations, but none of the remaining hybridoma antibodies precipitated any labelled erythrocyte components. In immunoblotting experiments only 34-2B reacted with band 3, indicating that the epitope recognized by this MoAb is robust and differs from the determinant(s) recognized by 105-2H and 34-3C. The remaining MoAbs to react by immunoblotting were the IgM antibodies IE10 and 4C8, both of which bound to a doublet corresponding to band 4.1 from the internal erythrocyte membrane skeleton. Of the three MoAbs which gave negative results in immunoprecipitation and immunoblotting, the IgM antibodies 103-7E and 106-10E reacted poorly with intact erythrocytes by flow cytometry, but the IgG1 antibody 31-9D bound well. ELISAs demonstrated that all four IgM MoAbs are polyreactive, since they bound to histones from a panel of nuclear antigens, and additionally 103-7E reacted with phosphatidyl choline. It is concluded that band 3 is an important autoantigen in NZB AIHA. However, since 3/5 haemolytic MoAbs failed to participate this antigen, either these antibodies represent minor components of the total autoantibody response, or responses to diverse possibly non-protein surface antigens also contribute to the pathogenesis of the disease.

Keywords
  • Anemia, Hemolytic, Autoimmune/ immunology
  • Animals
  • Anion Exchange Protein 1, Erythrocyte/analysis
  • Antibodies, Monoclonal/ immunology
  • Antibody Specificity
  • Autoantibodies/ immunology
  • Erythrocytes/ immunology
  • Flow Cytometry
  • Immunoblotting
  • Mice
  • Mice, Inbred NZB/ immunology
  • Phospholipids/immunology
  • Precipitin Tests
Affiliation Not a UNIGE publication
Citation (ISO format)
DE SA OLIVEIRA, G. G. et al. Diverse antigen specificity of erythrocyte-reactive monoclonal autoantibodies from NZB mice. In: Clinical and experimental immunology, 1996, vol. 105, n° 2, p. 313–320. doi: 10.1046/j.1365-2249.1996.d01-772.x
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ISSN of the journal0009-9104
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