A single-stranded DNA-cross-reactive immunogenic epitope of human homocysteine-inducible endoplasmic reticulum protein

Scand J Immunol. 2011 Sep;74(3):296-303. doi: 10.1111/j.1365-3083.2011.02572.x.

Abstract

The mechanism involved in generating anti-DNA antibodies (Abs) remains unclear, as DNA is poorly immunogenic. Molecular mimicry between DNA and non-DNA substances has been implicated as a possible mechanism. We previously reported that homocysteine-inducible endoplasmic reticulum protein (Herp), which is induced by endoplasmic reticulum stress, is recognized by anti-double-stranded DNA (dsDNA) IgG from patients with systemic lupus erythematosus and that immunization with Herp elicits anti-dsDNA Abs in BALB/c mice. In this study, we observed that anti-single-stranded DNA (ssDNA) Abs were also generated in Herp-immunized BALB/c mice and established an anti-Herp monoclonal antibody (mAb), HT4, which specifically cross-reacted with ssDNA. The epitope of the HT4 mAb on Herp, 'EPAGSNR', was identified by screening a synthetic peptide library. The binding of the HT4 mAb to the peptide was competitively inhibited by ssDNA. Immunization of the epitope peptide elicited anti-ssDNA Abs in BALB/c mice. These results indicate that the epitope exists in a human self-protein, mimics ssDNA and shows antigenicity for anti-ssDNA Abs in normal mice. Anti-ssDNA Abs are often found in patients with drug-induced lupus erythematosus. Treatment with representative drugs that cause drug-induced lupus (chlorpromazine, procainamide and hydralazine) induced Herp expression and apoptosis in HeLa cells. These findings suggest that molecular mimicry between Herp and ssDNA is involved in anti-ssDNA Ab production in drug-induced lupus.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Antinuclear / biosynthesis
  • Antibodies, Antinuclear / immunology*
  • Antibodies, Monoclonal / immunology
  • Apoptosis
  • Autoantibodies / immunology*
  • Cell Line, Tumor
  • Cross Reactions
  • DNA / immunology*
  • DNA, Single-Stranded / immunology*
  • Enzyme-Linked Immunosorbent Assay
  • Epitope Mapping
  • Epitopes / immunology
  • HeLa Cells
  • Homocysteine / immunology
  • Humans
  • Immunoglobulin G / immunology
  • Lupus Erythematosus, Systemic / chemically induced*
  • Lupus Erythematosus, Systemic / immunology
  • Membrane Proteins / biosynthesis
  • Membrane Proteins / genetics
  • Membrane Proteins / immunology*
  • Mice
  • Mice, Inbred BALB C
  • Microscopy, Fluorescence

Substances

  • Antibodies, Antinuclear
  • Antibodies, Monoclonal
  • Autoantibodies
  • DNA, Single-Stranded
  • Epitopes
  • HERPUD1 protein, human
  • Immunoglobulin G
  • Membrane Proteins
  • Homocysteine
  • DNA