Dual reactivity of several monoclonal anti-nucleosome autoantibodies for double-stranded DNA and a short segment of histone H3.
We have shown previously that four IgG monoclonal autoantibodies (mAbs) reacted in ELISA with both double-stranded (ds) DNA and peptide 83-100 of histone H3. The peptide 83-100 contains a cysteine residue at position 96 and readily dimerizes at pH 7-8. We describe here that only the 83-100 dimers, and not the 83-100 monomers, are recognized by the four antibodies and inhibit in ELISA the binding of mAbs to dsDNA. The equilibrium affinity constants (Ka) and kinetic rate constants of two of these mAbs were measured in a biosensor system. Ka values were significantly higher when these mAbs were tested with dsDNA as compared with the 83-100 dimer. Further higher Ka values were measured with mononucleosomes containing DNA and histones. It is proposed that these four mAbs are directed against a topographic determinant formed by DNA and the region 83-100 of H3 present as a dimer at the surface of nucleosome, and that they react, although significantly less well, with DNA and peptide dimer tested separately. This study provides a quantitative and kinetic basis to interaction between several antibodies and distinct antigenic structures and allows us to better understand the structural basis of apparent autoantibody cross-reactivity.
Stemmer
C
C
Institut de Biologie Moléculaire et Cellulaire, UPR 9021 CNRS, 15 rue Descartes, 67000 Strasbourg, France.
Richalet-Sécordel
P
P
van Bruggen
M
M
Kramers
K
K
Berden
J
J
Muller
S
S
D000595:Amino Acid Sequence; D000818:Animals; D000974:Antibodies, Antinuclear / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D003429:Cross Reactions; D004247:DNA / Q000276:immunology*; D006657:Histones / Q000737:chemistry / Q000276:immunology*; D007700:Kinetics; D051379:Mice; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry
D000974:Antibodies, Antinuclear; D000911:Antibodies, Monoclonal; D006657:Histones; D010446:Peptide Fragments; D004247:DNA