Mimotopes of polyreactive anti-DNA antibodies identified using phage-display peptide libraries.
Three monoclonal IgG2a anti-DNA polyreactive autoantibodies, derived from lupus-prone mice (NZB x NZW)F1, were studied by surface plasmon resonance (BIAcore) analysis using three different synthetic double-stranded (ds) oligonucleotides of 25, 30, and 25 base pairs (bp). These monoclonal antibodies (mAb) exhibited dissociation rate constants (k(off)), ranging from 0.0001 (mAb F14.6 and F4.1) to 0.01/s (mAb J20.8) and k(on) ranging from 2 x 10(5) to 2 x 10(6) /M/s. The screening of a constrained random peptide library displayed on M13 bacteriophages on these mAb allowed the determination of the specific consensus motifs (mimotopes) for mAb F14.6 and J20.8, but not for mAb F4.1. No cross-reaction was observed between F14.6- and J20.8-specific peptides (and vice versa). Binding of all phages selected on F14.6 was inhibited with 700 ng/ml soluble DNA. The binding of one group of peptides selected on J20.8 was inhibited by 400 ng/ml soluble DNA, of a second group by 2500 ng/ml, while binding of a third group could not be inhibited. The determined consensus sequences do not match with known sequences. Peptides specific for F14.6 share negative charges and aromatic rings that may mimic a DNA backbone, while peptides selected with J20.8 do not bear any negative charge, implying a different kind of molecular recognition, for example hydrogen or salt bonds. The peptides selected on J20.8 also bind serum antibodies from human patients with systemic lupus erythematosus. In addition, BALB/c mice immunized with some of the selected phages exhibit high serum titers of IgG3 anti-dsDNA antibodies, further supporting the hypothesis that peptide epitopes may mimic an oligonucleotide structure.
Sibille
P
P
Immunopharmacologie Moléculaire, Institut Cochin de Génétique Moléculaire, Paris, France.
Ternynck
T
T
Nato
F
F
Buttin
G
G
Strosberg
D
D
Avrameas
A
A
D000595:Amino Acid Sequence; D000818:Animals; D000974:Antibodies, Antinuclear / Q000096:biosynthesis / Q000737:chemistry* / Q000276:immunology; D000937:Antigen-Antibody Reactions / Q000187:drug effects; D017104:Bacteriophage M13 / Q000737:chemistry* / Q000378:metabolism; D001666:Binding Sites, Antibody / Q000187:drug effects; D001667:Binding, Competitive / Q000276:immunology; D004247:DNA / Q000276:immunology*; D000939:Epitopes / Q000276:immunology*; D015723:Gene Library; D006801:Humans; D007700:Kinetics; D008180:Lupus Erythematosus, Systemic / Q000097:blood / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D008814:Mice, Inbred NZB; D008969:Molecular Sequence Data; D009841:Oligonucleotides / Q000276:immunology; D010452:Peptide Biosynthesis; D010455:Peptides / Q000276:immunology* / Q000494:pharmacology
D000974:Antibodies, Antinuclear; D000939:Epitopes; D009841:Oligonucleotides; D010455:Peptides; D004247:DNA