Repertoire cloning of lupus anti-DNA autoantibodies.
To investigate the autoantibody repertoire associated with SLE, we have created phage display IgG Fab libraries from two clinically active SLE patients and from the healthy identical twin of one of these patients. The libraries from the lupus discordant twins were found to both include unusually large representations of the V(H)5 gene family. By panning with DNA, the SLE libraries each yielded IgG anti-double-stranded (ds) DNA autoantibodies, which are characteristic of lupus disease. These included a V(H)5 autoantibody from the affected twin, that has a targeted cluster of mutations that potentially improves binding affinity. The recovered IgG anti-dsDNA autoantibodies expressed the same idiotypes associated with the in vivo IgG anti-dsDNA response of the respective SLE donor. Heavy-light chain shuffling experiments demonstrated a case in which the in vitro creation of anti-dsDNA binding activity required restrictive pairing of a heavy chain with Vlambda light chains similar to those in circulating anti-dsDNA autoantibodies. By contrast, IgG anti-ds autoantibodies could not be recovered from the library from the healthy twin, or from shuffled libraries with heavy chains from the healthy twin. These repertoire analyses illustrate how inheritance and somatic processes interplay to produce lupus-associated IgG autoantibodies.
Roben
P
P
The Sam and Rose Stein Institute for Research on Aging and the Theodore Gildred Cancer Center, Department of Medicine, University of California, San Diego, La Jolla 92093, USA.
Barbas
S M
SM
Sandoval
L
L
Lecerf
J M
JM
Stollar
B D
BD
Solomon
A
A
Silverman
G J
GJ
D000595:Amino Acid Sequence; D000974:Antibodies, Antinuclear / Q000276:immunology*; D001323:Autoantibodies / Q000276:immunology*; D015551:Autoimmunity / Q000276:immunology*; D001402:B-Lymphocytes; D003001:Cloning, Molecular; D004247:DNA / Q000276:immunology / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000276:immunology / Q000378:metabolism; D005260:Female; D005796:Genes; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000276:immunology; D007130:Immunoglobulin Idiotypes / Q000235:genetics / Q000276:immunology; D007135:Immunoglobulin Variable Region / Q000276:immunology; D008180:Lupus Erythematosus, Systemic / Q000276:immunology*; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D012699:Serology; D014430:Twins, Monozygotic
D000974:Antibodies, Antinuclear; D001323:Autoantibodies; D004268:DNA-Binding Proteins; D007140:Immunoglobulin Fab Fragments; D007130:Immunoglobulin Idiotypes; D007135:Immunoglobulin Variable Region; D011994:Recombinant Proteins; D004247:DNA