Antigen-binding diversity of human hybridoma autoantibodies derived from splenocytes of patients with SLE.
The antigen-binding specificity of human hybridoma-derived monoclonal autoantibodies (mAb) was analysed with mAbs derived from the spleens of two patients with active systemic lupus erythematosus (SLE). From one patient 72 mAbs (RSP clones) and from the other 173 mAbs (RT clones) were obtained. The binding specificity of these mAbs was analysed by solid- and fluid-phase ELISA against the autoantigens ssDNA, dsDNA, cardiolipin, SmRNP, histones, Sm-D and SS-B (La) synthetic peptides, and foreign antigens including bacterial polysaccharides. In addition, antinuclear antibody activity and anti-dsDNA binding were confirmed by fluorescence staining methods. Reflecting the patient's serological profile, none of the antibodies from the RSP clones reacted with ssDNA or dsDNA but 12 reacted with cardiolipin. In addition, three mAbs reacted with H4, five with U1 RNP, two with Sm-D peptides and 12 with SS-B peptides. In contrast, from the RT fusion, nine mAbs reacted with ssDNA, HI and SS-B peptides, seven with cardiolipin, four with dsDNA, two with Sm-D peptides and one each with H2A, H3 and H4. In many cases one mAb showed reactivity with more than one antigen: for example, mAb RT 72 binds to ssDNA, dsDNA, cardiolipin, H1, H4 and an Sm-D peptide; RT 6 binds to H1, SmRNP and ubiquitinated histone H2A. However, none of the antibodies showed 'across the board' polyreactivity; indeed, the selectivity of the reactions was notable and marked variation in antibody affinity was recorded. Eight of the mAbs bound to Salmonella typhimurium and two to the Klebsiella polysaccharide K-30.(ABSTRACT TRUNCATED AT 250 WORDS)
Ravirajan
C T
CT
Department of Rheumatology Research, University College and Middlesex School of Medicine, London, UK.
Kalsi
J
J
Wiloch
H W
HW
Barakat
S
S
Tuaillon
N
N
Irvine
W
W
Cockayne
A
A
Harris
A
A
Williams
D G
DG
Williams
W
W
D000293:Adolescent; D000328:Adult; D017153:Antibodies, Anticardiolipin / Q000378:metabolism; D000974:Antibodies, Antinuclear / Q000378:metabolism; D000911:Antibodies, Monoclonal; D000915:Antibody Affinity; D000916:Antibody Diversity; D000918:Antibody Specificity; D001323:Autoantibodies / Q000378:metabolism*; D001324:Autoantigens; D005260:Female; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D007074:Immunoglobulin G / Q000378:metabolism; D007075:Immunoglobulin M / Q000378:metabolism; D008180:Lupus Erythematosus, Systemic / Q000276:immunology*; D013154:Spleen / Q000276:immunology
D017153:Antibodies, Anticardiolipin; D000974:Antibodies, Antinuclear; D000911:Antibodies, Monoclonal; D001323:Autoantibodies; D001324:Autoantigens; D007074:Immunoglobulin G; D007075:Immunoglobulin M