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Lupus-Derived Human Monoclonal IgM anti-DNA Antibody Displays Monospecificity, High Affinity and Private Idiotype SpecificityImmunology Research Laboratory, St Luke's Hospital, 4400 Wornall Road, Kansas City, MO 64111, USA
Immunology Research Laboratory, St Luke's Hospital, 4400 Wornall Road, Kansas City, MO 64111, USA A human monoclonal IgM k anti-DNA antibody, designated 2F7, was prepared by somatic hybridization of peripheral blood lymphocytes from a lupus patient with a human-mouse heterohybridoma cell line, K6H6/B5. 2F7 was tested for its antigen binding and idiotypic specificity by direct binding and inhibition enzyme-linked immunosorbent assays. 2F7 had a high binding activity to single-stranded DNA (ssDNA) but not to double-stranded DNA. It cross-reacted with single-stranded homopolymers with pyrimidine bases and double-stranded polynucleotides containing those homopolymers, suggesting that 2F7 recognizes a conformational determinant made up of both deoxyribose-phosphate backbone and specific nucleotide base. 2F7 did not cross-react with eight structurally unrelated self-antigens. Dissociation constant (Kd) of 2F7 for sonicated ssDNA was approximately 4.5 x 10 -9 M, indicating its relatively high affinity. Idiotypic characterization with rabbit anti-idiotype raised against 2F7 suggested that 2F7 expressed an idiotype at or near its antigen-binding sites that was not detected in sera from 20 unrelated lupus patients, 10 lupus family members and 10 normal individuals. These results suggest that certain IgM class anti-DNA antibodies in human systemic lupus erythematosus may arise by antigen stimulation and not simply by polyclonal B-cell activation.
Key Words: Monoclonal • Anti-DNA • Monospecificity • High affinity • Private idiotype
Lupus, Vol. 1, No. 6,
363-368 (1992) |
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