Anti-DNA antibody mediated catalysis is isotype dependent

Yumin Xia, Ertan Eryilmaz, Qiuting Zhang, David Cowburn, Chaim Putterman

Research output: Contribution to journalArticlepeer-review

12 Scopus citations

Abstract

Anti-DNA antibodies are the serological hallmark of systemic lupus erythematosus, and participate in the pathogenesis of lupus nephritis by cross-reacting with multiple renal antigens. Previously, using a panel of murine anti-DNA IgGs that share identical variable regions but that differ in the constant regions, we demonstrated that the cross-reaction and renal pathogenicity of anti-DNA antibodies are isotype dependent. In this study, we investigated the catalytic potential of this anti-DNA antibody panel, and determined its isotype dependency. The three isotype switch variants (IgG1, IgG2a, IgG2b) and the parent IgG3 PL9-11 anti-DNA antibodies were compared in their catalysis of 500 base pair linear double stranded DNA and a 12-mer peptide (ALWPPNLHAWVP), by gel analysis, MALDI-TOF mass spectrometry, and nuclear magnetic resonance spectroscopy. The binding affinity of anti-DNA antibodies to double stranded DNA and peptide antigens were assessed by ELISA and surface plasmon resonance. We found that the PL9-11 antibody isotypes vary significantly in their potential to catalyze the cleavage of both linear and double stranded DNA and the proteolysis of peptides. The degree of the cleavage and proteolysis increases with the incubation temperature and time. While different PL9-11 isotypes have the same initial attack sites within the ALWPPNLHAWVP peptide, there was no correlation between binding affinity to the peptide and proteolysis rates. In conclusion, the catalytic properties of anti-DNA antibodies are isotype dependent. This finding provides further evidence that antibodies that share the same variable region, but which have different constant regions, are functionally distinct. The catalytic effects modulated by antibody constant regions need to be considered in the design of therapeutic antibodies (abzymes) and peptides designed to block pathogenic autoantibodies.

Original languageEnglish
Pages (from-to)33-43
Number of pages11
JournalMolecular Immunology
Volume69
DOIs
StatePublished - 1 Jan 2016
Externally publishedYes

Bibliographical note

Publisher Copyright:
© 2015 Elsevier Ltd.

Funding

This work was supported by grants from the NIH (AR048692 and DK090319 to C. Putterman and OD016305 to D. Cowburn). We thank Dr. Huiyong Cheng (Dept. of Biochemistry, Albert Einstein College of Medicine) for help with the Biacore analysis, and Dr. Jennifer Aguilan (Dept. of Biochemistry, Albert Einstein College of Medicine) for help with MALDI-TOF mass spectrometry.

FundersFunder number
National Institutes of HealthS10OD016305, DK090319
National Institute of Arthritis and Musculoskeletal and Skin DiseasesR56AR048692

    Keywords

    • Anti-DNA antibody
    • Catalysis
    • Systemic lupus erythematosus

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