The constant region contributes to the antigenic specificity and renal pathogenicity of murine anti-DNA antibodies

Yumin Xia, Rahul D. Pawar, Antonio S. Nakouzi, Leal Herlitz, Anna R. Broder, Kui Liu, Beatrice Goilav, Manxia Fan, Ling Wang, Quan Zhen Li, Arturo Casadevall, Chaim Putterman

Research output: Contribution to journalArticle

33 Citations (Scopus)

Abstract

Affinity for DNA and cross-reactivity with renal antigens are associated with enhanced renal pathogenicity of lupus autoantibodies. In addition, certain IgG subclasses are enriched in nephritic kidneys, suggesting that isotype may determine the outcome of antibody binding to renal antigens. To investigate if the isotype of DNA antibodies affects renal pathogenicity by influencing antigen binding, we derived IgM, IgG1, IgG2b and IgG2a forms of the PL9-11 antibody (IgG3 anti-DNA) by in vitro class switching or PCR cloning. The affinity and specificity of PL9-11 antibodies for nuclear and renal antigens were analyzed using ELISA, Western blotting, surface plasmon resonance (SPR), binding to mesangial cells, and glomerular proteome arrays. Renal deposition and pathogenicity were assayed in mice injected with PL9-11 hybridomas. We found that PL9-11 and its isotype-switched variants had differential binding to DNA and chromatin (IgG3 > IgG2a > IgG1 > IgG2b > IgM) by direct and competition ELISA, and SPR. In contrast, in binding to laminin and collagen IV the IgG2a isotype actually had the highest affinity. Differences in affinity of PL9-11 antibodies for renal antigens were mirrored in analysis of specificity for glomeruli, and were associated with significant differences in renal pathogenicity in vivo and survival. Our novel findings indicate that the constant region plays an important role in the nephritogenicity of antibodies to DNA by affecting immunoglobulin affinity and specificity. Increased binding to multiple glomerular and/or nuclear antigens may contribute to the renal pathogenicity of anti-DNA antibodies of the IgG2a and IgG3 isotype. Finally, class switch recombination may be another mechanism by which B cell autoreactivity is generated.

Original languageEnglish (US)
Pages (from-to)398-411
Number of pages14
JournalJournal of Autoimmunity
Volume39
Issue number4
DOIs
StatePublished - Dec 2012

Fingerprint

Antinuclear Antibodies
Virulence
Epitopes
Kidney
Immunoglobulin G
Antibodies
Antigens
Nuclear Antigens
Surface Plasmon Resonance
DNA
Immunoglobulin M
Enzyme-Linked Immunosorbent Assay
Immunoglobulin Class Switching
Mesangial Cells
Hybridomas
Laminin
Proteome
Autoantibodies
Genetic Recombination
Chromatin

Keywords

  • Anti-DNA antibodies
  • Isotype switching
  • Lupus nephritis
  • Systemic lupus erythematosus (SLE)

ASJC Scopus subject areas

  • Immunology
  • Immunology and Allergy

Cite this

The constant region contributes to the antigenic specificity and renal pathogenicity of murine anti-DNA antibodies. / Xia, Yumin; Pawar, Rahul D.; Nakouzi, Antonio S.; Herlitz, Leal; Broder, Anna R.; Liu, Kui; Goilav, Beatrice; Fan, Manxia; Wang, Ling; Li, Quan Zhen; Casadevall, Arturo; Putterman, Chaim.

In: Journal of Autoimmunity, Vol. 39, No. 4, 12.2012, p. 398-411.

Research output: Contribution to journalArticle

Xia, Yumin ; Pawar, Rahul D. ; Nakouzi, Antonio S. ; Herlitz, Leal ; Broder, Anna R. ; Liu, Kui ; Goilav, Beatrice ; Fan, Manxia ; Wang, Ling ; Li, Quan Zhen ; Casadevall, Arturo ; Putterman, Chaim. / The constant region contributes to the antigenic specificity and renal pathogenicity of murine anti-DNA antibodies. In: Journal of Autoimmunity. 2012 ; Vol. 39, No. 4. pp. 398-411.
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