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Pathogenic profiles and molecular signatures of antinuclear autoantibodies rescued from NZM2410 lupus mice.

Liang Z, Xie C, Chen C, Kreska D, Hsu K, Li L, Zhou XJ, Mohan C - J. Exp. Med. (2004)

Bottom Line: Most intriguingly, the CDR3 regions of the ANAs exhibited alternating arginine/lysine peaks at H96, H98, and H100, with neutral troughs at H95, H97, and H99.To summarize, glomerular-binding anti-dsDNA antibodies appear to be the most pathogenic variety of lupus autoantibodies.The presence of an alternating charge pattern in their HC CDR3 regions appears to be a prominent hallmark of ANAs.

View Article: PubMed Central - PubMed

Affiliation: Simmons Arthritis Research Center, University of Texas Southwestern Medical School, Dallas 75390, USA.

ABSTRACT
Two outstanding questions concerning antinuclear antibodies (ANAs) in lupus involve their pathogenic potential and their molecular signatures. To address these questions, a panel of 56 antinuclear and 47 nonnuclear binding monoclonal antibodies was rescued from four seropositive NZM2410 lupus mice. The monoclonals varied in their reactivity to nucleosomes, ssDNA, dsDNA, and glomerular substrate. A large fraction of the antibodies demonstrated apparent polyreactivity (to DNA, histones, and glomerular antigens) due to bound, DNase-1 sensitive nuclear antigenic bridges. Although nephrophilic immunoglobulin (Ig) M and IgG antibodies were the most pathogenic, the dsDNA-binding antibodies were modestly so; in contrast, antinucleosome antibodies were clearly not pathogenic. Compared with the nonnuclear antigen-binding monoclonal antibodies rescued from the same mice, ANAs exhibited increased utilization of VH5/7183 genes and highly cationic heavy chain (HC) CDR3 regions. Most intriguingly, the CDR3 regions of the ANAs exhibited alternating arginine/lysine peaks at H96, H98, and H100, with neutral troughs at H95, H97, and H99. To summarize, glomerular-binding anti-dsDNA antibodies appear to be the most pathogenic variety of lupus autoantibodies. The presence of an alternating charge pattern in their HC CDR3 regions appears to be a prominent hallmark of ANAs.

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Somatic variations between members of antibody clones. 11 NZM2410-derived monoclonal ANAs are depicted that belonged to five independent clones, A–E, as indicated in Tables I, III, and IV. These clones have been labeled the same way as in Table I, except that the mouse origin identifiers (e.g., “ZA”, “ZB,” etc.) have been omitted. DsDNA-reactive clones are shaded in gray, whereas exclusive nucleosome binders are left unshaded. (top) The HC somatic mutations relative to the closest HC germline gene (indicated in oval labels with dotted borders) are depicted. (bottom) The LC somatic mutations relative to the closest LC germline gene (indicated in oval-shaped labels with dotted borders) are depicted. Thus, in clone “B,” mAbs 7H10 and 3F7 vary from the J558 germline gene, V23, by 6 or 10 residues, respectively, whereas they both bear the same unmutated LC Vk1 germline gene, bb1. Interestingly, whereas the first-listed two members of Clone D, 7F11 and 2D7, possessed a mutated RF Vk germline gene, the third member possessed an entirely different LC gene that differed from the Vk9 germline gene, ba9, by nine somatic mutations (not depicted).
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fig8: Somatic variations between members of antibody clones. 11 NZM2410-derived monoclonal ANAs are depicted that belonged to five independent clones, A–E, as indicated in Tables I, III, and IV. These clones have been labeled the same way as in Table I, except that the mouse origin identifiers (e.g., “ZA”, “ZB,” etc.) have been omitted. DsDNA-reactive clones are shaded in gray, whereas exclusive nucleosome binders are left unshaded. (top) The HC somatic mutations relative to the closest HC germline gene (indicated in oval labels with dotted borders) are depicted. (bottom) The LC somatic mutations relative to the closest LC germline gene (indicated in oval-shaped labels with dotted borders) are depicted. Thus, in clone “B,” mAbs 7H10 and 3F7 vary from the J558 germline gene, V23, by 6 or 10 residues, respectively, whereas they both bear the same unmutated LC Vk1 germline gene, bb1. Interestingly, whereas the first-listed two members of Clone D, 7F11 and 2D7, possessed a mutated RF Vk germline gene, the third member possessed an entirely different LC gene that differed from the Vk9 germline gene, ba9, by nine somatic mutations (not depicted).

Mentions: Finally, we examined the multimember clonal families among the rescued ANAs. Of the six clonal families observed, five consisted of two to three nonidentical members each (Fig. 8), whereas the remaining clone (Table I, clone “f”) consisted of two identical isolates. This was consistent with the mutation frequencies listed in Table VI, whereas the HC of these clones displayed high mutation frequencies, particularly in the CDR regions, the LC varied little from the germline sequences, for the most part. In contrast to clones “A,” “B,” and “E,” clones “C” and “D” exhibited intraclonal differences in ANA fine specificity. Because both members of clone C possessed the same germline-encoded VH1/J558 10B10S germline gene, the few LC mutations exhibited by clone 4E2 must have been responsible for the dsDNA reactivity profile of this clone. In this context, the replacement mutation at L27e, which results in an R residue, is particularly attractive. This is perhaps the first example of an instance where somatic mutation of the LC partner may have been responsible for generating a dsDNA binder, beginning with a nucleosome-restricted precursor.


Pathogenic profiles and molecular signatures of antinuclear autoantibodies rescued from NZM2410 lupus mice.

Liang Z, Xie C, Chen C, Kreska D, Hsu K, Li L, Zhou XJ, Mohan C - J. Exp. Med. (2004)

Somatic variations between members of antibody clones. 11 NZM2410-derived monoclonal ANAs are depicted that belonged to five independent clones, A–E, as indicated in Tables I, III, and IV. These clones have been labeled the same way as in Table I, except that the mouse origin identifiers (e.g., “ZA”, “ZB,” etc.) have been omitted. DsDNA-reactive clones are shaded in gray, whereas exclusive nucleosome binders are left unshaded. (top) The HC somatic mutations relative to the closest HC germline gene (indicated in oval labels with dotted borders) are depicted. (bottom) The LC somatic mutations relative to the closest LC germline gene (indicated in oval-shaped labels with dotted borders) are depicted. Thus, in clone “B,” mAbs 7H10 and 3F7 vary from the J558 germline gene, V23, by 6 or 10 residues, respectively, whereas they both bear the same unmutated LC Vk1 germline gene, bb1. Interestingly, whereas the first-listed two members of Clone D, 7F11 and 2D7, possessed a mutated RF Vk germline gene, the third member possessed an entirely different LC gene that differed from the Vk9 germline gene, ba9, by nine somatic mutations (not depicted).
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Related In: Results  -  Collection

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fig8: Somatic variations between members of antibody clones. 11 NZM2410-derived monoclonal ANAs are depicted that belonged to five independent clones, A–E, as indicated in Tables I, III, and IV. These clones have been labeled the same way as in Table I, except that the mouse origin identifiers (e.g., “ZA”, “ZB,” etc.) have been omitted. DsDNA-reactive clones are shaded in gray, whereas exclusive nucleosome binders are left unshaded. (top) The HC somatic mutations relative to the closest HC germline gene (indicated in oval labels with dotted borders) are depicted. (bottom) The LC somatic mutations relative to the closest LC germline gene (indicated in oval-shaped labels with dotted borders) are depicted. Thus, in clone “B,” mAbs 7H10 and 3F7 vary from the J558 germline gene, V23, by 6 or 10 residues, respectively, whereas they both bear the same unmutated LC Vk1 germline gene, bb1. Interestingly, whereas the first-listed two members of Clone D, 7F11 and 2D7, possessed a mutated RF Vk germline gene, the third member possessed an entirely different LC gene that differed from the Vk9 germline gene, ba9, by nine somatic mutations (not depicted).
Mentions: Finally, we examined the multimember clonal families among the rescued ANAs. Of the six clonal families observed, five consisted of two to three nonidentical members each (Fig. 8), whereas the remaining clone (Table I, clone “f”) consisted of two identical isolates. This was consistent with the mutation frequencies listed in Table VI, whereas the HC of these clones displayed high mutation frequencies, particularly in the CDR regions, the LC varied little from the germline sequences, for the most part. In contrast to clones “A,” “B,” and “E,” clones “C” and “D” exhibited intraclonal differences in ANA fine specificity. Because both members of clone C possessed the same germline-encoded VH1/J558 10B10S germline gene, the few LC mutations exhibited by clone 4E2 must have been responsible for the dsDNA reactivity profile of this clone. In this context, the replacement mutation at L27e, which results in an R residue, is particularly attractive. This is perhaps the first example of an instance where somatic mutation of the LC partner may have been responsible for generating a dsDNA binder, beginning with a nucleosome-restricted precursor.

Bottom Line: Most intriguingly, the CDR3 regions of the ANAs exhibited alternating arginine/lysine peaks at H96, H98, and H100, with neutral troughs at H95, H97, and H99.To summarize, glomerular-binding anti-dsDNA antibodies appear to be the most pathogenic variety of lupus autoantibodies.The presence of an alternating charge pattern in their HC CDR3 regions appears to be a prominent hallmark of ANAs.

View Article: PubMed Central - PubMed

Affiliation: Simmons Arthritis Research Center, University of Texas Southwestern Medical School, Dallas 75390, USA.

ABSTRACT
Two outstanding questions concerning antinuclear antibodies (ANAs) in lupus involve their pathogenic potential and their molecular signatures. To address these questions, a panel of 56 antinuclear and 47 nonnuclear binding monoclonal antibodies was rescued from four seropositive NZM2410 lupus mice. The monoclonals varied in their reactivity to nucleosomes, ssDNA, dsDNA, and glomerular substrate. A large fraction of the antibodies demonstrated apparent polyreactivity (to DNA, histones, and glomerular antigens) due to bound, DNase-1 sensitive nuclear antigenic bridges. Although nephrophilic immunoglobulin (Ig) M and IgG antibodies were the most pathogenic, the dsDNA-binding antibodies were modestly so; in contrast, antinucleosome antibodies were clearly not pathogenic. Compared with the nonnuclear antigen-binding monoclonal antibodies rescued from the same mice, ANAs exhibited increased utilization of VH5/7183 genes and highly cationic heavy chain (HC) CDR3 regions. Most intriguingly, the CDR3 regions of the ANAs exhibited alternating arginine/lysine peaks at H96, H98, and H100, with neutral troughs at H95, H97, and H99. To summarize, glomerular-binding anti-dsDNA antibodies appear to be the most pathogenic variety of lupus autoantibodies. The presence of an alternating charge pattern in their HC CDR3 regions appears to be a prominent hallmark of ANAs.

Show MeSH
Related in: MedlinePlus