Limits...
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.

Show MeSH

Related in: MedlinePlus

Jk usage frequencies among ANAs and non-ANAs. Indicated are the LC Jk usage frequencies of NZM2410-derived ANAs (n = 46), NZM2410-derived non-ANAs (n = 40), previously documented ANAs (from NCBI/GenBank/EMBL/DDBJ; n = 264), and non-ANAs (from NCBI/GenBank/EMBL/DDBJ; n = 145). The NCBI collection of ANAs and non-ANAs represents two new databases of LC sequences recently constructed and analyzed. The control ANAs represent 264 previously documented ANA LC sequences, drawn from 35 primary works, from which clonal replicates have been removed; they consisted of 139 anti-ssDNA, 103 anti-dsDNA, and 22 antinucleosome Abs (26). The NCBI/GenBank/EMBL/DDBJ “non-ANAs” represent the LC sequences of 145 non-ANAs (with known antigen specificities) drawn from the NCBI/GenBank/EMBL/DDBJ database, with no overlapping target antigen specificities. Importantly, all clonal replicates have been removed from all four of the databases studied, so as to minimize any potential bias due to multi-member clones. The frequencies of Jk5 among the NZM2410-derived ANAs and non-ANAs were significantly higher (P < 0.013 and P < 0.009, respectively) when compared with the corresponding frequencies observed among the NCBI-derived ANAs and non-ANAs.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC2211797&req=5

fig5: Jk usage frequencies among ANAs and non-ANAs. Indicated are the LC Jk usage frequencies of NZM2410-derived ANAs (n = 46), NZM2410-derived non-ANAs (n = 40), previously documented ANAs (from NCBI/GenBank/EMBL/DDBJ; n = 264), and non-ANAs (from NCBI/GenBank/EMBL/DDBJ; n = 145). The NCBI collection of ANAs and non-ANAs represents two new databases of LC sequences recently constructed and analyzed. The control ANAs represent 264 previously documented ANA LC sequences, drawn from 35 primary works, from which clonal replicates have been removed; they consisted of 139 anti-ssDNA, 103 anti-dsDNA, and 22 antinucleosome Abs (26). The NCBI/GenBank/EMBL/DDBJ “non-ANAs” represent the LC sequences of 145 non-ANAs (with known antigen specificities) drawn from the NCBI/GenBank/EMBL/DDBJ database, with no overlapping target antigen specificities. Importantly, all clonal replicates have been removed from all four of the databases studied, so as to minimize any potential bias due to multi-member clones. The frequencies of Jk5 among the NZM2410-derived ANAs and non-ANAs were significantly higher (P < 0.013 and P < 0.009, respectively) when compared with the corresponding frequencies observed among the NCBI-derived ANAs and non-ANAs.

Mentions: Fig. 5 depicts the Jk utilization profiles of the NZM2410-derived ANAs and non-ANAs, as well as those of ANAs and non-ANAs described previously (26). Although the ANAs and non-ANAs derived from the NZM2410 mice did not differ significantly in terms of their Jk usage, both these groups of Abs had significantly higher Jk5 utilization frequencies. Therefore, the Jk5:Jk1 ratios in the NZM2410-derived ANAs (ratio = 1.2), and non-ANAs (ratio = 1.1), were significantly higher (P < 0.013 and P < 0.006, respectively) compared with the ANAs (ratio = 0.6) and non-ANAs (ratio = 0.4) drawn from the National Center for Biotechnology Information/GenBank/EMBL/DDBJ databases. Factoring in the nuclear antigen fine specificities of the ANAs did not reveal any further differences. Thus, it is tempting to posit that these NZM2410-derived Abs (i.e., both the ANAs as well as the non-ANAs) may largely be the end product of a vigorous receptor-editing program at the LC locus. Nevertheless, one cannot exclude the possibility that other, yet to be defined, immunogenetic factors may be influencing Jk recombination in the NZM2410 genetic background.


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)

Jk usage frequencies among ANAs and non-ANAs. Indicated are the LC Jk usage frequencies of NZM2410-derived ANAs (n = 46), NZM2410-derived non-ANAs (n = 40), previously documented ANAs (from NCBI/GenBank/EMBL/DDBJ; n = 264), and non-ANAs (from NCBI/GenBank/EMBL/DDBJ; n = 145). The NCBI collection of ANAs and non-ANAs represents two new databases of LC sequences recently constructed and analyzed. The control ANAs represent 264 previously documented ANA LC sequences, drawn from 35 primary works, from which clonal replicates have been removed; they consisted of 139 anti-ssDNA, 103 anti-dsDNA, and 22 antinucleosome Abs (26). The NCBI/GenBank/EMBL/DDBJ “non-ANAs” represent the LC sequences of 145 non-ANAs (with known antigen specificities) drawn from the NCBI/GenBank/EMBL/DDBJ database, with no overlapping target antigen specificities. Importantly, all clonal replicates have been removed from all four of the databases studied, so as to minimize any potential bias due to multi-member clones. The frequencies of Jk5 among the NZM2410-derived ANAs and non-ANAs were significantly higher (P < 0.013 and P < 0.009, respectively) when compared with the corresponding frequencies observed among the NCBI-derived ANAs and non-ANAs.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2211797&req=5

fig5: Jk usage frequencies among ANAs and non-ANAs. Indicated are the LC Jk usage frequencies of NZM2410-derived ANAs (n = 46), NZM2410-derived non-ANAs (n = 40), previously documented ANAs (from NCBI/GenBank/EMBL/DDBJ; n = 264), and non-ANAs (from NCBI/GenBank/EMBL/DDBJ; n = 145). The NCBI collection of ANAs and non-ANAs represents two new databases of LC sequences recently constructed and analyzed. The control ANAs represent 264 previously documented ANA LC sequences, drawn from 35 primary works, from which clonal replicates have been removed; they consisted of 139 anti-ssDNA, 103 anti-dsDNA, and 22 antinucleosome Abs (26). The NCBI/GenBank/EMBL/DDBJ “non-ANAs” represent the LC sequences of 145 non-ANAs (with known antigen specificities) drawn from the NCBI/GenBank/EMBL/DDBJ database, with no overlapping target antigen specificities. Importantly, all clonal replicates have been removed from all four of the databases studied, so as to minimize any potential bias due to multi-member clones. The frequencies of Jk5 among the NZM2410-derived ANAs and non-ANAs were significantly higher (P < 0.013 and P < 0.009, respectively) when compared with the corresponding frequencies observed among the NCBI-derived ANAs and non-ANAs.
Mentions: Fig. 5 depicts the Jk utilization profiles of the NZM2410-derived ANAs and non-ANAs, as well as those of ANAs and non-ANAs described previously (26). Although the ANAs and non-ANAs derived from the NZM2410 mice did not differ significantly in terms of their Jk usage, both these groups of Abs had significantly higher Jk5 utilization frequencies. Therefore, the Jk5:Jk1 ratios in the NZM2410-derived ANAs (ratio = 1.2), and non-ANAs (ratio = 1.1), were significantly higher (P < 0.013 and P < 0.006, respectively) compared with the ANAs (ratio = 0.6) and non-ANAs (ratio = 0.4) drawn from the National Center for Biotechnology Information/GenBank/EMBL/DDBJ databases. Factoring in the nuclear antigen fine specificities of the ANAs did not reveal any further differences. Thus, it is tempting to posit that these NZM2410-derived Abs (i.e., both the ANAs as well as the non-ANAs) may largely be the end product of a vigorous receptor-editing program at the LC locus. Nevertheless, one cannot exclude the possibility that other, yet to be defined, immunogenetic factors may be influencing Jk recombination in the NZM2410 genetic background.

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