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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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pI profiles across HC CDR3 regions of NZM2410-derived ANAs and non-ANAs. The mean pI value (isoelectric point) at each HC CDR3 position (from H95 to H100a) was calculated as detailed in Materials and Methods. As evident from Table III, few Abs had CDR3 residues beyond position H100a. The mean pI values observed among the NZM2410-derived ANAs (n = 49) and non-ANAs (n = 40) are plotted. Importantly, all clonal replicates have been removed from both databases. The pI values of the ANAs were systematically compared with the pI values of the non-ANAs at each indicated CDR position using the Student's t test (*, P < 0.05; **, P < 0.01).
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fig6: pI profiles across HC CDR3 regions of NZM2410-derived ANAs and non-ANAs. The mean pI value (isoelectric point) at each HC CDR3 position (from H95 to H100a) was calculated as detailed in Materials and Methods. As evident from Table III, few Abs had CDR3 residues beyond position H100a. The mean pI values observed among the NZM2410-derived ANAs (n = 49) and non-ANAs (n = 40) are plotted. Importantly, all clonal replicates have been removed from both databases. The pI values of the ANAs were systematically compared with the pI values of the non-ANAs at each indicated CDR position using the Student's t test (*, P < 0.05; **, P < 0.01).

Mentions: The CDR1, CDR2, and CDR3 sequences of NZM2410-derived ANA HC and LC are listed in Table III and Table IV, respectively. The HC CDR3 lengths were not significantly different between these two groups of Abs (9.5 residues among the ANAs vs. 9.9 residues among the non-ANAs; P > 0.05). Likewise, no differences were noted in LC CDR1 lengths (13.5 residues vs. 12.6; P > 0.05). Previous papers comparing ANAs and non-ANAs have observed the CDR3 regions of ANA HC to be significantly more cationic (for reviews see references 20–25). In resonance with those observations, the mean pI value (averaged across the first six residues) of HC CDR3 was significantly higher among the NZM2410-derived ANAs compared with the non-ANAs (6.18 vs. 5.76; P < 0.006; Fig. 6). Interestingly, the CDR3 regions of ANAs demonstrated alternating peaks of significant cationicity at positions H96 (P < 0.05), H98 (P < 0.01), and H100 (P < 0.05), compared with the non-ANAs. The intervening positions at H95, H97, and H99 appeared to be uniformly neutral (with mean pI values hovering just below 6). In contrast, the non-ANAs were relatively neutral at most of these CDR3 positions and significantly more anionic at H98 (Fig. 6).


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)

pI profiles across HC CDR3 regions of NZM2410-derived ANAs and non-ANAs. The mean pI value (isoelectric point) at each HC CDR3 position (from H95 to H100a) was calculated as detailed in Materials and Methods. As evident from Table III, few Abs had CDR3 residues beyond position H100a. The mean pI values observed among the NZM2410-derived ANAs (n = 49) and non-ANAs (n = 40) are plotted. Importantly, all clonal replicates have been removed from both databases. The pI values of the ANAs were systematically compared with the pI values of the non-ANAs at each indicated CDR position using the Student's t test (*, P < 0.05; **, P < 0.01).
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2211797&req=5

fig6: pI profiles across HC CDR3 regions of NZM2410-derived ANAs and non-ANAs. The mean pI value (isoelectric point) at each HC CDR3 position (from H95 to H100a) was calculated as detailed in Materials and Methods. As evident from Table III, few Abs had CDR3 residues beyond position H100a. The mean pI values observed among the NZM2410-derived ANAs (n = 49) and non-ANAs (n = 40) are plotted. Importantly, all clonal replicates have been removed from both databases. The pI values of the ANAs were systematically compared with the pI values of the non-ANAs at each indicated CDR position using the Student's t test (*, P < 0.05; **, P < 0.01).
Mentions: The CDR1, CDR2, and CDR3 sequences of NZM2410-derived ANA HC and LC are listed in Table III and Table IV, respectively. The HC CDR3 lengths were not significantly different between these two groups of Abs (9.5 residues among the ANAs vs. 9.9 residues among the non-ANAs; P > 0.05). Likewise, no differences were noted in LC CDR1 lengths (13.5 residues vs. 12.6; P > 0.05). Previous papers comparing ANAs and non-ANAs have observed the CDR3 regions of ANA HC to be significantly more cationic (for reviews see references 20–25). In resonance with those observations, the mean pI value (averaged across the first six residues) of HC CDR3 was significantly higher among the NZM2410-derived ANAs compared with the non-ANAs (6.18 vs. 5.76; P < 0.006; Fig. 6). Interestingly, the CDR3 regions of ANAs demonstrated alternating peaks of significant cationicity at positions H96 (P < 0.05), H98 (P < 0.01), and H100 (P < 0.05), compared with the non-ANAs. The intervening positions at H95, H97, and H99 appeared to be uniformly neutral (with mean pI values hovering just below 6). In contrast, the non-ANAs were relatively neutral at most of these CDR3 positions and significantly more anionic at H98 (Fig. 6).

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