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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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The distribution of “R/K” and “D/E” residues in the HC CDR3 regions of ANAs and non-ANAs. In A, the respective frequencies of R/K and D/E residues at each of the HC CDR3 positions, H95– H100a, among the NZM2410-derived ANAs (n = 49) and non-ANAs (n = 40) are depicted. Differences (between ANAs and non-ANAs) that attained statistical significance are denoted (*, P < 0.05; **, P < 0.01). The solid line arrowhead indicates the alternating frequency peaks of cationicity between H96 and H98 of ANAs; the dotted line arrowhead indicates the alternating peaks of anionicity between H96 and H98 among the non-ANAs. There were too few sequences with CDR3 positions extending beyond H100a (Table III). In B, a similar analysis was performed using previously reported ANAs (n = 269) and non-ANAs (n = 3,600) (for review see reference 25). These control ANAs represent 269 previously documented ANAs drawn from 35 primary works, from which clonal replicates have been removed; they consisted of 143 anti-ssDNA, 103 anti-dsDNA, and 23 antinucleosome Abs (25). The control “non-ANAs” represent the HCs of all Abs deposited in the Kabat database. Differences that attained statistical significance are denoted (*, P < 0.05; **, P < 0.01; ***, P < 0.001).
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fig7: The distribution of “R/K” and “D/E” residues in the HC CDR3 regions of ANAs and non-ANAs. In A, the respective frequencies of R/K and D/E residues at each of the HC CDR3 positions, H95– H100a, among the NZM2410-derived ANAs (n = 49) and non-ANAs (n = 40) are depicted. Differences (between ANAs and non-ANAs) that attained statistical significance are denoted (*, P < 0.05; **, P < 0.01). The solid line arrowhead indicates the alternating frequency peaks of cationicity between H96 and H98 of ANAs; the dotted line arrowhead indicates the alternating peaks of anionicity between H96 and H98 among the non-ANAs. There were too few sequences with CDR3 positions extending beyond H100a (Table III). In B, a similar analysis was performed using previously reported ANAs (n = 269) and non-ANAs (n = 3,600) (for review see reference 25). These control ANAs represent 269 previously documented ANAs drawn from 35 primary works, from which clonal replicates have been removed; they consisted of 143 anti-ssDNA, 103 anti-dsDNA, and 23 antinucleosome Abs (25). The control “non-ANAs” represent the HCs of all Abs deposited in the Kabat database. Differences that attained statistical significance are denoted (*, P < 0.05; **, P < 0.01; ***, P < 0.001).

Mentions: Consistent with the pI profile differences portrayed in Fig. 6, NZM2410-derived ANAs exhibited significantly more “R” residues (9.4 vs. 3.8%, representing the mean percentages averaged across H95-H100a; P < 0.02) and significantly less “D” residues (5.8 vs. 10.5%; P < 0.05) in their HC CDR3 regions, compared with the non-ANAs. As is clear from Fig. 7 A, the NZM2410-derived ANAs exhibited peak usage of “R/K” residues (especially “R”) at the alternating positions, H96, H98, and H100 (as well as H100a). In the intervening positions, H95, H97, and H99, the frequencies of R/K residues were outweighed by the corresponding frequencies of “D/E” residues. In contrast with the ANAs, the non-ANAs exhibited peak usage of anionic D/E residues (especially “D”) at the same alternating positions, H96, H98, and H100. These reciprocal charge patterns at H96–H98 are highlighted using arrowheads in Fig. 7 A; these findings resonate well with the pI profile differences shown in 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)

The distribution of “R/K” and “D/E” residues in the HC CDR3 regions of ANAs and non-ANAs. In A, the respective frequencies of R/K and D/E residues at each of the HC CDR3 positions, H95– H100a, among the NZM2410-derived ANAs (n = 49) and non-ANAs (n = 40) are depicted. Differences (between ANAs and non-ANAs) that attained statistical significance are denoted (*, P < 0.05; **, P < 0.01). The solid line arrowhead indicates the alternating frequency peaks of cationicity between H96 and H98 of ANAs; the dotted line arrowhead indicates the alternating peaks of anionicity between H96 and H98 among the non-ANAs. There were too few sequences with CDR3 positions extending beyond H100a (Table III). In B, a similar analysis was performed using previously reported ANAs (n = 269) and non-ANAs (n = 3,600) (for review see reference 25). These control ANAs represent 269 previously documented ANAs drawn from 35 primary works, from which clonal replicates have been removed; they consisted of 143 anti-ssDNA, 103 anti-dsDNA, and 23 antinucleosome Abs (25). The control “non-ANAs” represent the HCs of all Abs deposited in the Kabat database. Differences that attained statistical significance are denoted (*, P < 0.05; **, P < 0.01; ***, P < 0.001).
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fig7: The distribution of “R/K” and “D/E” residues in the HC CDR3 regions of ANAs and non-ANAs. In A, the respective frequencies of R/K and D/E residues at each of the HC CDR3 positions, H95– H100a, among the NZM2410-derived ANAs (n = 49) and non-ANAs (n = 40) are depicted. Differences (between ANAs and non-ANAs) that attained statistical significance are denoted (*, P < 0.05; **, P < 0.01). The solid line arrowhead indicates the alternating frequency peaks of cationicity between H96 and H98 of ANAs; the dotted line arrowhead indicates the alternating peaks of anionicity between H96 and H98 among the non-ANAs. There were too few sequences with CDR3 positions extending beyond H100a (Table III). In B, a similar analysis was performed using previously reported ANAs (n = 269) and non-ANAs (n = 3,600) (for review see reference 25). These control ANAs represent 269 previously documented ANAs drawn from 35 primary works, from which clonal replicates have been removed; they consisted of 143 anti-ssDNA, 103 anti-dsDNA, and 23 antinucleosome Abs (25). The control “non-ANAs” represent the HCs of all Abs deposited in the Kabat database. Differences that attained statistical significance are denoted (*, P < 0.05; **, P < 0.01; ***, P < 0.001).
Mentions: Consistent with the pI profile differences portrayed in Fig. 6, NZM2410-derived ANAs exhibited significantly more “R” residues (9.4 vs. 3.8%, representing the mean percentages averaged across H95-H100a; P < 0.02) and significantly less “D” residues (5.8 vs. 10.5%; P < 0.05) in their HC CDR3 regions, compared with the non-ANAs. As is clear from Fig. 7 A, the NZM2410-derived ANAs exhibited peak usage of “R/K” residues (especially “R”) at the alternating positions, H96, H98, and H100 (as well as H100a). In the intervening positions, H95, H97, and H99, the frequencies of R/K residues were outweighed by the corresponding frequencies of “D/E” residues. In contrast with the ANAs, the non-ANAs exhibited peak usage of anionic D/E residues (especially “D”) at the same alternating positions, H96, H98, and H100. These reciprocal charge patterns at H96–H98 are highlighted using arrowheads in Fig. 7 A; these findings resonate well with the pI profile differences shown in 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