Measurement of antibodies to double-stranded and single-stranded DNA using bacterio phage Ď DNA as the antigen \ Avidity of antibodies to DNA in pati
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1 Measurement of antibodies to double-stranded and single-stranded DNA using bacterio phage Ď DNA as the antigen \ Avidity of antibodies to DNA in patients with lupus nephritis \ M0T0AKI SAIT0, YASUYUKI UECHI, KIMIMASA NAKABAYASHI and TOSHIHIK0 NAGASAWA The 1st Department of Internal Medicine, Kyorin University School of Medicine, Tokyo, Japan Key words : bacteriophage Ď DNA, IgG anti-dsdna antibodies, IgG anti-ssdna antibodies, high avidity, lupus nephritis IgG-class antibodies to double-stranded (ds) and single-stranded (ss) DNA in patients with collagen disease were measured by enzyme-linked immunosorbent assay (ELISA) using bacteriophage Ď DNA as the antigen. Moreover, avidity of IgG-class antibodies to both dsdna and ssdna in patients with systemic lupus erythematosus (SLE) was evaluated by ELISA as salt-dependent binding activities. The results are as follows:1) Elevated IgG-class antibodies to dsdna were largely restricted to patients with active SLE. Furthermore, the levels of IgG-class antibodies to dsdna correlated with the disease activity of SLE. 2) Lupus nephritis (LN) patients with nephrotic syndrome (NS) or diffuse proliferative LN as deter mined by renal biopsy had high levels of IgG-class antibodies to both dsdna and ssdna. 3) LN patients with NS showed the highest means of avidity index of IgG-class antibodies to both dsdna and ssdna. These findings suggest that the present method of measuring IgG-class antibodies to dsdna is useful for the diagnosis and management of patients with active SLE and that IgG-class antibodies to both dsdna and ssdna with high avidity may be an important factor in the pathogenesis of LN with NS.
2 Table 1. Patients. ( 30
3 Fig. 1. Titers of IgG anti-dsdna antibody in patients with collagen diseases. Fig. 2. Titers of IgG anti-ssdna antibody in patients with collagen diseases. ( 31
4 Table 2. Correlation between titiers of IgG anti-ds/ssdna antibody and anti-dna antibody measured by various methods in patients with SLE. Fig. 3. Disease activity of SLE and change of titers of IgG anti-ds/ssdna antibody. ( 32
5 Fig. 4. Renal disorders and titiers of IgG anti-ds / ssdna antibody in patients with SLE. Fig. 5. Renal histological findings and titiers of IgG anti-ds / ssdna antibody inpatients with LN. ( 33
6 BacteriophageλDNAを 1108 は,208.9±119.4IU/ml,IgG抗ssDNA抗 に お け 価 お,IgG抗ssDNA抗 るIgG抗 体 価 は,70.7±111.8IU/ml,IgG抗ssDNA NaCI濃 抗 体 価 は,288.2±257.3AU/mlで,IgG抗dsDNA抗 よ MLN群 びIgG抗ssDNA抗 ー プ ス 腎 炎 患 に,DPLN群 体 を 測 定 dsdna抗 者 に お け る 高 塩 濃 度下 応 系 の塩濃 度 を上 示 indexは,反 が175mM.225mM,275mMの Avidity analysis concentrations of in 応 系 のNaCI濃 の 時,NS腎 症 群 が 非NS腎 体 のavidity 度 が225mMお よ び275mM 症 群 お よ び非 腎 症 群 に比 べ した 考 察 症 群 で は 各 々, 今 回 我 々 は,新 DNAを 時,NS腎 anti-dsdna patients 行 な い,本 反 応 系 し く 開 発 さ れ たbacteriophageλ 抗 原 と し て 固 相 化 し たELISAキ I gg抗dsdna抗 indexは IgG indexは,反 度 腎症 よ び275mMの あ り,IgG抗ssDNA抗 有意 (p<0.05)に高値を示 症 群 で 体 のavidity 度 が225mMお NaCl 症 症 群 で は 各 々,79.8±16.1,67.2±21.8,56.7±24.3%,非腎症群では各々,77.1±19.8, ±26.5%で 群 で は 各 々,79.3±15.3,63.6±18.4,46.6±19.5%で 6. 応 系 の 時,NS腎 NS腎 し た IgG抗 は 各 々,92.7±5.7,84.0±.15.0,71.6±24.5%,非 Fig. 度 が175mM 225mM,275mMの が 昇 さ せ て 抗 93.6±8.8,83.7±16.6,77.0±24.1%,非NS腎 のNaGl濃 indexは,反 群 で は 各 々,91.4±8.6,87.2±14.1,83,0±17.3%,非 で の 抗 応 系 のNaCl濃 時,NS腎 あ り,IgG抗dsDNA抗 体 のavidity し た 一方 高 値 を 示 し た し た 成 績 をFig.6,7に 体 のavidity 高 値 を 示 体 体 本 キ ッ ト に お い て,反 DNA抗 は 共 に 比 べ 有 意(p<0.05)に (4)ル DNA抗 体 価 体 の測 定 が 非 腎 症 群 に 比 べ 有 意(p<0.05)に 体 佃1は, 513.2±247.7AU/mlで,MLN群 dsdna抗 抗 原 と し た 抗DNA抗 体 とIgG抗ssDNA抗 症 群 Fig. by 7. Avidity NaCl with SLE. SLE. ( 34 体 の同時測定を キ ッ トの 有 用 性 と,SLE特 者に お け るIgG抗dsDNA抗 ッ トを 用 い に ル ー プ ス 腎 炎,患 体,IgG抗ssDNA抗 analysis concentrations of IgG in antiss-dna patients 体の by with
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9 immune cmplexes in serum and glomeruli of pa tients with systemic lupus erythematosus Am J Pathol74: , ) Izui S, Lambert PH, Fournie GJ, Tuner H, Miesher 1) Schur PH, Sandson J: Immunologic factors and clinical activity in systemic lupus erythematosus N Engl J Med 278: , ) Koffler D, Carr RI, Agnello V, Thoburn B, Kunkel HG: Antibodies to polynucleotides in human sera: antigenic specificty and relation to disease J Exp Med 134: , ) Lange A: Evaluation of the simultaneous estimation of anti-dsdna and anti-ssdna antibodies clinical purpose Clin Exp Immunol 31: , 1978 for 5) Sasaki T, Kadono T, Endo F, Seliguchi Y, Saito T, Arakawa M, Furuyama T, Yoshinaga K: Relation ship between complement-fixing (hemolytic) anti bodies to single-stranded and double-stranded DNA and the prognosis in systemic lupus erythem atosus Tohoku J Exp Med 138: , ) Koffler D, Agnello V, Kunkel HG: Polynucleotide PA: Features of systemic lupus erythematosus in mice injected bacterial lipopolysaccharides. Identi fication of circulation DNA and renal localization of DNA-anti-DNAcomplexes J Rxp Med 145: , 9) Winfield JB, Faifeman I, Koffler: Avidity of anti- DNA antibodies in serum and IgG glomerular elu ates from a patient with systemic lupus erythemat osus J Clin Invest 59: 90-96, ) Leon SA, Green A, Ehrlich GE, Poland M, Shapipo B: Avidity of antibodies in SLE, Relation to sever ity of renal involvement Arthritis Rheum 20: 23-29, ) Mcgrath Jr H, Biundo Jr JJ: A longitudinal study of high and low avidity antibodies to double stranded DNA in systemic lupus erythematosus Arthritis Rheum 28: , ) Yoshida M, Yoshida H, Muso E, Kawai C, Hamas hima Y: Avidity of anti-dna antibodies and its clinical significance in patients with SLE Jim Arch Int Med 34: 89-96, ) Smeenk RJT, Rooijen AV, Swaak TJG: Dissocia tion studies of DNA/anti-DNA complexes inrela tion to anti-dna avidity J Immunol Methods 109: 27-35, ) Urowitz MB, Gladma DG, Tozman ECS, Gold smith CH: The lupus activity criteria count ( 37
10 (LACC) The Journal of Rheumatology 11: , ) Rothfield MF, Stollar BD: The relation of im munorelobulin class, pattern of antinuclear anti body, and complement-fixing antibodies to DNA in sera from patients with systemic lupus erythemat osus J Clin Invest 46: , ) Tojo T, Friou GJ: Lupus nephritis: varying com plement-fixing properties of immunoglobulin G antibodies to antigens of cell nuclei Science 161: , ) Muryoi T, Sasaki T, Hatakeyama A, Shibata S, Suzuki M, Seino J, Yoshinaga K: Clonotypes of anti-dna antibodies expressing specific idiotypes in immune complexes of patients with active lupus nephritis J Immunol 144: , 1990
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