Influence of variable domain glycosylation on anti-neutrophil cytoplasmic autoantibodies and anti-glomerular basement membrane autoantibodies

BMC Immunology

Ming-Hui Zhao¹  Min Chen¹  Xiao-Yu Jia¹  Zhao Cui¹  Xiao-Wei Yang¹  Shen-Ju Gou¹  Peng-Cheng Xu² 

[1] Renal Division, Department of Medicine, Peking University First Hospital, Institute of Nephrology, Peking University, Key Laboratory of Renal Disease, Ministry of Health of China, Beijing 100034, China;Department of Nephrology, General Hospital of Tianjin Medical University, Tianjin 300052, China

关键词: Anti-GBM;    ANCA;    Variable region;    Glycosylation;   

Others  :  1077938
DOI  :  10.1186/1471-2172-13-10

 received in 2011-09-28, accepted in 2012-03-09,  发布年份 2012

【 摘 要 】

Background

The pathophysiological significance of variable region glycosylation of autoantibodies is still unclear. In the current study, the influence of the variable region N-linked oligosaccharides on the reactivity of three autoantibody specificities was investigated with Sambucus nigra agglutinin (SNA), which mainly binds to oligosaccharides with terminal α2, 6-linked sialic acid on the variable region of IgG.

Methods

Twenty-seven patients with serum positive anti-neutrophil cytoplasmic autoantibodies (ANCA) against myeploperoxidase (MPO) or proteinase 3 (PR3), or autoantibodies against glomerular basement membrane (GBM) were included. Total IgG was isolated and separated into non-SNA-binding and SNA-binding fractions with SNA affinity chromatography. Antigen-specific IgG was purified by immunoaffinity chromatography.

Results

At the same concentration of IgG, the antigen binding level of non-SNA-binding IgG was significantly lower than that of SNA-binding IgG for MPO-ANCA (absorbance value at 405 nm, 0.572 ± 0.590 vs. 0.962 ± 0.670, P < 0.001) and for PR3-ANCA (0.362 ± 0.530 vs. 0.560 ± 0.531, P = 0.003). The antigen binding level of non-SNA-binding IgG was significantly higher than that of SNA-binding IgG for anti-GBM antibodies (1.301 ± 0.594 vs. 1.172 ± 0.583, P = 0.044). The level of variable region glycosylation of total IgG was significantly lower than that of affinity-purified MPO-ANCA (1.021 ± 0.201 vs. 1.434 ± 0.134, P = 0.004). The level of variable region glycosylation of total IgG was significantly higher than that of affinity-purified anti-GBM antibodies (1.034 ± 0.340 vs. 0.734 ± 0.333, P = 0.007). The SNA-binding fraction of MPO-ANCA-containing IgG and PR3-ANCA-containing IgG induced higher levels of neutrophil oxygen radical production than the corresponding non-SNA-binding fractions (P < 0.001 and P = 0.043, respectively). The level of variable region glycosylation of affinity-purified MPO-ANCA was higher in active AAV than the same patients in remission (P = 0.001).

Conclusion

Characteristics of variable region glycosylation of ANCA and anti-GBM antibodies were different from that of total IgG, which might influence the antigen-binding ability of these antibodies. Variable region glycosylation of ANCA might influence the effect of ANCA-induced neutrophils respiratory burst.

【 授权许可】

   
2012 Xu et al; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

Figure 5.

Figure 6.

【 参考文献 】

• [1]Jennette JC, Falk RJ: Small-vessel vasculitis. N Engl J Med 1997, 337:1512-1523.
• [2]Short AK, Esnault VL, Lockwood CM: Anti-neutrophil cytoplasm antibodies and anti-glomerular basement membrane antibodies: two coexisting distinct autoreactivities detectable in patients with rapidly progressive glomerulonephritis. Am J Kidney Dis 1995, 26:439-445.
• [3]Hellmark T, Niles JL, Collins AB, McCluskey RT, Brunmark C: Comparison of anti-GBM antibodies in sera with or without ANCA. J Am Soc Nephrol 1997, 8:376-385.
• [4]Parekh RB, Dwek RA, Sutton BJ, et al.: Association of rheumatoid arthritis and primary osteoarthritis with changes in the glycosylation pattern of total serum IgG. Nature 1985, 316:452-457.
• [5]Takahashi N, Ishii I, Ishihara H, et al.: Comparative structural study of the N-linked oligosaccharides of human normal and pathological immunoglobulin G. Biochemistry 1987, 26:1137-1144.
• [6]Holland M, Takada K, Okumoto T, et al.: Hypogalactosylation of serum IgG in patients with ANCA-associated systemic vasculitis. Clin Exp Immunol 2002, 129:183-190.
• [7]Espy C, Morelle W, Kavian N, et al.: Sialylation levels of anti-proteinase 3 antibodies are associated with the activity of granulomatosis with polyangiitis (Wegener's). Arthritis Rheum 2011, 63:2105-2115.
• [8]Donadel G, Calabro A, Sigounas G, et al.: Human polyreactive and monoreactive antibodies: effect of glycosylation on antigen binding. Glycobiology 1994, 4:491-496.
• [9]Rudd PM, Leatherbarrow RJ, Rademacher TW, Dwek RA: Diversification of the IgG molecule by oligosaccharides. Mol Immunol 1991, 28:1369-1378.
• [10]Huang L, Biolsi S, Bales KR, Kuchibhotla U: Impact of variable domain glycosylation on antibody clearance: an LC/MS characterization. Anal Biochem 2006, 349:197-207.
• [11]Malhotra R, Wormald MR, Rudd PM, et al.: Glycosylation changes of IgG associated with rheumatoid arthritis can activate complement via the mannose-binding protein. Nat Med 1995, 1:237-243.
• [12]van Timmeren MM, van der Veen BS, Stegeman CA, et al.: IgG glycan hydrolysis attenuates ANCA-mediated glomerulonephritis. J Am Soc Nephrol 2010, 21:1103-1114.
• [13]Kusakabe K, Kanaya K, Sato S, Kato M, Mochizuki K, et al.: Accumulation enhancement of human monoclonal antibody HB4C5 to lung tumor xenografts by N-deglycosylation. J Nucl Med 1994, 35:289-295.
• [14]Co MS, Scheinberg DA, Avdalovic NM, et al.: Genetically engineered deglycosylation of the variable domain increases the affinity of an anti-CD33 monoclonal antibody. Mol Immunol 1993, 30:1361-1367.
• [15]Kato M, Mochizuki K, Hashizume S, et al.: Activity enhancement of a lung cancer-associated human monoclonal antibody HB4C5 by N-deglycosylation. Hum Antibodies Hybridomas 1993, 4:9-14.
• [16]Matsuuchi L, Sharon J, Morrison SL: An analysis of heavy chain glycopeptides of hybridoma antibodies: correlation between antibody specificity and sialic acid content. J Immunol 1981, 127:2188-2190.
• [17]Tachibana H, Shirahata S, Murakami H: Generation of specificity-variant antibodies by alteration of carbohydrate in light chain of human monoclonal antibodies. Biochem Biophys Res Commun 1992, 189:625-632.
• [18]Coloma MJ, Trinh RK, Martinez AR, Morrison SL: Position effects of variable region carbohydrate on the affinity and in vivo behavior of an anti-(1- > 6) dextran antibody. J Immunol 1999, 162:2162-2170.
• [19]Shibuya N, Goldstein IJ, Broekaert WF, et al.: The elderberry (Sambucus nigra L.) bark lectin recognizes the Neu5Ac(alpha 2-6)Gal/GalNAc sequence. J Biol Chem 1987, 262:1596-1601.
• [20]Stadlmann J, Weber A, Pabst M, et al.: A close look at human IgG sialylation and subclass distribution after lectin fractionation. Proteomics 2009, 9:4143-4153.
• [21]Dalziel M, McFarlane I, Axford JS: Lectin analysis of human immunoglobulin G N-glycan sialylation. Glycoconj J 1999, 16:801-807.
• [22]Rademacher TW: Network theory of glycosylation-etiologic and pathogenic implications of changes in IgG glycoform levels in autoimmunity. Semin Cell Biol 1991, 2:327-337.
• [23]Holland M, Yagi H, Takahashi N, et al.: Differential glycosylation of polyclonal IgG, IgG-Fc and IgG-Fab isolated from the sera of patients with ANCA-associated systemic vasculitis. Biochim Biophys Acta 2006, 1760:669-677.
• [24]Guhr T, Bloem J, Derksen NI: Enrichment of sialylated IgG by lectin fractionation does not enhance the efficacy of immunoglobulin G in a murine model of immune thrombocytopenia. PLoS One 2011, 6:e21246.
• [25]Endo T, Wright A, Morrison SL, Kobata A: Glycosylation of the variable region of immunoglobulin G--site specific maturation of the sugar chains. Mol Immunol 1995, 32:931-940.
• [26]Kinoshita N, Ohno M, Nishiura T, et al.: Glycosylation at the Fab portion of myeloma immunoglobulin G and increased fucosylated biantennary sugar chains: structural analysis by high-performance liquid chromatography and antibody-lectin enzyme immunoassay using Lens culinaris agglutinin. Cancer Res 1991, 51:5888-5892.
• [27]Jennette JC, Falk RJ, Andrassy K, et al.: Nomenclature of systemic vasculitides. Proposal of an international consensus conference. Arthritis Rheum 1994, 37:187-192.
• [28]Zhao MH, Lockwood CM: A comprehensive method to purify three major ANCA antigens: proteinase 3, myeloperoxidase and bactericidal/permeability-increasing protein from human neutrophil granule acid extract. J Immunol Methods 1996, 197:121-130.
• [29]Cui Z, Zhao MH, Xin G, Wang HY: Characteristics and Prognosis of Chinese Patients with Anti-Glomerular Basement Membrane Disease. Nephron Clin Pract 2005, 99:c49-c55.
• [30]Franco Fraguas L, Carlsson J, Lönnberg M: Lectin affinity chromatography as a tool to differentiate endogenous and recombinant erythropoietins. J Chromatogr A 2008, 1212:82-88.
• [31]Mimura Y, Ashton PR, Takahashi N, Harvey DJ, Jefferis R: Contrasting glycosylation profiles between Fab and Fc of a human IgG protein studied by electrospray ionization mass spectrometry. J Immunol Methods 2007, 326:116-126.
• [32]Xu PC, Chen M, Cui Z, Zhao MH: Influence of myeloperoxidase by anti-myeloperoxidase antibodies and its association with the disease activity in microscopic polyangiitis. Rheumatology (Oxford) 2010, 49:2068-2075.
• [33]Kettritz R, Schreiber A, Luft FC, Haller H: Role of mitogen-activated protein kinases in activation of human neutrophils by antineutrophil cytoplasmic antibodies. J Am Soc Nephrol 2001, 12:37-46.
• [34]Borza DB, Netzer KO, Leinonen A, et al.: The goodpasture autoantigen. Identification of multiple cryptic epitopes on the NC1 domain of the alpha3(IV) collagen chain. J Biol Chem 2000, 275:6030-6037.
• [35]Yang R, Hellmark T, Zhao J, et al.: Antigen and epitope specificity of anti-glomerular basement membrane antibodies in patients with goodpasture disease with or without anti-neutrophil cytoplasmic antibodies. J Am Soc Nephrol 2007, 18:1338-1343.
• [36]Luqmani RA, Bacon PA, Moots RJ, et al.: Birmingham Vasculitis Activity Score (BVAS) in systemic necrotizing vasculitis. QJM 1994, 87:671-678.
• [37]Zhang Z, Pan H, Chen X: Mass spectrometry for structural characterization of therapeutic antibodies. Mass Spectrom Rev 2009, 28:147-176.
• [38]Zhu D, McCarthy H, Ottensmeier CH, et al.: Acquisition of potential N-glycosylation sites in the immunoglobulin variable region by somatic mutation is a distinctive feature of follicular lymphoma. Blood 2002, 99:2562-2568.
• [39]Derry CJ, Dunn MJ, Rees AJ, Pusey CD: Restricted specificity of the autoantibody response in Goodpasture's syndrome demonstrated by two-dimensional western blotting. Clin Exp Immunol 1991, 86:457-463.
• [40]Malle E, Buch T, Grone HJ: Myeloperoxidase in kidney disease. Kidney Int 2003, 64:1956-1967.
• [41]Franssen CF, Stegeman CA, Kallenberg CG, et al.: Antiproteinase 3-and antimyeloperoxidase-associated vasculitis. Kidney Int 2000, 57:2195-2206.
• [42]Gala FA, Morrison SL: V region carbohydrate and antibody expression. J Immunol 2004, 172:5489-5494.
• [43]Berek C: The development of B cells and the B-cell repertoire in the microenvironment of the germinal center. Immunol Rev 1992, 126:5-19.
• [44]Tomlinson IM, Walter G, Jones PT, et al.: The imprint of somatic hypermutation on the repertoire of human germline V genes. J Mol Biol 1996, 256:813-817.
• [45]Cox JP, Tomlinson IM, Winter G: A directory of human germ-line V kappa segments reveals a strong bias in their usage. Eur J Immunol 1994, 24:827-836.
• [46]Walter G, Tomlinson IM, Dear PH, et al.: Comparison of the human germline and rearranged VH repertoire reveals complementarity between germline variability and somatic mutation. Ann N Y Acad Sci 1995, 764:180-182.
• [47]Dunn-Walters D, Boursier L, Spencer J: Effect of somatic hypermutation on potential N-glycosylation sites in human immunoglobulin heavy chain variable regions. Mol Immunol 2000, 37:107-113.