Evidence is lacking for direct pathogenicity of human anti-proteinase-3 (PR3) antibodies in development of systemic vasculitis and granulomatosis with polyangiitis (GPA, Wegener's granulomatosis). Progress in study of these antibodies in rodents has been hampered by lack of PR3 expression on murine neutrophils, and by different Fc-receptor affinities for IgG across species. Therefore, we tested whether human anti-PR3 antibodies can induce acute vasculitis in mice with a human immune system. Chimeric mice were generated by injecting human haematopoietic stem cells into irradiated NOD-scid-IL2Rγ？/？ mice. Matched chimera mice were treated with human IgG from patients with: anti-PR3 positive renal and lung vasculitis; patients with non-vasculitic renal disease; or healthy controls. Six-days later, 39% of anti-PR3 treated mice had haematuria, compared with none of controls. There was punctate bleeding on the surface of lungs of anti-PR3 treated animals, with histological evidence of vasculitis and haemorrhage. Anti-PR3 treated mice had mild pauci-immune proliferative glomerulonephritis, with infiltration of human and mouse leukocytes. In 3 mice (17%) more severe glomerular injury was present. There were no glomerular changes in controls. Human IgG from patients with anti-PR3 autoantibodies is therefore pathogenic. This model of anti-PR3 antibody-mediated vasculitis may be useful in dissecting mechanisms of microvascular injury.
References
[1]
van der Woude FJ, Rasmussen N, Lobatto S, Wiik A, Permin H, et al. (1985) Autoantibodies against neutrophils and monocytes: tool for diagnosis and marker of disease activity in Wegener's granulomatosis. Lancet 1(8426): 425–9.
[2]
Falk RJ, Jennette JC (1988) Anti-neutrophil cytoplasmic autoantibodies with specificity for myeloperoxidase in patients with systemic vasculitis and idiopathic necrotizing and crescentic glomerulonephritis. N Engl J Med 318: 1651–7.
[3]
Niles JL, McCluskey RT, Ahmad MF, Arnaout MA (1989) Wegener's granulomatosis autoantigen is a novel neutrophil serine proteinase. Blood 74: 1888–93.
[4]
Jennette JC, Hoidal JR, Falk RJ (1990) Specificity of anti-neutrophil cytoplasmic autoantibodies for proteinase 3. Blood 75: 2263–4.
[5]
Goldschmeding R, van der Schoot CE, ten Bokkel Huinink D, Hack CE, van den Ende ME, et al. (1989) Wegener's granulomatosis autoantibodies identify a novel diisopropylfluorophosphate-binding protein in the lysosomes of normal human neutrophils. J Clin Invest 84: 1577–87.
[6]
Falk RJ, Terrell RS, Charles LA, Jennette JC (1990) Anti-neutrophil cytoplasmic autoantibodies induce neutrophils to degranulate and produce oxygen radicals in vitro. Proc Natl Acad Sci U S A 87: 4115–9.
[7]
Williams JM, Ben-Smith A, Hewins P, Dove SK, Hughes P, et al. (2003) Activation of the G(i) heterotrimeric G protein by ANCA IgG F(ab′)2 fragments is necessary but not sufficient to stimulate the recruitment of those downstream mediators used by intact ANCA IgG. J Am Soc Nephrol 14: 661–9.
[8]
Weidner S, Neupert W, Goppelt-Struebe M, Rupprecht HD (2001) Antineutrophil cytoplasmic antibodies induce human monocytes to produce oxygen radicals in vitro. Arthritis Rheum 44: 1698–706.
[9]
Heeringa P, Brouwer E, Klok PA, Huitema MG, van den Born J, et al. (1996) Autoantibodies to myeloperoxidase aggravate mild anti-glomerular- basement-membrane-mediated glomerular injury in the rat. Am J Pathol 149: 1695–706.
[10]
Little MA, Smyth L, Salama AD, Mukherjee S, Smith J, et al. (2009) Experimental autoimmune vasculitis: an animal model of anti-neutrophil cytoplasmic autoantibody-associated systemic vasculitis. Am J Pathol 174: 1212–20.
[11]
Xiao H, Schreiber A, Heeringa P, Falk RJ, Jennette JC (2007) Alternative Complement Pathway in the Pathogenesis of Disease Mediated by Anti-Neutrophil Cytoplasmic Autoantibodies. Am J Pathol 170: 52–64.
[12]
Huugen D, van Esch A, Xiao H, Peutz-Kootstra CJ, Buurman WA, et al. (2007) Inhibition of complement factor C5 protects against anti-myeloperoxidase antibody-mediated glomerulonephritis in mice. Kidney Int 71: 646–54.
[13]
Xiao H, Heeringa P, Liu Z, Huugen D, Hu P, et al. (2005) The Role of Neutrophils in the Induction of Glomerulonephritis by Anti-Myeloperoxidase Antibodies. Am J Pathol 167: 39–45.
[14]
Xiao H, Heeringa P, Hu P, Liu Z, Zhao M, et al. (2002) Antineutrophil cytoplasmic autoantibodies specific for myeloperoxidase cause glomerulonephritis and vasculitis in mice. J Clin Invest 110: 955–63.
[15]
Bansal PJ, Tobin MC (2004) Neonatal microscopic polyangiitis secondary to transfer of maternal myeloperoxidase-antineutrophil cytoplasmic antibody resulting in neonatal pulmonary hemorrhage and renal involvement. Ann Allergy Asthma Immunol 93: 398–401.
[16]
Pfister H, Ollert M, Froehlich LF, Quintanilla-Martinez L, Colby TV, et al. (2004) Anti-neutrophil cytoplasmic autoantibodies (ANCA) against the murine homolog of proteinase 3 (Wegener's autoantigen) are pathogenic in vivo. Blood 104: 1411–8.
[17]
Rauova L, Gilburd B, Zurgil N, Blank M, Guegas LL, et al. (2002) Induction of biologically active antineutrophil cytoplasmic antibodies by immunization with human apoptotic polymorphonuclear leukocytes. Clin Immunol 103: 69–78.
[18]
van der Geld YM, Hellmark T, Selga D, Heeringa P, Huitema MG, et al. (2007) Rats and mice immunised with chimeric human/mouse proteinase 3 produce autoantibodies to mouse Pr3 and rat granulocytes. Ann Rheum Dis 66: 1679–82.
[19]
Schreiber A, Otto B, Ju X, Zenke M, Goebel U, et al. (2005) Membrane Proteinase 3 Expression in Patients with Wegener's Granulomatosis and in Human Hematopoietic Stem Cell-Derived Neutrophils. J Am Soc Nephrol 16: 2216–24.
[20]
Shultz LD, Lyons BL, Burzenski LM, Gott B, Chen X, et al. (2005) Human lymphoid and myeloid cell development in NOD/LtSz-scid IL2R gamma null mice engrafted with mobilized human hemopoietic stem cells. J Immunol 174: 6477–89.
[21]
Primo VC, Marusic S, Franklin CC, Goldmann WH, Achaval CG, et al. (2010) Anti-PR3 immune responses induce segmental and necrotizing glomerulonephritis. Clin Exp Immunol 159: 327–37.
[22]
Traggiai E, Chicha L, Mazzucchelli L, Bronz L, Piffaretti JC, et al. (2004) Development of a human adaptive immune system in cord blood cell-transplanted mice. Science 304: 104–7.
[23]
Ishikawa F, Yasukawa M, Lyons B, Yoshida S, Miyamoto T, et al. (2005) Development of functional human blood and immune systems in NOD/SCID/IL2 receptor gamma chain(null) mice. Blood 106: 1565–73.
[24]
Kallenberg CG (2010) Pathophysiology of ANCA-associated small vessel vasculitis. Curr Rheumatol Rep 12: 399–405.
[25]
Jennette JC, Falk RJ, Andrassy K, Bacon PA, Churg J, et al. (1994) Nomenclature of systemic vasculitides. Proposal of an international consensus conference. Arthritis Rheum 37: 187–92.
[26]
Williams JM, Pettitt TR, Powell W, Grove J, Savage CO, et al. (2007) Antineutrophil cytoplasm antibody-stimulated neutrophil adhesion depends on diacylglycerol kinase-catalyzed phosphatidic acid formation. J Am Soc Nephrol 18: 1112–20.
[27]
Radford DJ, Lord JM, Savage CO (1999) The activation of the neutrophil respiratory burst by anti-neutrophil cytoplasm autoantibody (ANCA) from patients with systemic vasculitis requires tyrosine kinases and protein kinase C activation. Clin Exp Immunol 118: 171–9.
[28]
Hussain A, Pankhurst T, Goodall M, Colman R, Jefferis R, et al. (2009) Chimeric IgG4 PR3-ANCA induces selective inflammatory responses from neutrophils through engagement of Fcgamma receptors. Immunology 128: 236–44.
[29]
Duffield JS, Tipping PG, Kipari T, Cailhier J-F, Clay S, et al. (2005) Conditional Ablation of Macrophages Halts Progression of Crescentic Glomerulonephritis. Am J Pathol 167: 1207–19.
[30]
Li B, Cohen A, Hudson TE, Motlagh D, Amrani DL, et al. (2010) Mobilized human hematopoietic stem/progenitor cells promote kidney repair after ischemia/reperfusion injury. Circulation 121: 2211–20.
[31]
Lin SL, Castano AP, Nowlin BT, Lupher ML Jr, Duffield JS (2009) Bone marrow Ly6Chigh monocytes are selectively recruited to injured kidney and differentiate into functionally distinct populations. J Immunol 183: 6733–43.
[32]
Park KM, Byun JY, Kramers C, Kim JI, Huang PL, et al. (2003) Inducible nitric-oxide synthase is an important contributor to prolonged protective effects of ischemic preconditioning in the mouse kidney. J Biol Chem 278: 27256–66.
[33]
Taneda S, Hudkins KL, Cui Y, Farr AG, Alpers CE, et al. (2003) Growth factor expression in a murine model of cryoglobulinemia. Kidney Int 63: 576–90.
[34]
Lin SL, Kisseleva T, Brenner DA, Duffield JS (2008) Pericytes and perivascular fibroblasts are the primary source of collagen-producing cells in obstructive fibrosis of the kidney. Am J Pathol 173: 1617–27.
