• 1. Gelatt KN, MacKay EO. Prevalence of the breed-related glaucomas in pure-bred dogs in North America. Vet Ophthalmol 2004; 7:97111.

  • 2. Gelatt KN. The canine glaucomas. In: Gelatt KN, ed. Veterinary ophthalmology. 4th ed. Philadelphia: Wiley-Blackwell, 2007; 753811.

  • 3. Grus F, Sun D. Immunological mechanisms in glaucoma. Semin Immunopathol 2008; 30:121126.

  • 4. Wax MB, Tezel G. Immunoregulation of RGC fate in glaucoma. Exp Eye Res 2009; 88:825830.

  • 5. Shazly TA, Aljajeh M, Latina MA. Autoimmune basis of glaucoma. Semin Ophthalmol 2011; 26:278281.

  • 6. Wax MB. The case for autoimmunity in glaucoma. Exp Eye Res 2011; 93:187190.

  • 7. Hammam T, Montgomery D & Morris D, et al. Prevalence of serum autoantibodies and paraproteins in patients with glaucoma. Eye 2008; 22:349353.

    • Search Google Scholar
    • Export Citation
  • 8. Joachim SC, Bruns K & Lackner KJ, et al. Antibodies to αB-crystallin, vimentin, and heat shock protein 70 in aqueous humor of patients with normal tension glaucoma and IgG antibody patterns against retinal antigen in aqueous humor. Curr Eye Res 2007; 32:501509.

    • Search Google Scholar
    • Export Citation
  • 9. Joachim SC, Pfeiffer N, Grus FH. Autoantibodies in patients with glaucoma: a comparison of IgG serum antibodies against retinal, optic nerve, and optic nerve head antigens. Graefes Arch Clin Exp Ophthalmol 2005; 243:817823.

    • Search Google Scholar
    • Export Citation
  • 10. Joachim SC, Reichelt J & Berneiser S, et al. Sera of glaucoma patients show autoantibodies against myelin basic protein and complex autoantibody profiles against human optic nerve antigens. Graefes Arch Clin Exp Ophthalmol 2008; 246:573580.

    • Search Google Scholar
    • Export Citation
  • 11. Joachim SC, Wuenschig D & Pfeiffer N, et al. IgG antibody patterns in aqueous humor of patients with primary open angle glaucoma and pseudoexfoliation glaucoma. Mol Vis 2007; 13:15731579.

    • Search Google Scholar
    • Export Citation
  • 12. Reichelt J, Joachim SC & Pfeiffer N, et al. Analysis of autoantibodies against human retinal antigens in sera of patients with glaucoma and ocular hypertension. Curr Eye Res 2008; 33:253261.

    • Search Google Scholar
    • Export Citation
  • 13. Tezel G, Edward DP, Wax MB. Serum autoantibodies to optic nerve head glycosaminoglycans in patients with glaucoma. Arch Ophthalmol 1999; 117:917924.

    • Search Google Scholar
    • Export Citation
  • 14. Boehm N, Wolters D & Thiel U, et al. New insights into autoantibody profiles from immune-privileged sites in the eye: a glaucoma study. Brain Behav Immun 2012; 26:96102.

    • Search Google Scholar
    • Export Citation
  • 15. Dervan EW, Chen H & Ho SL, et al. Protein macroarray profiling of serum autoantibody profiles in pseudoexfoliation glaucoma. Investig Ophth Vis Sci 2010; 51:29682975.

    • Search Google Scholar
    • Export Citation
  • 16. Singer HS, Loiselle CR & Lee O, et al. Anti-basal ganglia antibody abnormalities in Sydenham chorea. J Neuroimmunol 2003; 136:154161.

  • 17. Wendlandt JT, Grus FH & Hansen BH, et al. Striatal antibodies in children with Tourette's syndrome: multivariate discriminate analysis of IgG repertoires. J Neuroimmunol 2001; 119:106113.

    • Search Google Scholar
    • Export Citation
  • 18. Sharshar T, Lacroix-Desmazes S & Mouthon L, et al. Selective impairment of serum antibody repertoires toward muscle and thymus antigens in patients with seronegative and seropositive myasthenia gravis. Eur J Immunol 1998; 28:23442354.

    • Search Google Scholar
    • Export Citation
  • 19. Zephir H, Leblanc D & Dubucquoi S, et al. Serum IgG repertoire in clinically isolated syndrome predicts multiple sclerosis. Mult Scler 2009; 15:593600.

    • Search Google Scholar
    • Export Citation
  • 20. Cekaite L, Hovig E, Sioud M. Protein arrays: a versatile toolbox for target identification and monitoring of patient immune responses. Methods Mol Biol 2007; 360:335348.

    • Search Google Scholar
    • Export Citation
  • 21. Chanseaud Y, Tamby MC & Guilpain P, et al. Analysis of autoantibody repertoires in small- and medium-sized vessels vasculitides. Evidence for specific perturbations in polyarteritis nodosa, microscopic polyangiitis, Churg-Strauss syndrome and Wegener's granulomatosis. J Autoimmun 2005; 24:169179.

    • Search Google Scholar
    • Export Citation
  • 22. Caligiuri G, Stahl D & Kaveri S, et al. Autoreactive antibody repertoire is perturbed in atherosclerotic patients. Lab Invest 2003; 83:939947.

    • Search Google Scholar
    • Export Citation
  • 23. Calkins DJ. Critical pathogenic events underlying progression of neurodegeneration in glaucoma. Prog Retin Eye Res 2012; 31:702719.

    • Search Google Scholar
    • Export Citation
  • 24. Nickells RW. Ganglion cell death in glaucoma: from mice to men. Vet Ophthal 2007; 10:8894.

  • 25. Pascale A, Drago F, Govoni S. Protecting the retinal neurons from glaucoma: lowering ocular pressure is not enough. Pharmacol Res 2012; 66:1932.

    • Search Google Scholar
    • Export Citation
  • 26. Libby RT, Li Y & Savinova OV, et al. Susceptibility to neurodegeneration in a glaucoma is modified by Bax gene dosage. PLoS Genet 2005; 1:e4.

    • Search Google Scholar
    • Export Citation
  • 27. Iwabe S, Moreno-Mendoza NA & Trigo-Tavera F, et al. Retrograde axonal transport obstruction of brain-derived neurotrophic factor (BDNF) and its TrkB receptor in the retina and optic nerve of American Cocker Spaniels with spontaneous glaucoma. Vet Ophthal 2007; 10:1219.

    • Search Google Scholar
    • Export Citation
  • 28. Ekesten B, Narfstrom K. Correlation of morphologic features of the iridocorneal angle to intraocular pressures in Samoyeds. Am J Vet Res 1991; 52:18751878.

    • Search Google Scholar
    • Export Citation
  • 29. Lowry DH, Rosenberg NJ & Farr AL, et al. Protein measurement with folin phenol reagent. J Biol Chem 1951; 193:265275.

  • 30. Giuliano EA, Moore CP. Diseases and surgery of the lacrimal secretory system. In: Gelatt KN, ed. Veterinary ophthalmology. 4th ed. Philadelphia: Wiley-Blackwell, 2007; 633661.

    • Search Google Scholar
    • Export Citation
  • 31. Shields MB. Normal tension glaucoma: is it different from primary open angle glaucoma? Curr Opin Ophthalmol 2008; 19:8588.

  • 32. Gordon MO, Beiser JA & Brandt JD, et al. The Ocular Hypertension Treatment Study: baseline factors that predict the onset of primary open-angle glaucoma. Arch Ophthalmol 2002; 120:714720.

    • Search Google Scholar
    • Export Citation
  • 33. Reilly CM, Morris R, Dubielzig RR. Canine goniodysgenesis-related glaucoma: a morphologic review of 100 cases looking at inflammation and pigment dispersion. Vet Ophthalmol 2005; 8:253258.

    • Search Google Scholar
    • Export Citation
  • 34. Mangan BG, Al-yahya K & Chen C-T, et al. Retinal pigment epithelial damage, breakdown of the blood-retinal barrier, and retinal inflammation in dogs with primary glaucoma. Vet Ophthalmol 2007; 10:117124.

    • Search Google Scholar
    • Export Citation
  • 35. Jiang B, Harper MM & Kecova H, et al. Neuroinflammation in advanced canine glaucoma. Mol Vis 2010; 16:20922108.

  • 36. Zhou X, Li F & Kong L, et al. Involvement of inflammation, degradation, and apoptosis in a mouse model of glaucoma. J Biol Chem 2005; 280:3124031248.

    • Search Google Scholar
    • Export Citation
  • 37. Anderson MG, Nair KS & Amonoo LA, et al. GpnmbR150X allele must be present in bone marrow derived cells to mediate DBA/2J glaucoma. BMC Genetics 2008; 9:3044.

    • Search Google Scholar
    • Export Citation
  • 38. Ripoll VM, Irvine KM & Ravasi T, et al. Gpnmb is induced in macrophages by IFN-gamma and lipopolysaccharide and acts as a feedback regulator of proinflammatory responses. J Immunol 2007; 178:65576566.

    • Search Google Scholar
    • Export Citation
  • 39. Joachim SC, Grus FH & Kraft D, et al. Complex antibody profile changes in an experimental autoimmune glaucoma animal model. Invest Ophthalmol Vis Sci 2009; 50:48224827.

    • Search Google Scholar
    • Export Citation
  • 40. Joachim SC, Wax MB & Seidel P, et al. Enhanced characterization of serum autoantibody reactivity following HSP 60 immunization in a rat model of experimental autoimmune glaucoma. Exp Eye Res 2010; 35:900908.

    • Search Google Scholar
    • Export Citation
  • 41. Bakalash S, Kipnis J & Yoles E, et al. Resistance of retinal ganglion cells to an increase in intraocular pressure is immune-dependent. Invest Ophthalmol Vis Sci 2002; 43:26482653.

    • Search Google Scholar
    • Export Citation
  • 42. Tezel G, Hernandez MR, Wax MB. Immunostaining of heat shock proteins in the retina and optic nerve head of normal and glaucomatous eyes. Arch Ophthalmol 2000; 118:511518.

    • Search Google Scholar
    • Export Citation
  • 43. Tezel G, Seigel GM, Wax MB. Autoantibodies to small heat shock proteins in glaucoma. Invest Ophthalmol Vis Sci 1998; 39:22772287.

  • 44. Wax MB, Tezel G & Kawase K, et al. Serum autoantibodies to heat shock proteins in glaucoma patients from Japan and the United States. Ophthalmology 2001; 108:296302.

    • Search Google Scholar
    • Export Citation
  • 45. Wax MB, Tezel G & Yang J, et al. Induced autoimmunity to heat shock proteins elicits glaucomatous loss of retinal ganglion cell neurons via activated T-cell–derived Fas-ligand. J Neurosci 2008; 28:1208512096.

    • Search Google Scholar
    • Export Citation
  • 46. Yu AL, Fuchshofer R & Birke M, et al. Oxidative stress and TGF-β2 increase heat shock protein 27 expression in human optic nerve head astrocytes. Invest Ophthalmol Vis Sci 2008; 49:54035411.

    • Search Google Scholar
    • Export Citation
  • 47. Yang J, Patil RV & Gordon M, et al. T cell subsets and sIL-2R/IL-2 levels in patients with glaucoma. Am J Ophthalmol 2001; 131:421426.

  • 48. Rönkkö S, Rekonen P & Kaarniranta K, et al. Phospholipase A2 in chamber angle of normal eyes and patients with primary open angle glaucoma and exfoliation glaucoma. Mol Vis 2007; 13:408417.

    • Search Google Scholar
    • Export Citation
  • 49. Grus FH, Joachim SC & Bruns K, et al. Serum autoantibodies to α-fodrin are present in glaucoma patients from Germany and the United States. Invest Ophthalmol Vis Sci 2006; 47:968976.

    • Search Google Scholar
    • Export Citation
  • 50. Kremmer S, Kreuzfelder E & Klein R, et al. Antiphosphaditylserine antibodies are elevated in normal tension glaucoma. Clin Exp Immunol 2001; 125:211215.

    • Search Google Scholar
    • Export Citation
  • 51. Maruyama I, Ohguro H, Ikeda Y. Retinal ganglion cells recognized by serum autoantibody against γ-enolase found in glaucoma patients. Invest Ophthalmol Vis Sci 2000; 41:16571665.

    • Search Google Scholar
    • Export Citation
  • 52. Romano C, Barrett DA & Li Z, et al. Anti-rhodopsin antibodies in sera from patients with normal-pressure glaucoma. Invest Ophthalmol Vis Sci 1995; 36:19681975.

    • Search Google Scholar
    • Export Citation
  • 53. Lee KJ, Jeong SM & Hoehn BD, et al. Valosin-containing protein is a novel autoantigen in patients with glaucoma. Optom Vis Sci 2011; 88:164172.

    • Search Google Scholar
    • Export Citation
  • 54. Maruyama I, Maeda T & Okisaka S, et al. Autoantibody against neuron-specific enolase found in glaucoma patients causes retinal dysfunction in vivo. Jpn J Ophthalmol 2002; 46:112.

    • Search Google Scholar
    • Export Citation
  • 55. Berger T, Rubner P & Schautzer F, et al. Antimyelin antibodies as a predictor of clinically definite multiple sclerosis after a first demyelinating event. N Engl J Med 2003; 349:139145.

    • Search Google Scholar
    • Export Citation
  • 56. Shmerling RH. Autoantibodies in systemic lupus erythematosus—there before you know it. N Engl J Med 2003; 349:14991500.

  • 57. Arbuckle MR, McClain MT & Rubertone MV, et al. Development of autoantibodies before the clinical onset of systemic lupus erythematosus. N Engl J Med 2003; 349:15261533.

    • Search Google Scholar
    • Export Citation
  • 58. Tezel G, Wax MB. Glaucoma. Chem Immunol Allergy 2007; 92:221227.

  • 59. Poletaev A, Osipenko L. General network of natural autoantibodies as immunological homunculus (immunculus). Autoimmun Rev 2003; 2:264271.

    • Search Google Scholar
    • Export Citation
  • 60. Gilmour MA, Cardenas MR & Blaik MA, et al. Evaluation of a comparative pathogenesis between cancer-associated retinopathy in humans and sudden acquired retinal degeneration syndrome in dogs via diagnostic imaging and western blot analysis. Am J Vet Res 2006; 67:877881.

    • Search Google Scholar
    • Export Citation
  • 61. Keller RL, Kania SA & Hendrix DV, et al. Evaluation of canine serum for the presence of antiretinal autoantibodies in sudden acquired retinal degeneration syndrome. Vet Ophthalmol 2006; 9:195200.

    • Search Google Scholar
    • Export Citation
  • 62. Braus BK, Hauck SM & Ammann B, et al. Neuron-specific enolase antibodies in patients with sudden acquired retinal degeneration syndrome. Vet Immunol Immunopathol 2008; 124:177183.

    • Search Google Scholar
    • Export Citation

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Western blot patterns of serum autoantibodies against optic nerve antigens in dogs with goniodysgenesis-related glaucoma

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  • 1 Department of Clinical Sciences, Cummings School of Veterinary Medicine, Tufts University, North Grafton, MA 01536.
  • | 2 Department of Clinical Sciences, Cummings School of Veterinary Medicine, Tufts University, North Grafton, MA 01536.
  • | 3 Department of Clinical Sciences, Cummings School of Veterinary Medicine, Tufts University, North Grafton, MA 01536.
  • | 4 Department of Biomedical Sciences, Cummings School of Veterinary Medicine, Tufts University, North Grafton, MA 01536.
  • | 5 Biostatistics Research Center, Institute for Clinical Research and Health Policy Studies, Tufts Medical Center, 800 Washington St, Boston, MA 02111.

Abstract

Objective—To investigate whether differences existed between clinically normal dogs and dogs with goniodysgenesis-related glaucoma (GDRG) in serum autoantibodies against optic nerve antigens.

Animals—16 dogs with GDRG, 17 healthy dogs with unremarkable pectinate ligament and iridocorneal angle morphology, and 13 euthanized dogs with no major ocular abnormalities or underlying diseases.

Procedures—Western blotting was performed with optic nerve extracts from the euthanized dogs as an antigen source and serum from clinically normal dogs and dogs with GDRG as a primary antibody (autoantibody) source. Blots were evaluated for presence and density of bands.

Results—Multiple bands were identified on western blots from all dogs with GDRG and all clinically normal dogs, with a high degree of variability among individual dogs. Dogs with GDRG were significantly more likely than healthy dogs to have bands present at 38, 40, and 68 kDa. Dogs with GDRG had significant increases in autoreactivity at 40 and 53 kDa and a significant decrease in autoreactivity at 48 kDa.

Conclusions and Clinical Relevance—Significant differences in serum autoantibodies against optic nerve antigens were found in dogs with versus without GDRG. Although it remains unclear whether these differences were part of the pathogenesis of disease or were sequelae to glaucomatous changes, these findings provide support for the hypothesis that immune-mediated mechanisms play a role in the development or progression of GDRG. However, the high degree of variability among individual dogs and the considerable overlap between groups suggest that the clinical usefulness of this technique for distinguishing dogs with GDRG from clinically normal dogs is likely limited.

Abstract

Objective—To investigate whether differences existed between clinically normal dogs and dogs with goniodysgenesis-related glaucoma (GDRG) in serum autoantibodies against optic nerve antigens.

Animals—16 dogs with GDRG, 17 healthy dogs with unremarkable pectinate ligament and iridocorneal angle morphology, and 13 euthanized dogs with no major ocular abnormalities or underlying diseases.

Procedures—Western blotting was performed with optic nerve extracts from the euthanized dogs as an antigen source and serum from clinically normal dogs and dogs with GDRG as a primary antibody (autoantibody) source. Blots were evaluated for presence and density of bands.

Results—Multiple bands were identified on western blots from all dogs with GDRG and all clinically normal dogs, with a high degree of variability among individual dogs. Dogs with GDRG were significantly more likely than healthy dogs to have bands present at 38, 40, and 68 kDa. Dogs with GDRG had significant increases in autoreactivity at 40 and 53 kDa and a significant decrease in autoreactivity at 48 kDa.

Conclusions and Clinical Relevance—Significant differences in serum autoantibodies against optic nerve antigens were found in dogs with versus without GDRG. Although it remains unclear whether these differences were part of the pathogenesis of disease or were sequelae to glaucomatous changes, these findings provide support for the hypothesis that immune-mediated mechanisms play a role in the development or progression of GDRG. However, the high degree of variability among individual dogs and the considerable overlap between groups suggest that the clinical usefulness of this technique for distinguishing dogs with GDRG from clinically normal dogs is likely limited.

Contributor Notes

Dr. Pumphrey's present address is Southern New Hampshire Veterinary Referral Hospital, 336 Abby Rd, Manchester, NH 03103.

Supported by the Tufts University Cummings School of Veterinary Medicine Fund for Companion Animal Health.

Presented in abstract form at the 47th Annual Meeting of the American College of Veterinary Ophthalmologists, Hilton Head, SC, October 2011.

The authors thank Holly Jameson for technical assistance.

Address correspondence to Dr. Pizzirani (stefano.pizzirani@tufts.edu).