• Histology of different tissues (e.g. retina, optic nerve, brain) using light and fluorescence microscopy
• Ophthalmologic Examinations (e.g. IOP measurement, fundus examination, OCT)
• Intravitreal injection of therapeutic compounds
• Proteomics using mass spectrometry 
• Immunoproteomics using Western blotting (1D and 2D) und antigen microarray

 

During the last years an autoimmune involvement has been discussed in the pathogenesis of glaucoma. In several studies we could show significant differences in antibody profiles of glaucoma patients in comparison to healthy subjects. These antibody patterns provide hints for changes in the autoimmunity in glaucoma patients, but this does not necessarily mean that glaucoma is an autoimmune disease.

Are these antibodies an epiphenomenon or are some of them causative at least in a subset of glaucoma patients?

In order to test the hypothesis that autoimmunity can be involved in the pathogenesis of glaucoma, we are developed an animal model of Experimental Autoimmune Glaucoma (EAG).

Similar to the procedures used in autoimmune uveitis (EAU) or experimental autoimmune encephalitis (EAE), we immunize the animals with possible antigens (previously identified in glaucoma patients) and then investigate possible subsequent ganglion cell loss and antibody binding.

 

 

We are analyzing the immunological modifications of the complex antibody profiles in the nerve crush animal model. Histological analyses of the retina and optic nerve will be performed to find out if retinal ganglion cell loss occurs in these animals.

Studies suggest that beneficial T cell-dependent immunity is a physiological response to CNS trauma partially counteracting the trauma-induced damage during nerve crush. But there is no knowledge about the modifications of the antibody profiles after optic nerve crush.

Because of the small blood and aqueous humor sample volumes in animals we developed a method to detect antibody-antigen-reactivities through Protein G beads and ProteinChip technology.

 

Immunological modifications of the complex antibody profiles in a ischemia reperfusion model are also currently examined. And compared to findings in the Experimental Autoimmune Glaucoma Model.

 

Acetylcholine regulates perfusion of numerous organs via changes in local blood flow involving muscarinic receptor-induced release of nitric oxide. Five muscarinic acetylcholine receptor subtypes, denoted M1 through M5, have been identified. In ocular arteries, expression and functional relevance of muscarinic receptors remains unknown at present. The purpose of our research project is to determine the expression pattern of muscarinic receptor subtypes in retinal and ciliar arteries in humans and mice. Another major focus of our project is to investigate the impact of muscarinic receptors on vascular responses using in vitro ocular vascular preparations.

These projects are supported in part by Boehringer Ingelheim Stiftung, Grimmke Stiftung, MAIFOR (Universitätsmedizin), Gertrud Kusen-Stiftung, and others.

 

• Hu J, Sisignano M, Brecht R, Perumal N, Angioni C, Bibli IS, Fisslthaler B, Kleinert H, Pfeiffer N, Fleming I, Manicam C (2021) Cyp2c44 epoxygenase-derived epoxyeicosatrienoic acids in vascular smooth muscle cells elicit vasoconstriction of the murine ophthalmic artery. Sci Rep 11:18764.
• Herzog DP, Perumal N, Manicam C, Treccani G, Nadig J, Rossmanith M, Engelmann J, Jene T, Hasch A, van der Kooij MA, Lieb K, Gassen NC, Grus FH, Muller MB (2021) Longitudinal CSF proteome profiling in mice to uncover the acute and sustained mechanisms of action of rapid acting antidepressant (2R,6R)-hydroxynorketamine (HNK). Neurobiol Stress 15:100404.
• Birk M, Baum E, Zadeh JK, Manicam C, Pfeiffer N, Patzak A, Helmstadter J, Steven S, Kuntic M, Daiber A, Gericke A (2021) Angiotensin II Induces Oxidative Stress and Endothelial Dysfunction in Mouse Ophthalmic Arteries via Involvement of AT1 Receptors and NOX2. Antioxidants (Basel) 10.
• Liu H, Perumal N, Manicam C, Mercieca K, Prokosch V (2020) Proteomics Reveals the Potential Protective Mechanism of Hydrogen Sulfide on Retinal Ganglion Cells in an Ischemia/Reperfusion Injury Animal Model. Pharmaceuticals (Basel) 13.
• Teister J, Liu A, Wolters D, Pfeiffer N, Grus FH (2018) Peripapillary fluorescence lifetime reveals age-dependent changes using fluorescence lifetime imaging ophthalmoscopy in rats. Exp Eye Res 176:110-120.
• Teister J, Anders F, Beck S, Funke S, von Pein H, Prokosch V, Pfeiffer N, Grus F (2017) Decelerated neurodegeneration after intravitreal injection of alpha-synuclein antibodies in a glaucoma animal model. Sci Rep 7:6260.
• Gramlich OW, Teister J, Neumann M, Tao X, Beck S, von Pein HD, Pfeiffer N, Grus FH (2016) Immune response after intermittent minimally invasive intraocular pressure elevations in an experimental animal model of glaucoma. J Neuroinflammation 13:82.
• Gramlich OW, Lorenz K, Grus FH, Kriechbaum M, Ehrlich D, Humpel C, Fischer-Colbrie R, Bechrakis NE, Troger J (2014a) Catestatin-like immunoreactivity in the rat eye. Neuropeptides 48:7-13.
• Gramlich OW, Lueckner TC, Kriechbaum M, Teister J, Tao X, von Pein HD, Pfeiffer N, Grus FH (2014b) Dynamics, alterations, and consequences of minimally invasive intraocular pressure elevation in rats. Invest Ophthalmol Vis Sci 55:600-611.
• Gramlich OW, Beck S, von Thun Und Hohenstein-Blaul N, Boehm N, Ziegler A, Vetter JM, Pfeiffer N, Grus FH (2013) Enhanced insight into the autoimmune component of glaucoma: IgG autoantibody accumulation and pro-inflammatory conditions in human glaucomatous retina. PLoS One 8:e57557.
• Gramlich O, Beck S, Ziegler A, Pfeiffer N, Grus F (2013) Autoimmune component in glaucoma: IgG autoantibody accumulation, plasma cells and microglia under pro-inflammatory conditions. Invest Ophthalmol Vis Sci 54:3176-.
• Teister J, Gramlich O, Lueckner T, Kriechbaum M, Pfeiffer N, Grus F (2013) Investigation of the retinal thickness using Spectralis OCT(R) reveals a significant decrease after short time elevation of IOP. Invest Ophthalmol Vis Sci 54:1458-.
• Lueckner T, Gramlich O, Kriechbaum M, Teister J, Pfeiffer N, Grus F (2013) Suction-cup oculopression offers minimal-invasive opportunities of arbitrary IOP elevations in rats. Invest Ophthalmol Vis Sci 54:1985-.
• Gericke A, Goloborodko E, Sniatecki JJ, Steege A, Wojnowski L, Pfeiffer N (2013) Contribution of nitric oxide synthase isoforms to cholinergic vasodilation in murine retinal arterioles. Exp Eye Res 109:60-66.
• Gramlich OW, von Pein HD, Ziegler A, Bitz K, Pfeiffer N, Grus FH (2012) Topical Treatment With A Selective COX-2 Inhibitor Promotes Retinal Ganglion Cell Survival After Optic Nerve Crush. Invest Ophthalmol Vis Sci 53:6273-.
• Laspas P, Gramlich OW, Muller HD, Cuny CS, Gottschling PF, Pfeiffer N, Dick HB, Joachim SC, Grus FH (2011) Autoreactive antibodies and loss of retinal ganglion cells in rats induced by immunization with ocular antigens. Invest Ophthalmol Vis Sci 52:8835-8848.
• Gramlich OW, Joachim SC, Gottschling PF, Laspas P, Cuny CS, Pfeiffer N, Grus FH (2011) Ophthalmopathology in rats with MBP-induced experimental autoimmune encephalomyelitis. Graefes Arch Clin Exp Ophthalmol 249:1009-1020.