Unraveling the equine lymphocyte proteome: differential septin 7 expression associates with immune cells in equine recurrent uveitis.
Abstract: Equine recurrent uveitis is a spontaneous, lymphocyte-driven autoimmune disease. It affects horses worldwide and presents with painful remitting-relapsing inflammatory attacks of inner eye structures eventually leading to blindness. Since lymphocytes are the key players in equine recurrent uveitis, we were interested in potential changes of their protein repertoire which may be involved in disease pathogenesis. To create a reference for differential proteome analysis, we first unraveled the equine lymphocyte proteome by two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis and subsequently identified 352 protein spots. Next, we compared lymphocytes from ERU cases and healthy horses with a two-dimensional fluorescence difference in gel electrophoresis approach. With this technique, we identified seven differentially expressed proteins between conditions. One of the significantly lower expressed candidates, septin 7, plays a role in regulation of cell shape, motility and migration. Further analyses revealed T cells as the main cell type with decreased septin 7 abundance in equine recurrent uveitis. These findings point to a possible pathogenetic role of septin 7 in this sight-threatening disease.
Publication Date: 2014-03-10 PubMed ID: 24614191PubMed Central: PMC3951111DOI: 10.1371/journal.pone.0091684Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.
The research explores the differences in protein expression within the immune cells of horses suffering from equine recurrent uveitis (ERU), an autoimmune disease that can lead to blindness, and healthy horses. In particular, the study found that the protein septin 7, which influences cell shape, movement and migration, was less prevalent in the immune cells of horses with ERU.
Study Purpose and Methods
- The primary aim of this research was to ascertain the changes that occur in the protein repertoire of lymphocytes, which are crucial players in ERU. This was pursued with the hope of better understanding the pathogenesis, or the biological mechanism leading to the disease development, of ERU.
- The researchers first established a reference for differential proteome analysis by revealing the equine lymphocyte proteome, using a technique known as two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and identifying 352 protein spots.
- Having identified the baseline proteome, the scientists then compared the lymphocytes from ERU-afflicted horses and healthy horses using another method called two-dimensional fluorescence difference in gel electrophoresis.
Findings
- Seven proteins were found to have different levels of expression between the ERU and healthy cases. The examination of these proteins could give insights into the alteration of the immune response in ERU.
- Specifically, one of the proteins called septin 7, was found significantly less in ERU cases. Septin 7 is important in regulating a cell’s shape, its ability to move, and its migration. Lower levels of septin 7 were discovered mainly within the T cells of the immune system.
Implications
- The researchers suggest that the lower abundance of septin 7 may play a key role in the development of ERU.
- This new information could contribute significantly to the understanding of the molecular changes that occur in ERU, paving the way for the development of new treatments or preventative measures for this sight-threatening disease.
Cite This Article
APA
Degroote RL, Hauck SM, Amann B, Hirmer S, Ueffing M, Deeg CA.
(2014).
Unraveling the equine lymphocyte proteome: differential septin 7 expression associates with immune cells in equine recurrent uveitis.
PLoS One, 9(3), e91684.
https://doi.org/10.1371/journal.pone.0091684 Publication
Researcher Affiliations
- Institute of Animal Physiology, Department of Veterinary Sciences, Ludwig Maximilians University Munich, Munich, Germany.
- Research Unit Protein Sciences, Helmholtz Center Munich, German Research Center for Environmental Health, Neuherberg, Germany.
- Institute of Animal Physiology, Department of Veterinary Sciences, Ludwig Maximilians University Munich, Munich, Germany.
- Institute of Animal Physiology, Department of Veterinary Sciences, Ludwig Maximilians University Munich, Munich, Germany.
- Research Unit Protein Sciences, Helmholtz Center Munich, German Research Center for Environmental Health, Neuherberg, Germany; Center for Ophthalmology, Institute for Ophthalmic Research, Eberhard Karls University of Tübingen, Tübingen, Germany.
- Institute of Animal Physiology, Department of Veterinary Sciences, Ludwig Maximilians University Munich, Munich, Germany.
MeSH Terms
- Animals
- Blotting, Western
- Electrophoresis, Gel, Two-Dimensional
- Flow Cytometry
- Horse Diseases / immunology
- Horses / immunology
- Lymphocyte Subsets / metabolism
- Mass Spectrometry
- Proteome / metabolism
- Recurrence
- Septins / metabolism
- T-Lymphocytes / metabolism
- Uveitis / immunology
- Uveitis / veterinary
Conflict of Interest Statement
Competing Interests: The authors have declared that no competing interests exist.
References
This article includes 60 references
- Gilger BC, Michau TM. Equine recurrent uveitis: new methods of management. Vet Clin North Am Equine Pract 20: 417–427, vii.
- Deeg CA, Reese S, Gerhards H, Wildner G, Kaspers B. The uveitogenic potential of retinal S-antigen in horses. Invest Ophthalmol Vis Sci 45: 2286–2292.
- Deeg CA, Ehrenhofer M, Thurau SR, Reese S, Wildner G. Immunopathology of recurrent uveitis in spontaneously diseased horses. Exp Eye Res 75: 127–133.
- Gilger BC, Malok E, Cutter KV, Stewart T, Horohov DW. Characterization of T-lymphocytes in the anterior uvea of eyes with chronic equine recurrent uveitis. Vet Immunol Immunopathol 71: 17–28.
- Schwink KL. Equine uveitis. Vet Clin North Am Equine Pract 8: 557–574.
- Deeg CA, Hauck SM, Amann B, Pompetzki D, Altmann F. Equine recurrent uveitis-a spontaneous horse model of uveitis. Ophthalmic Res 40: 151–153.
- Deeg CA, Thurau SR, Gerhards H, Ehrenhofer M, Wildner G. Uveitis in horses induced by interphotoreceptor retinoid-binding protein is similar to the spontaneous disease. Eur J Immunol 32: 2598–2606.
- Deeg CA, Pompetzki D, Raith AJ, Hauck SM, Amann B. Identification and functional validation of novel autoantigens in equine uveitis. Mol Cell Proteomics 5: 1462–1470.
- Deeg CA, Raith AJ, Amann B, Crabb JW, Thurau SR. CRALBP is a highly prevalent autoantigen for human autoimmune uveitis. Clin Dev Immunol 2007: 39245.
- Deeg CA, Amann B, Raith AJ, Kaspers B. Inter- and intramolecular epitope spreading in equine recurrent uveitis. Invest Ophthalmol Vis Sci 47: 652–656.
- Hauck SM, Dietter J, Kramer RL, Hofmaier F, Zipplies JK. Deciphering membrane-associated molecular processes in target tissue of autoimmune uveitis by label-free quantitative mass spectrometry. Mol Cell Proteomics 9: 2292–2305.
- Keller A, Nesvizhskii AI, Kolker E, Aebersold R. Empirical statistical model to estimate the accuracy of peptide identifications made by MS/MS and database search. Anal Chem 74: 5383–5392.
- Tooley AJ, Gilden J, Jacobelli J, Beemiller P, Trimble WS. Amoeboid T lymphocytes require the septin cytoskeleton for cortical integrity and persistent motility. Nat Cell Biol 11: 17–26.
- Spiess BM. Equine recurrent uveitis: the European viewpoint. Equine Vet J Suppl 50–56.
- Fruhauf B, Ohnesorge B, Deegen E, Boeve M. Surgical management of equine recurrent uveitis with single port pars plana vitrectomy. Vet Ophthalmol 1: 137–151.
- Gilger BC, Salmon JH, Wilkie DA, Cruysberg LP, Kim J. A novel bioerodible deep scleral lamellar cyclosporine implant for uveitis. Invest Ophthalmol Vis Sci 47: 2596–2605.
- LeHoang P, Sterkers M, Thillaye B, de Kozak Y, Coscas G. Primate model of uveoretinitis and vasculitis/experimental autoimmune uveoretinitis induced in cynomolgus monkeys by retinal s antigen. Ophthalmic Res 40: 181–188.
- Wildner G, Diedrichs-Mohring M, Thurau SR. Rat models of autoimmune uveitis. Ophthalmic Res 40: 141–144.
- de Smet MD, Bitar G, Roberge FG, Gery I, Nussenblatt RB. Human S-antigen: presence of multiple immunogenic and immunopathogenic sites in the Lewis rat. J Autoimmun 6: 587–599.
- Caspi RR. Ocular autoimmunity: the price of privilege?. Immunol Rev 213: 23–35.
- Vergara D, Chiriaco F, Acierno R, Maffia M. Proteomic map of peripheral blood mononuclear cells. Proteomics 8: 2045–2051.
- Ramirez-Boo M, Garrido JJ, Ogueta S, Calvete JJ, Gomez-Diaz C. Analysis of porcine peripheral blood mononuclear cells proteome by 2-DE and MS: analytical and biological variability in the protein expression level and protein identification. Proteomics 6 Suppl 1 S215–225.
- Lenchik NI, Desiderio DM, Gerling IC. Two-dimensional gel electrophoresis characterization of the mouse leukocyte proteome, using a tri-reagent for protein extraction. Proteomics 5: 2202–2209.
- Haudek VJ, Slany A, Gundacker NC, Wimmer H, Drach J. Proteome maps of the main human peripheral blood constituents. J Proteome Res 8: 3834–3843.
- Chae JI, Kim J, Lee SG, Koh MW, Jeon YJ. Quantitative proteomic analysis of pregnancy-related proteins from peripheral blood mononuclear cells during pregnancy in pigs. Anim Reprod Sci 134: 164–176.
- Schulz M, Dotzlaw H, Mikkat S, Eggert M, Neeck G. Proteomic analysis of peripheral blood mononuclear cells: selective protein processing observed in patients with rheumatoid arthritis. J Proteome Res 6: 3752–3759.
- Gerling IC, Singh S, Lenchik NI, Marshall DR, Wu J. New data analysis and mining approaches identify unique proteome and transcriptome markers of susceptibility to autoimmune diabetes. Mol Cell Proteomics 5: 293–305.
- Degroote RL, Hauck SM, Kremmer E, Amann B, Ueffing M. Altered expression of talin 1 in peripheral immune cells points to a significant role of the innate immune system in spontaneous autoimmune uveitis. J Proteomics 75: 4536–4544.
- Degroote RL, Hauck SM, Treutlein G, Amann B, Frohlich KJ. Expression Changes and Novel Interaction Partners of Talin 1 in Effector Cells of Autoimmune Uveitis. J Proteome Res .
- Tatton WG, Chalmers-Redman RM, Elstner M, Leesch W, Jagodzinski FB. Glyceraldehyde-3-phosphate dehydrogenase in neurodegeneration and apoptosis signaling. J Neural Transm Suppl 77–100.
- Carlton JG, Martin-Serrano J. Parallels between cytokinesis and retroviral budding: a role for the ESCRT machinery. Science 316: 1908–1912.
- Hashizume S, Kuroda K, Murakami H. Identification of lactoferrin as an essential growth factor for human lymphocytic cell lines in serum-free medium. Biochim Biophys Acta 763: 377–382.
- Mazurier J, Legrand D, Hu WL, Montreuil J, Spik G. Expression of human lactotransferrin receptors in phytohemagglutinin-stimulated human peripheral blood lymphocytes. Isolation of the receptors by antiligand-affinity chromatography. Eur J Biochem 179: 481–487.
- Fischer R, Debbabi H, Dubarry M, Boyaka P, Tome D. Regulation of physiological and pathological Th1 and Th2 responses by lactoferrin. Biochem Cell Biol 84: 303–311.
- Pao LI, Badour K, Siminovitch KA, Neel BG. Nonreceptor protein-tyrosine phosphatases in immune cell signaling. Annu Rev Immunol 25: 473–523.
- Eriksen KW, Woetmann A, Skov L, Krejsgaard T, Bovin LF. Deficient SOCS3 and SHP-1 expression in psoriatic T cells. J Invest Dermatol 130: 1590–1597.
- Sampson MJ, Lovell RS, Craigen WJ. The murine voltage-dependent anion channel gene family. Conserved structure and function. J Biol Chem 272: 18966–18973.
- Maldonado EN, Lemasters JJ. Warburg revisited: regulation of mitochondrial metabolism by voltage-dependent anion channels in cancer cells. J Pharmacol Exp Ther 342: 637–641.
- Okunuki Y, Usui Y, Kezuka T, Hattori T, Masuko K. Proteomic surveillance of retinal autoantigens in endogenous uveitis: implication of esterase D and brain-type creatine kinase as novel autoantigens. Mol Vis 14: 1094–1104.
- Bonilha VL. Focus on molecules: ezrin. Exp Eye Res 84: 613–614.
- Pore D, Parameswaran N, Matsui K, Stone MB, Saotome I. Ezrin Tunes the Magnitude of Humoral Immunity. J Immunol .
- Weirich CS, Erzberger JP, Barral Y. The septin family of GTPases: architecture and dynamics. Nat Rev Mol Cell Biol 9: 478–489.
- Hartwell LH. Genetic control of the cell division cycle in yeast. IV. Genes controlling bud emergence and cytokinesis. Exp Cell Res 69: 265–276.
- Byers B, Goetsch L. A highly ordered ring of membrane-associated filaments in budding yeast. J Cell Biol 69: 717–721.
- Li S, Ou XH, Wei L, Wang ZB, Zhang QH. Septin 7 is required for orderly meiosis in mouse oocytes. Cell Cycle 11: 3211–3218.
- Kremer BE, Adang LA, Macara IG. Septins regulate actin organization and cell-cycle arrest through nuclear accumulation of NCK mediated by SOCS7. Cell 130: 837–850.
- Kremer BE, Haystead T, Macara IG. Mammalian septins regulate microtubule stability through interaction with the microtubule-binding protein MAP4. Mol Biol Cell 16: 4648–4659.
- Kinoshita M, Field CM, Coughlin ML, Straight AF, Mitchison TJ. Self- and actin-templated assembly of Mammalian septins. Dev Cell 3: 791–802.
- Santos J, Cerveira N, Bizarro S, Ribeiro FR, Correia C. Expression pattern of the septin gene family in acute myeloid leukemias with and without MLL-SEPT fusion genes. Leuk Res .
- Engidawork E, Gulesserian T, Fountoulakis M, Lubec G. Aberrant protein expression in cerebral cortex of fetus with Down syndrome. Neuroscience 122: 145–154.
- Jia ZF, Huang Q, Kang CS, Yang WD, Wang GX. Overexpression of septin 7 suppresses glioma cell growth. J Neurooncol 98: 329–340.
- Deeg CA, Kaspers B, Gerhards H, Thurau SR, Wollanke B. Immune responses to retinal autoantigens and peptides in equine recurrent uveitis. Invest Ophthalmol Vis Sci 42: 393–398.
- Caspi R. Autoimmunity in the immune privileged eye: pathogenic and regulatory T cells. Immunol Res 42: 41–50.
- Zipplies JK, Hauck SM, Schoeffmann S, Amann B, van der Meijden CH. Kininogen in autoimmune uveitis: decrease in peripheral blood stream versus increase in target tissue. Invest Ophthalmol Vis Sci 51: 375–382.
- Wasik AA, Polianskyte-Prause Z, Dong MQ, Shaw AS, Yates JR 3rd. Septin 7 forms a complex with CD2AP and nephrin and regulates glucose transporter trafficking. Mol Biol Cell 23: 3370–3379.
- Marko AJ, Miller RA, Kelman A, Frauwirth KA. Induction of glucose metabolism in stimulated T lymphocytes is regulated by mitogen-activated protein kinase signaling. PLoS One 5: e15425.
- Finlay DK. Regulation of glucose metabolism in T cells: new insight into the role of Phosphoinositide 3-kinases. Front Immunol 3: 247.
- Santoni V, Molloy M, Rabilloud T. Membrane proteins and proteomics: un amour impossible?. Electrophoresis 21: 1054–1070.
- Lilley KS, Friedman DB. All about DIGE: quantification technology for differential-display 2D-gel proteomics. Expert Rev Proteomics 1: 401–409.
- Nagaraj N, Lu A, Mann M, Wisniewski JR. Detergent-based but gel-free method allows identification of several hundred membrane proteins in single LC-MS runs. J Proteome Res 7: 5028–5032.
Use Nutrition Calculator
Check if your horse's diet meets their nutrition requirements with our easy-to-use tool Check your horse's diet with our easy-to-use tool
Talk to a Nutritionist
Discuss your horse's feeding plan with our experts over a free phone consultation Discuss your horse's diet over a phone consultation
Submit Diet Evaluation
Get a customized feeding plan for your horse formulated by our equine nutritionists Get a custom feeding plan formulated by our nutritionists