title: Multimodal analysis of ocular inflammation using the endotoxin-induced uveitis mouse model.
abstract: Endotoxin-induced uveitis (EIU) in rodents is a model of acute Toll-like receptor 4 (TLR4)-mediated organ inflammation, and has been used to model human anterior uveitis, examine leukocyte trafficking and test novel anti-inflammatory therapeutics. Wider adoption has been limited by the requirement for manual, non-specific, cell-count scoring of histological sections from each eye as a measure of disease severity. Here, we describe a comprehensive and efficient technique that uses ocular dissection and multimodal tissue analysis. This allows matched disease scoring by multicolour flow cytometric analysis of the inflammatory infiltrate, protein analysis on ocular supernatants and qPCR on remnant tissues of the same eye. Dynamic changes in cell populations could be identified and mapped to chemokine and cytokine changes over the course of the model. To validate the technique, dose-responsive suppression of leukocyte infiltration by recombinant interleukin-10 was demonstrated, as well as selective suppression of the monocyte (CD11b+Ly6C+) infiltrate, in mice deficient for eitherCcl2orCcr2 Optical coherence tomography (OCT) was used for the first time in this model to allowin vivoimaging of infiltrating vitreous cells, and correlated with CD11b+Ly6G+ counts to provide another unique measure of cell populations in the ocular tissue. Multimodal tissue analysis of EIU is proposed as a new standard to improve and broaden the application of this model.
authors.affiliation: School of Clinical Sciences, University of Bristol, Bristol BS8 1TD, UK colin.chu@bristol.ac.uk a.dick@bristol.ac.uk.
authors.forename: Colin J
authors.initials: CJ
authors.lastname: Chu
authors.affiliation: UCL Institute of Ophthalmology, 11-43 Bath Street, London EC1V 9EL, UK.
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Gardner
authors.affiliation: School of Clinical Sciences, University of Bristol, Bristol BS8 1TD, UK.
authors.forename: David A
authors.initials: DA
authors.lastname: Copland
authors.affiliation: UCL Institute of Ophthalmology, 11-43 Bath Street, London EC1V 9EL, UK.
authors.forename: Sidath E
authors.initials: SE
authors.lastname: Liyanage
authors.affiliation: UCL Institute of Ophthalmology, 11-43 Bath Street, London EC1V 9EL, UK.
authors.forename: Anai
authors.initials: A
authors.lastname: Gonzalez-Cordero
authors.affiliation: UCL Institute of Ophthalmology, 11-43 Bath Street, London EC1V 9EL, UK.
authors.forename: Sophia-Martha
authors.initials: SM
authors.lastname: Kleine Holthaus
authors.affiliation: UCL Institute of Ophthalmology, 11-43 Bath Street, London EC1V 9EL, UK Roche Pharmaceutical Research and Early Development, Ophthalmology Discovery & Biomarkers, Roche Innovation Center Basel, F. Hoffmann-La Roche Ltd, Grenzacherstrasse 124, Basel 4070, Switzerland.
authors.forename: Ulrich F O
authors.initials: UF
authors.lastname: Luhmann
authors.affiliation: UCL Institute of Ophthalmology, 11-43 Bath Street, London EC1V 9EL, UK.
authors.forename: Alexander J
authors.initials: AJ
authors.lastname: Smith
authors.affiliation: UCL Institute of Ophthalmology, 11-43 Bath Street, London EC1V 9EL, UK NIHR Biomedical Research Centre for Ophthalmology at Moorfields Eye Hospital and UCL Institute of Ophthalmology, London EC1V 9EL, UK.
authors.forename: Robin R
authors.initials: RR
authors.lastname: Ali
authors.affiliation: School of Clinical Sciences, University of Bristol, Bristol BS8 1TD, UK UCL Institute of Ophthalmology, 11-43 Bath Street, London EC1V 9EL, UK NIHR Biomedical Research Centre for Ophthalmology at Moorfields Eye Hospital and UCL Institute of Ophthalmology, London EC1V 9EL, UK colin.chu@bristol.ac.uk a.dick@bristol.ac.uk.
authors.forename: Andrew D
authors.initials: AD
authors.lastname: Dick
mesh_terms: D000818:Animals; D002452:Cell Count; D018932:Chemokine CCL2 / Q000378:metabolism; D016207:Cytokines / Q000378:metabolism; D004195:Disease Models, Animal; D004731:Endotoxins; D005123:Eye / Q000473:pathology*; D005260:Female; D005434:Flow Cytometry; D007249:Inflammation / Q000473:pathology*; D008810:Mice, Inbred C57BL; D020556:Neutrophil Infiltration; D009504:Neutrophils / Q000473:pathology; D054390:Receptors, CCR2 / Q000172:deficiency / Q000378:metabolism; D012720:Severity of Illness Index; D041623:Tomography, Optical Coherence; D014605:Uveitis / Q000139:chemically induced* / Q000473:pathology*
chemical_list: C515335:Ccr2 protein, mouse; D018932:Chemokine CCL2; D016207:Cytokines; D004731:Endotoxins; D054390:Receptors, CCR2
keywords: EIU; Flow cytometry; OCT