title: Autologous lacrimal-lymphoid mixed-cell reactions induce dacryoadenitis in rabbits.
abstract: Autoimmune dacryoadenitis, such as occurs in Sjögren's syndrome, is a frequent cause of lacrimal insufficiency, which in turn can cause dry eye. Rabbits are used frequently to test ocular therapies. Our goal is to develop a rabbit model of autoimmune dacryoadenitis to identify and test candidate therapies. Our approach arises from the observations that lacrimal gland epithelial cells stimulate proliferation in cultured autologous lymphocyte preparations and that an anti-MHC II antibody blocks this proliferation. The purpose of this study was to determine if injecting this proliferating autologous mixed cell reaction could induce dacryoadenitis in rabbits. After establishing that irradiated lacrimal gland epithelial cells stimulate proliferation in autologous peripheral blood lymphocytes, irradiated cells from a single lacrimal gland were co-cultured with autologous lymphocytes and after 5 days the mixed cell reaction, or components of the reaction, were injected into the contralateral lacrimal gland of the donor rabbit. After 2 weeks, the injected glands were removed and lymphocytic infiltration quantitated using digital image analysis of immunostained histological sections. Injecting an autologous mixed cell reaction of co-cultured irradiated lacrimal gland epithelial cells and lymphocytes reliably induced abundant periductal foci of >200 lymphocytes expressing CD18 and/or a rabbit thymic lymphocyte antigen (RTLA). Injection of medium or autologous lymphocytes alone elicited little response; injections of lymphocytes cultured with lysates of lacrimal gland epithelial cells elicited variable, modest responses. These lysates did not stimulate proliferation in the mixed cell reaction and proliferation was not observed if a porous membrane separated co-cultured lacrimal gland cells and lymphocytes. The results demonstrate that injecting an autologous mixed cell reaction of lacrimal gland epithelial cells and lymphocytes reliably creates a model of autoimmune dacryoadenitis. The relative ineffectiveness of components of the reaction to do the same supports the hypothesis that lacrimal gland epithelial cells trigger or exacerbate lacrimal autoimmune disease by presentation of autoantigens via MHC II. This experimental system can aid efforts to further understand mechanisms of diseases, and to identify and test candidate therapies.
authors.affiliation: Department of Physiology & Biophysics, University of Southern California School of Medicine, Los Angeles, CA, 90033, USA.
authors.forename: Z
authors.initials: Z
authors.lastname: Guo
authors.forename: D
authors.initials: D
authors.lastname: Song
authors.forename: A M
authors.initials: AM
authors.lastname: Azzarolo
authors.forename: J E
authors.initials: JE
authors.lastname: Schechter
authors.forename: D W
authors.initials: DW
authors.lastname: Warren
authors.forename: R L
authors.initials: RL
authors.lastname: Wood
authors.forename: A K
authors.initials: AK
authors.lastname: Mircheff
authors.forename: H R
authors.initials: HR
authors.lastname: Kaslow
mesh_terms: D000818:Animals; D002455:Cell Division / Q000276:immunology; D003607:Dacryocystitis / Q000209:etiology* / Q000276:immunology; D004847:Epithelial Cells / Q000276:immunology; D007765:Lacrimal Apparatus / Q000166:cytology / Q000276:immunology*; D007959:Lymphocyte Culture Test, Mixed; D008214:Lymphocytes / Q000276:immunology*; D008285:Major Histocompatibility Complex / Q000276:immunology; D008297:Male; D011817:Rabbits; D012859:Sjogren's Syndrome / Q000276:immunology*