Multiple Natural and Experimental Inflammatory Rabbit Lacrimal Gland Phenotypes

Austin K Mircheff; Yanru Wang; Joel E Schechter; Meng Li; Warren Tong; Mayssa Attar; Murty Chengalvala; Joe Harmuth; Jeffery J Prusakiewicz

doi:10.1016/j.jtos.2016.07.001

Multiple Natural and Experimental Inflammatory Rabbit Lacrimal Gland Phenotypes

Ocul Surf. 2016 Oct;14(4):460-483.e3. doi: 10.1016/j.jtos.2016.07.001. Epub 2016 Jul 15.

Authors

Austin K Mircheff¹, Yanru Wang², Joel E Schechter³, Meng Li⁴, Warren Tong⁵, Mayssa Attar⁵, Murty Chengalvala⁶, Joe Harmuth⁶, Jeffery J Prusakiewicz⁷

Affiliations

¹ Department of Physiology & Biophysics, Keck School of Medicine, University of Southern California, Los Angeles, CA, USA; Department of Ophthalmology, Keck School of Medicine, University of Southern California, Los Angeles, CA, USA. Electronic address: amirchef@usc.edu.
² Department of Physiology & Biophysics, Keck School of Medicine, University of Southern California, Los Angeles, CA, USA.
³ Department of Cell & Neurobiology, Keck School of Medicine, University of Southern California, Los Angeles, CA, USA.
⁴ Bioinformatics Service, Norris Medical Library, University of Southern California, Los Angeles, CA, USA.
⁵ Translational Drug Metabolism, Pharmacokinetics and Immunology, Allergan Inc., Irvine, CA, USA.
⁶ Immunology Services, Covance Research Products, Denver, PA, USA.
⁷ Drug Metabolism, Covance Laboratories Inc., Madison, WI, USA.

Abstract

Purpose: To investigate lacrimal gland (LG) immunophysiological and immune-mediated inflammatory process (IMIP) phenotype diversity.

Methods: Ex vivo matured dendritic cells (mDC) were loaded with acinar cell microparticles (M_P). Peripheral blood lymphocytes (PBL) were activated in mixed cell reactions with mDC and injected directly into autologous, unilateral LG (1°ATD-LG) of two rabbit cohorts, one naïve, one immunized with a LG lysate membrane fraction (P_i). Autoimmune IgG titers were assayed by ELISA, MCR PBL stimulation indices (SI) by [³H]-thymidine incorporation. Schirmer tests without and with topical anesthetic (STT-I, STT-I_A) and rose Bengal (RB) staining tests were performed. H&E and immunohistochemically stained sections were examined. RNA yields and selected transcript abundances were measured. Immune cell number and transcript abundance data were submitted to Principal Component Analysis (PCA).

Results: Immunizing P_i dose influenced SI but not IgG titers. STT scores were decreased, and rose Bengal scores increased, by day 118 after immunization. Previous immunization exacerbated scores in 1°ATD-eyes and exacerbated 1°ATD-LG atrophy. IMIP were evident in 2°ATD-LG as well as 1°ATD-LG. PCA described diverse immunophysiological phenotypes in control LG and diverse IMIP phenotypes in ATD-LG. IgG titers and SI pre-adoptive transfer were significantly associated with certain post-adoptive transfer IMIP phenotype features, and certain LG IMIP features were significantly associated with RB and STT I_A scores.

Conclusions: The underlying variability of normal states may contribute to the diversity of experimental IMIP phenotypes. The ability to generate and characterize diverse phenotypes may lead to phenotype-specific diagnostic and therapeutic paradigms.

Keywords: Sjögren disease; acinar cells; autoantigens; dacryoadenitis; dendritic cells; dry eye; keratoconjunctivitis.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Adoptive Transfer
Animals
Autoantibodies / blood
Autoantigens / immunology*
Autoimmunity
Blotting, Western
Dacryocystitis / immunology*
Dendritic Cells / immunology
Disease Models, Animal*
Enzyme-Linked Immunosorbent Assay
Female
Immunoglobulin G / blood
Immunophenotyping
Lacrimal Apparatus / immunology*
Lymphocyte Activation
Principal Component Analysis
Rabbits
Real-Time Polymerase Chain Reaction
Rose Bengal

Substances

Autoantibodies
Autoantigens
Immunoglobulin G
Rose Bengal

Grants and funding

P30 DK048522/DK/NIDDK NIH HHS/United States