Novel murine model for autoantibody-induced dry eye disease

Abstract

Purpose : Our previous studies have demonstrated the presence of anti-citrullinated protein autoantibodies (ACPA) in ocular surface washes from patients with autoimmune dry eye disease (DED). Moreover, this pathology was responsive to ocular surface immunoglobulin G (OSIG) treatment. The purpose of this study was to develop a murine model of autoantibody-induced DED to support future mechanistic and pharmacological studies.

Methods : Male C57BL/6J mice (8-10 weeks of age) received four daily instillations (5 µl) of either anti-histone 4 (citrulline R3) antibody (H4R3 ACPA; 100 ng/ml) or anti-histone 4 wildtype antibody (H4 WT; 100ng/ml). Corneal pathology was evaluated at baseline, day 5 and day 8 by fluorescein staining using a hand-held slit lamp with built-in cobalt blue light. Fluorescence was captured using a Spectralis HRT with anterior segment module (Heidelberg Engineering) in fluorescein angiography mode. Fluorescein staining was graded using both the Oxford scoring system and National Eye Institute (NEI) grading system. Data were analyzed by Two-Way ANOVA with Holm-Šídák’s multiple comparisons test.

Results : Eyes treated topically with H4R3 ACPA showed significant corneal damage on day 5 (median score: 2.5, range 2 – 4, n = 6, P < 0.01) and on day 8 (median score: 3.5, range 2 – 4, n = 6, P < 0.01), when compared with baseline. In contrast, eyes treated with H4 WT antibody showed some signs of corneal damage (median score: 1.5, range: 1 – 2, P < 0.01), but pathology was significantly lower than in H4R3 ACPA-treated eyes.

Conclusions : Topical instillation of H4R3 ACPA antiserum results in significant corneal damage in mice mimicking autoimmune DED pathology in patients. The novel model for autoantibody-induced dry eye disease offers a validated and standardized experimental paradigm that will allow for the rigorous testing of targeted therapeutics for autoimmune DED.

This abstract was presented at the 2022 ARVO Annual Meeting, held in Denver, CO, May 1-4, 2022, and virtually.