Effects of subconjunctival administration of anti-high mobility group box 1 on dry eye in a mouse model of Sjӧgren's syndrome

• Published in: PloS one Vol. 12; no. 8; p. e0183678
• Main Authors: Kim, Kyeong Hwan, Kim, Dong Hyun, Jeong, Hyun Jeong, Ryu, Jin Suk, Kim, Yu Jeong, Oh, Joo Youn, Kim, Mee Kum, Wee, Won Ryang
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 2017; Public Library of Science (PLoS)
• Subjects: Animals; Antibodies; Antigens; Autoimmune diseases; Biology and Life Sciences; Biomedical research; Cell activation; Cell density; Chemokines; Conjunctiva; Conjunctiva - pathology; Cornea; Cytokines; Cytometry; Damage patterns; Dendritic cells; Deoxyribonucleic acid; Disease; Disease Models, Animal; DNA; Dry Eye Syndromes - drug therapy; Enzyme-Linked Immunosorbent Assay; Eye; Flow Cytometry; Glands; HMGB1 protein; HMGB1 Protein - administration & dosage; HMGB1 Protein - immunology; HMGB1 Protein - therapeutic use; Hospitals; Immunology; Inflammation; Interferon; Interleukin 10; Interleukin 17; Interleukin 22; Laboratories; Lymph nodes; Lymphocytes B; Lymphoid cells; Medicine; Medicine and Health Sciences; Mice; Mice, Inbred NOD; Mobility; Pathogenesis; Physical Sciences; Plasma cells; Proteins; Research and Analysis Methods; Sjogren's Syndrome - drug therapy; Staining; T cell receptors; Tearing; Toll-like receptors
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0183678

Extracellular high mobility group box 1 (HMGB1) acts as a damage associated molecular pattern molecule through the Toll-like receptor to promote autoreactive B cell activation, which may be involved in the pathogenesis of Sjӧgren's syndrome. The aim of this study was to investigate the effect of subconjunctival administration of anti-HMGB1 on dry eye in a mouse model of Sjӧgren's syndrome. Ten weeks-old NOD.B10.H2b mice were subconjunctivally injected with 0.02 to 2 μg of anti-HMGB1 antibodies or PBS twice a week for two consecutive weeks. Tear volume and corneal staining scores were measured and compared between before- and after-treatment. Goblet cell density was counted in PAS stained forniceal conjunctiva and inflammatory foci score (>50 cells/focus) was measured in extraorbital glands. Flow cytometry was performed to evaluate the changes in BrdU+ cells, IL-17-, IL-10-, or IFNγ-secreting cells, functional B cells, and IL-22 secreting innate lymphoid cells (ILC3s) in cervical lymph nodes. The level of IL-22 in intraorbital glands was measured by ELISA. Injection of 2 μg or 0.02 μg anti-HMGB1 attenuated corneal epithelial erosions and increased tear secretion (p<0.05). Goblet cell density was increased in 0.2 μg and 2 μg anti-HMGB1-treated-mice with marginal significance. The inflammatory foci score, and the number of BrdU+ cells, IL-17-, IL-10-, IFNγ-secreting cells, and functional B cells did not significantly change following anti-HMGB1 treatment. Surprisingly, the percentage of ILC3s was significantly increased in the draining lymph nodes (p<0.05), and the expression of IL-22 was significantly increased in the intraorbital glands (p<0.05) after administration of 2 μg anti-HMGB1. This study shows that subconjunctival administration of anti-HMGB1 attenuates clinical manifestations of dry eye. The improvement of dry eye may involve an increase of ILC3s, rather than modulation of B or plasma cells, as shown using a mouse model of Sjӧgren's syndrome.