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Richardson Foundation","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100001315","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100001209","name":"Knights Templar Eye Foundation","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100001209","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Med."],"abstract":"<jats:sec>\n                    <jats:title>Purpose<\/jats:title>\n                    <jats:p>To investigate IL-17 related mechanisms for developing dry eye disease in the Pinkie mouse strain with a loss of function RXR\u03b1 mutation.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>Measures of dry eye disease were assessed in the cornea and conjunctiva. Expression profiling was performed by single-cell RNA sequencing (scRNA-seq) to compare gene expression in conjunctival immune cells. Conjunctival immune cells were immunophenotyped by flow cytometry and confocal microscopy. The activity of RXR\u03b1 ligand 9-cis retinoic acid (RA) was evaluated in cultured monocytes and \u03b3\u03b4 T cells.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      Compared to wild type (WT) C57BL\/6, Pinkie has increased signs of dry eye disease, including decreased tear volume, corneal barrier disruption, corneal\/conjunctival cornification and goblet cell loss, and corneal vascularization, opacification, and ulceration with aging. ScRNA-seq of conjunctival immune cells identified \u03b3\u03b4 T cells as the predominant IL-17 expressing population in both strains and there is a 4-fold increased percentage of \u03b3\u03b4 T cells in Pinkie. Compared to WT, IL-17a, and IL-17f significantly increased in Pinkie with conventional T cells and \u03b3\u03b4 T cells as the major producers. Flow cytometry revealed an increased number of IL-17\n                      <jats:sup>+<\/jats:sup>\n                      \u03b3\u03b4 T cells in Pinkie. Tear concentration of the IL-17 inducer IL-23 is significantly higher in Pinkie. 9-cis RA treatment suppresses stimulated IL-17 production by \u03b3\u03b4 T and stimulatory activity of monocyte supernatant on \u03b3\u03b4 T cell IL-17 production. Compared to WT bone marrow chimeras, Pinkie chimeras have increased IL-17\n                      <jats:sup>+<\/jats:sup>\n                      \u03b3\u03b4 T cells in the conjunctiva after desiccating stress and anti-IL-17 treatment suppresses dry eye induced corneal MMP-9 production\/activity and conjunctival goblet cell loss.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>These findings indicate that RXR\u03b1 suppresses generation of dry eye disease-inducing IL-17 producing lymphocytes s in the conjunctiva and identifies RXR\u03b1 as a potential therapeutic target in dry eye.<\/jats:p>\n                  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