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Background/Objectives Uveitis is a leading cause of blindness globally. Common aetiologies include infectious or immune-mediated causes, but diagnosis can be challenging and up to a third of patients remain ‘idiopathic’. We sought to identify clinical and immunological biomarkers at acute presentation which facilitate diagnosis.Methods We prospectively recruited acute uveitis patients from two tertiary centres over 18 months. Careful clinical, serological, visual, and radiological phenotyping was performed. 37 cytokines/chemokines/growth factors were evaluated in patients, and 50 age/sex-matched controls using a Milliplex® Panel or ELISA.Results 67 patients (37 female, median age 42 (range 16–79) years), were recruited with a median duration of follow-up of 18.5 months (range 12–37 months). 17, 29 and 21 were classified as infectious, immune-mediated and idiopathic, respectively. A relapsing course was more common in the immune-mediated (20/29, 69%) and idiopathic (12/21, 57%) groups compared to the infectious group (3/17, 18%; p=0.0005). 95 affected eyes demonstrated worse visual outcomes in the infectious (median logmar 0.21, IQR 0–0.31) and idiopathic (0.16; 0.02–0.33) groups compared to the immune-mediated group (0; 0–0.19; p=0.05). Macular thickness at nadir was higher in the immune-mediated (270 µm; 260–293) and infectious groups (269; 250–303) compared to the idiopathic group (250; 223–277; p=0.037). Preliminary cytokine/chemokine analysis showed several elevated cytokines/chemokines in uveitis patients compared to healthy controls and different cytokine/chemokine signatures in immune-mediated, infectious and idiopathic aetiologies.Conclusions/Discussion Ongoing analysis will provide a clinical and cytokine/chemokine signature which may assist with improved diagnostic profiling of patients with acute uveitis, to guide management and improve outcomes.