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Janet L. Davis, Anat Galor, Philip Ruiz, Alexandra Amador, Thomas Albini; Simultaneous Cytokine and Cytofluorographic Analysis of Aqueous Humor in Active Uveitis. Invest. Ophthalmol. Vis. Sci. 2011;52(14):6561.
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To create immunologic profiles of patients with active uveitis using simultaneous cytokine and cytofluorographic determinations in small volume specimens of aqueous humor.
Patients with active uveitis consented to anterior chamber paracentesis to obtain specimens of aqueous humor and phlebotomy to obtain blood for research purposes. 0.1 to 0.2 ml of aqueous humor was acquired with a 30-32 g needle under topical anesthesia in the clinic or under surgical anesthesia in the operating room and transferred immediately to the Transplantation Laboratory. The specimen was centrifuged and the supernatant submitted for a 17 cytokine Luminex assay. The cell pellet was resuspended and examined with cytofluorography to determine cell type. Cytokine analysis was performed on serum and cytofluorography on the white blood cell buffy coat. Cytofluorography was gated for both lymphocyte and myelomonocyte sized cells.
There were 10 patients, 8 female and 2 male with age range from 5 to 55, median 25.5 years. Five had anterior uveitis (2 JIA, 2 HLA-B27+ AAU, 1 Fuchs), 3 had intermediate uveitis, and 2 had panuveitis associated with systemic disease (Crohns, Behcet). IL-6 was elevated in most ocular specimens. Patients receiving TNF alpha inhibitors showed evidence of suppression. IL 17 was found in some specimens. CD20 lymphocytes were elevated in two patients receiving long-term immunosuppression.
Small volume specimens of aqueous humor can be used to create immunologic profiles with information about both infiltrating cell types and the cytokine milieu. Ability to resample patients easily with medication changes or changes in inflammatory scores has the potential to enable therapy tailored to the patient's immunologic status.
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