Abstract | PURPOSE: The purpose of this study was to determine levels of IL-2, -4, -5, -10, TNF-alpha, and IFN-gamma in aqueous humor (AH) from patients with active panuveitis, anterior uveitis (AU), and noninflammatory controls by using a flow cytometric mutiplex array (CBA) and to compare with results from ELISA. METHODS: Pooled normal AH was spiked with six cytokines at decreasing concentrations for evaluating the CBA. AH was also obtained from 10 controls ( cataract patients) and 36 patients with active uveitis. Cell-free supernatants were added to a cocktail of capture beads and detector antibodies or to antibody-coated wells for CBA and ELISA determination, respectively. RESULTS: CBA demonstrated greater sensitivity for detecting IL-4, IL-10, and TNF-alpha than with ELISA. Increased IFN-gamma was detected in both AU and panuveitis groups compared with controls (P < 0.01). IL-10 was higher in the panuveitis group on steroids (P < 0.01). IL-5 was detected in the control (P < 0.01) and AU groups (P < 0.05) but was undetectable in the panuveitis group (n = 10). Correlations between IFN-gamma and IL-10 were found in all uveitis groups (P < 0.01) but not in controls, whereas TNF-alpha correlations with IL-4/IFN-gamma were obtained in controls but not in the uveitis groups (P < 0.01). CONCLUSIONS: It was possible to measure cytokines titrated into normal AH specimens by CBA, and a greater number of cytokines were detected with increased sensitivity than with ELISA. Elevated IFN-gamma in active uveitis and decreased IL-5 in posterior uveitis suggest Th1 polarity is more marked, with greater uveal tract involvement. The increased IL-10 in the steroid treated group suggests glucocorticoid-induced IL-10 upregulation.
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Authors | Kenneth G-J Ooi, Grazyna Galatowicz, Hamish M A Towler, Susan L Lightman, Virginia L Calder |
Journal | Investigative ophthalmology & visual science
(Invest Ophthalmol Vis Sci)
Vol. 47
Issue 1
Pg. 272-7
(Jan 2006)
ISSN: 0146-0404 [Print] United States |
PMID | 16384973
(Publication Type: Comparative Study, Journal Article, Research Support, Non-U.S. Gov't)
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Chemical References |
- Glucocorticoids
- Interleukin-5
- Interleukin-10
- Interferon-gamma
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Topics |
- Adult
- Aqueous Humor
(metabolism)
- Enzyme-Linked Immunosorbent Assay
(methods)
- Flow Cytometry
(methods)
- Glucocorticoids
(therapeutic use)
- Humans
- Interferon-gamma
(metabolism)
- Interleukin-10
(metabolism)
- Interleukin-5
(metabolism)
- Middle Aged
- Reproducibility of Results
- Sensitivity and Specificity
- Up-Regulation
- Uveitis
(drug therapy, metabolism)
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