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A.A. Leonardi, J.S. Curnow, V.L. Calder; Multiple cytokine evaluation in tears of allergic conjunctivitis patients by Multi–Cytokine Bead Assay . Invest. Ophthalmol. Vis. Sci. 2004;45(13):625.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: To measure proinflammatory (Th1–type and Th2–type) cytokine and chemokine levels in tears of patients with different types of allergic conjunctivitis. Methods:Tear samples were obtained from 14 normal subjects (CT), and patients affected by active vernal keratoconjunctivitis (VKC) (N=18), seasonal allergic conjunctivitis (SAC) (N=12), atopic keratoconjunctivitis (AKC) (N=6) and giant papillary conjunctivitis (GPC) (N=3). Concentrations of IL–1ß, –2, –4, –5, –6, –8, –10, –12, –13, IFNγ, TNFα, Eotaxin, RANTES and MCP–1 were determined using multiplexed bead analysis. Samples were incubated with a cocktail of LuminexTM beads, each coated with an individual monoclonal anti–cytokine/chemokine antibody, followed by biotinylated polyclonal antibodies and streptavidin–phycoerythrin. The fluorescence intensity of each bead population was measured using a Luminex100TM. The concentration of each molecule was determined from a standard curve of known concentrations of recombinant molecules. Results: In normal tears, low levels of IL–6, IL–8, MCP–1 and RANTES were found. VKC showed a significant increase over CT of IL–1ß, –2, –4, –5, –6, –10, –12, –13, IFNγ, MCP–1 and eotaxin, SAC of IL–1ß and IL–2, and AKC of IL–1ß, –6, –8 and MCP–1 over CT. In GPC, IL–8 was increased over CT, VKC and SAC, while IL–5 was significantly increased in VKC over all the other groups. When the disease activity score was correlated with cytokine levels significant correlations were found only in SAC, withTh2–type cytokines significantly correlating with each other. The Th1 cytokines, IL–12 and IFNγ, correlated with each other but also with the Th2 cytokines, while IL–2 did not. Eotaxin, MCP–1, RANTES and IL–8 correlated with each other and with both Th1 and Th2 cytokines. TNF was significantly correlated with Th1 and Th2 cytokines, chemokines and the other proinflammatory cytokines IL–1ß, and IL–6. Conclusions: Multiplexed bead analysis is to be a new and helpful method to measure multiple cytokine and chemokine production in tears and to compare the different cytokine patterns in different ocular allergic diseases.
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