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A.A. Leonardi, V. Calder, J.S. Curnow, S. Sathe, R. Sack; Metalloproteases, Cytokines and Growth Factors Detection in Tears of Vernal Keratoconjunctivitis (VKC) Patients by Antibody Array Analysis . Invest. Ophthalmol. Vis. Sci. 2006;47(13):5871.
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© ARVO (1962-2015); The Authors (2016-present)
To detect the presence and distribution of multiple mediators and growth factors in tears of active vernal keratoconjunctivitis (VKC) patients using stationary phase antibody array (MA).
Tears were collected from 6 normal subjects and 16 active VKC patients. Tears were centrifuged and successively probed using three microwell plate arrays specific for: a) MMP–1, –2, –3, –8, –9, –10, –13, TIMP–1 and –2; b) angiogenic factors FGF–b, PDGF, TPO, ANG–2, FGFb, VEGF and EGFhb c) cytokines: IL–1, –3, –4, –5, –6, –7, –8, –10, –12, –13, IFNγ and TNFα. Samples were also probed with a membrane array optimized to detect a larger number of inflammatory mediators and chemokines.
Only TIMP–1 and –2 were found at high levels in normal tears. In VKC, MMPs 1, 2, 3, 9, and 10 were detected, with very high levels of MMP–3, –9 and –10. Tarsal–VKC samples demonstrated a higher expression of MMPs compared to limbal–VKC. 12/16 VKC samples contained high levels of FGFb and EGF–hb, while 2/6 of the controls were also positive. Th2–cytokines and pro–inflammatory cytokines were highly expressed only in VKC patients.
Stationary phase antibody arrays are a useful tool for the screening of a large number of mediators in low volumes of tears. Analysis allowed the identification of previously undetected factors in tears of VKC patients (i.e., MMP–3 and MMP–10) and confirmed the presence of a high expression of multiple MMPs, growth factors and cytokines that contribute to the pathogenesis of conjunctival allergic inflammation.
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