Cytokines, matrix metalloproteases, angiogenic and growth factors in tears of normal subjects and vernal keratoconjunctivitis patients.
BACKGROUND
To detect the presence of multiple mediators and growth factors in tears of vernal keratoconjunctivitis (VKC) patients with active disease using stationary phase antibody arrays.


METHODS
Tears were collected from 12 normal subjects (CT) and 24 active VKC patients. Tears were centrifuged and successively probed using three microwell plate arrays specific for: (i) cytokines: interleukin (IL)-2, IL-4, IL-5, IL-8, IL-10, IL-12, IL-13, interferon-gamma and tumour necrosis factor-alpha; (ii) growth factors: basic fibroblast growth factor (bFGF), platelet-derived growth factor, thrombopoietin, angiopoietin-2, vascular endothelial growth factor (VEGF), hepatocyte growth factor (HGF), keratocyte growth factor, tissue inhibitor of metalloprotease (TIMP)-1 and heparin-binding epithelial growth factor (HB-EGF) and (iii) matrix metalloprotease (MMP)-1, MMP-2, MMP-3, MMP-8, MMP-9, MMP-10, MMP-13, TIMP-1 and TIMP-2.


RESULTS
Interleukin-8 signals were detected in all CT and highly detected in all VKC samples. The Th2-type cytokines, IL-4, IL-5 and IL-10 were detected only in tears of VKC patients. Signals for bFGF, HB-EGF, VEGF and HGF were detected in 41-87% of VKC samples and in few CT samples. Only TIMP-1 and TIMP-2 were found in all normal and patient tear samples, whereas MMP-1, MMP-2, MMP-3, MMP-9 and MMP-10 were highly present in all VKC samples.


CONCLUSIONS
Stationary phase antibody array methodology was useful for the screening of various cytokines, growth factors and MMPs in tears. These analyses identified in tears of VKC patients previously unreported factors including MMP-3 and MMP-10 and multiple proteases, growth factors and cytokines, which may all play an important role in the pathogenesis of conjunctival inflammation.
Leonardi
A
A
Department of Neuroscience, Ophthalmology Unit, University of Padua, Padua, Italy.
Sathe
S
S
Bortolotti
M
M
Beaton
A
A
Sack
R
R
D000293:Adolescent; D000328:Adult; D043925:Angiogenesis Inducing Agents / Q000032:analysis / Q000276:immunology*; D002648:Child; D002675:Child, Preschool; D003233:Conjunctivitis, Allergic / Q000276:immunology* / Q000378:metabolism; D016207:Cytokines / Q000032:analysis / Q000276:immunology*; D005260:Female; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000032:analysis / Q000276:immunology*; D008297:Male; D045726:Metalloproteases / Q000032:analysis / Q000276:immunology*; D040081:Protein Array Analysis; D013666:Tears / Q000737:chemistry / Q000201:enzymology / Q000276:immunology*; D055815:Young Adult
D043925:Angiogenesis Inducing Agents; D016207:Cytokines; D036341:Intercellular Signaling Peptides and Proteins; D045726:Metalloproteases