Multiplex bead immunoassay analysis of aqueous humor reveals distinct cytokine profiles in uveitis.
OBJECTIVE
To extensively characterize the complex network of cytokines present in uveitis aqueous humor (AqH), and the relationships between cytokines and the cellular infiltrate.


METHODS
AqH from noninflammatory control subjects and patients with idiopathic, Fuchs' heterochromic cyclitis (FHC), and herpes-viral or Behçet's uveitis were analyzed for IL-1beta, -2, -4, -5, -7, -8, -10, -12, -13, -15, TNFalpha, IFNgamma, CCL2 (MCP-1), CCL5 (RANTES), CCL11 (Eotaxin), TGFbeta2, and CXCL12 (SDF-1), using multiplex bead immunoassays. The cellular infiltrate was also determined for each sample.


RESULTS
Idiopathic uveitis AqH, compared with noninflammatory controls, was characterized by high levels of IL-6, IL-8, CCL2 and IFNgamma, the levels of which correlated with each other. For IL-6 and IL-8 these levels were proportional to the number of neutrophils present. By contrast, the levels of both TGFbeta2 and CXCL12 decreased in idiopathic uveitis AqH with increasing inflammation. Cluster analysis showed a degree of segregation between noninflammatory and idiopathic uveitis AqH. Further examination using random forest analysis yielded a complete distinction between these two groups. The minimum cytokines required for this classification were IL-6, IL-8, CCL2, IL-13, TNFalpha, and IL-2.


CONCLUSIONS
Application of multiplex bead immunoassays has allowed us to identify distinct patterns of cytokines that relate to both clinical disease and the cellular infiltrates present. Bioinformatics analysis allowed identification of cytokines that differentiate idiopathic uveitis from noninflammatory control AqH and are likely to be important for the pathogenesis of uveitis.
Curnow
S John
SJ
Institute of Biomedical Research, MRC Centre for Immune Regulation, Division of Immunity and Infection, Medical School, The University of Birmingham, UK. s.j.curnow@bham.ac.uk
Falciani
Francesco
F
Durrani
Omar M
OM
Cheung
C M Gemmy
CM
Ross
Emma J
EJ
Wloka
Kaska
K
Rauz
Saaeha
S
Wallace
Graham R
GR
Salmon
Mike
M
Murray
Philip I
PI
D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001082:Aqueous Humor / Q000378:metabolism*; D015415:Biomarkers / Q000378:metabolism; D016000:Cluster Analysis; D016207:Cytokines / Q000378:metabolism*; D005136:Eye Proteins / Q000378:metabolism*; D005260:Female; D006801:Humans; D007118:Immunoassay / Q000379:methods; D008297:Male; D008863:Microspheres; D008875:Middle Aged; D015864:Panuveitis / Q000145:classification / Q000378:metabolism*; D014606:Uveitis, Anterior / Q000145:classification / Q000378:metabolism*
D015415:Biomarkers; D016207:Cytokines; D005136:Eye Proteins