Simultaneous quantification of 17 immune mediators in aqueous humour from patients with corneal rejection

Purpose: To simultaneously quantitate and compare the concentrations of 17 immune mediators: (1) the cytokines interleukin-1 beta, IL-2, IL-4, IL-5, IL-6, IL-7, IL-10, IL-12p70, IL-13, IL-17, tumour necrosis factor-alpha, interferon-gamma; (2) the growth factors granulocyte-monocyte colony-stimulating factor (GM-CSF) and granulocyte colony-stimulating factor (G-CSF), and (3) the chemokines CXCL-8, monocyte chemoattractant protein-1, and macrophage inflammatory protein-1 beta in aqueous humour from patients with corneal rejection and patients with a non-inflammatory condition in the anterior chamber. Methods: Aqueous humour was obtained by paracentesis of the anterior chamber in 14 patients with corneal rejection, three patients with cataract and six patients with Fuchs' endothelial dystrophy. Simultaneous quantitation of 17 mediators in 25 mu l aqueous humour from each patient was performed by employing a highly sensitive Luminex (R) 100 multiplex array assay. Results: All 17 immune mediators were detected in aqueous humour from rejection patients. The ranges of the immune mediators were determined. The immune mediators were signifcantly increased in aqueous humour from rejection patients compared with that from other patients. Conclusions: The Luminex 100 multiplex array assay is very efficient in simultaneous quantitation of multiple immune mediators in small volumes of aqueous humour. A total of 17 immune mediators were increased in aqueous humour from rejection patients. This underlines the complex immunological interactions of the rejection process.