Laboratory Measurements

We selected to measure 12 proteins (CRP, SAA, MMP1, MMP2, MMP9, sgp130, sICAM1, sVCAM1, sIL2Rα, sIL6R, sTNFR1, and sTNFR2) in our PAGODA study subjects; the selection was based on published literature on role of these proteins in inflammation and MA. Luminex assays for these 12 proteins were obtained from Millipore Inc., Billerica, MA, USA. Multiplex assays were performed according to the instructions provided with the kit. Briefly, serum samples were incubated with antibody coated microspheres, followed by biotinylated detection antibody. Detection of the proteins was accomplished by incubation with phycoerythrin-labeled streptavidin. The resultant bead immuno-complexes were then read on a FLEXMAP3D (Luminex, TX, USA) with the instrument settings recommended by the manufacturer.