Measurement of plasma cytokines & chemokine’s
Luminex 200 xMAP Technology (Luminex Corp, Austin, TX) was used to measure plasma chemokine’s, C-C motif ligand2 (CCL2), C-C motif ligand4 (CCL4), C-C motif chemokine 11 (CCL11), C-X-C motif chemokine ligand8(CXCL8), C-X-C motif chemokine ligand 10 (CXCL10), interleukin-1 (IL-1), Interleukin-5 (IL-5) and Interleukin-6 (IL-6) in plasma. Briefly, plasma was diluted to two folds, then incubated with specific label beads (Molecular Probes; Life Technologies, Carlsbad, CA) coated with antibodies, after several washes and incubation periods, the previous beads were detected with Luminex system (Luminex Corp, Austin, Tex, USA). Data were obtained and calculated using a 5-parametric curve fit using xPONENT®, version 4.03 in a blinded fashion with measurement performed with the FlexMAP3D system (Luminex Corp, Austin, Tex, USA)[15] .