VALUE = Relative expression level compared to the average six internal controls
Standard deviation =
According to the manufacturer's instructions (R & D).
Samples were lyophilized and diluted again in a protein buffer for homogenization
Unstimulated whole saliva was collected by direct spitting into 50ml Falcon tubes placed on ice. Supernatant was obtained following centrifugation of whole saliva for 15 minutes at 2600 g.
Samples were mixed with with C5a, IL-4, IL-32 alpha, CD40 ligand, IL-5, CXCL10/IP-10, G-CSF, IL-6, CXCL11/I-TAC, GM-CSF, IL-8, CCL2/MCP-1, CXCL1/GRO alpha, IL-10, MIF, CCL1/I-309, IL-12 p70, CCL3/MIP-1 alpha, ICAM-1, IL-13, CCL4/MIP-1 beta, IFN-gamma, IL-16, CCL5/RANTES, IL-1 alpha, IL-17, CXCL12/SDF-1, IL-1 beta, IL-17E, Serpin E1/PAI-1, IL-1ra, IL-23, TNF-alpha, IL-2 and IL-27 and the hybridized with capture antibodies spotted in duplicate on nitrocellulose membranes. After over night incubation and 3 washing steps, Streptavidin-Horseradish Peroxidase and chemiluminescent detection reagents are added and the signal produced is in proportion to the amount of cytokine bound.
Raw data were processed with the Optiquant Array Computational Tool (Packard Bioscience) for background correction and normalization. Data analysis and statistical evaluations were performed with SAM alogrthm (http://www-stat.stanford.edu/~tibs/SAM/sam.pdf)
Array scanning was performed according to the manufacturer's instructions (R & D).