Mcp 1

The saliva samples were then thawed and total protein saliva was quantified using the Qubit protein Assay Kit (Thermofisher Scientific). Salivary IgA (Human IgA Platinum ELISA, ebioscience, Affymetrix, Santa Clare, CA), lactoferrin, lysozyme (AssayPro, St. Charles, MO, USA), Groα, Groβ and MCP-1 (Elabscience, Houston, Texas) were measured by ELISA according to the manufacturer’s instructions. The limit of detection for IgA was: 1.6 ng/ml; lactoferrin: 0.625 ng/ml; lysozyme: 0.0781 ng/ml; Groα: 15.63 pg/ml, Groβ: 15.63 pg/ml and MCP-1: 15.63 pg/ml. Data were expressed as the concentration of each molecule relative to total saliva protein concentration [13 (link)]. Those determinations of relative salivary IgA concentration with a ratio higher than 1 were excluded of the study due to possible contaminations [14 (link)]. All determinations were performed in triplicate.

Saliva samples were thawed and total saliva protein was quantified using Qubit protein Assay Kit (molecular probes Life Technologies). Salivary IgA (Human IgA, Platinum ELISA, ebioscience, Affymetrix, Santa Clare, CA), Lactoferrin, Lysozyme (AssayPro, St, Charles, MO, USA), Gro-alpha/CXCL1, Gro-beta/CXCL2 and MCP-1 (Elabscience, Houston, Texas) were measured by ELISA according to manufacturer’s instructions. Limits of detection were for IgA: 1.6 ng/ml; lactoferrin: 0.625 ng/ml; Lysozyme: 0.0781 ng/ml; Gro-alpha/CXCL1: 15.63 pg/ml, Gro-beta/CXCL2: 15.63 pg/ml, MCP-1: 15.63 pg/ml, Angiogenin: 1.64 pg/ml, ARCP30: 24.69 pg/ml and Siglec 5: 6.86 pg/ml. The data was expressed as concentration of each protein relative to total saliva protein concentration.

The serum cytokine and chemokine levels were measured using a mouse ELISA kit, according to the manufacturer’s instructions (IL-4 and IL-6, Enzo Life Sciences, Farmingdale, NY, USA; TSLP, MCP-1, Elabscience, Huston, ID, USA; IL-1β, TARC, MDC, and RANTES, R&D Systems Inc., Minneapolis, MN, USA).