Elisa assays

T cell from donors' peripheral blood was primed with dendritic cells pulsed by antigens acquired from breast cancer cells in the presence of interleukins 7 (IL-7, 1 ng/mL, Invitrogen) and 2 (IL-2, 10 u/mL, Gibco) for seven days as previously reported [37 (link)]. The related cytokines were analyzed by related ELISA assays (Sigma, USA).

The levels of various biomarkers in serum, including total cholesterol (TC), high-density lipoprotein cholesterol (HDL), triacylglycerol (TG), glucose, and total protein (TP) were analyzed using a Vitros 250 chemistry system from Ortho Clinical Diagnostics (Ortho Clinical Diagnostics, Raritan, NJ, USA). To calculate the levels of low-density lipoprotein cholesterol (LDL) and very LDL (VLDL), the Friedewald’s formula was used [18 (link)], where LDL = TC–(HDL–TG/5) and VLDL = TG/5. The atherogenic index (AI), atherogenic coefficient (AC), cardiac risk ratio (CRR), and coronary artery index (CAI) were calculated using the equations of Ikewuchi and Ikewuchi [19 (link)]. Additionally, serum levels of adiponectin, insulin, and leptin were measured using ELISA assays (Sigma-Aldrich, St. Louis, MO, USA). The homeostasis model assessment of insulin resistance (HOMA-IR) index was calculated as (fasting serum insulin × fasting serum glucose)/22.5 [20 (link)].