Costar 24 well tissue culture plates

1.75 mL of the heparinized blood was used to set up whole‐blood cultures for evaluation of in vitro production of cytokines (IL‐5, IL‐10, IL‐13, IFN‐γ) as previously described.16 Briefly, blood was diluted 1:5 in culture media (RPMI 1640, 1% L‐glutamine, 1% Penicillin‐streptomycin). 1.5 mL per well of this mixture was used for whole‐blood cultures in Corning^® Costar^® 24‐well tissue culture plates (Corning Inc., New York), with either no added stimulant or one of the following potential stimulants: phytohemagglutinin‐P (PHA; Sigma‐Aldrich, St. Louis, MO) at a final concentration of 2.5 μg/mL, schistosome soluble worm antigen preparation (SWAP) 10 μg/mL final concentration, soluble egg antigen (SEA) at final concentration of 5 μg/mL.17 The cultures were maintained for either 3 days (PHA and controls) or 5 days (antigens and controls) in a 5% CO₂ incubator at 37°C. Culture supernatant fluids were then harvested into cryovials and stored frozen (−20°C) until assayed together later for cytokine levels. Cytokine levels were measured using Duoset ELISA kits (R&D systems, Minneapolis, MN) as per manufacturer's protocol, with modifications as follows: (i) the detection antibody was left to incubate in the plate for 1 hour and (ii) an 8‐point standard curve was used instead of 7, with the lowest standard being 0 pg/mL.

THP-1 cells (3.5 × 105) were seeded on rounded cover slips in Costar^® 24-well tissue culture plates (Corning, Corning, NY, USA) and treated with phorbol 12-myristate 13-acetate (PMA, 100 nM; Sigma) in growth medium. After differentiation, the cells were incubated with oxLDL (200 μg/mL) for 24 h. Lipid uptake by macrophages was quantified by ORO staining.