Lectin Switching During Dengue Virus Infection

Dejnirattisai, Wanwisa; Webb, Andrew I.; Chan, Vera; Jumnainsong, Amonrat; Davidson, Andrew; Mongkolsapaya, Juthathip; Screaton, Gavin

The dengue-2 strain, 16681, was grown in C6/36 cells, Vero cells, and monocyte-derived dendritic cells (DCs). Cell-free supernatants were used either neat or after concentration by ultracentrifugation at 45,000 rpm for 4 h at 4°C, and the virus pellet was resuspended in 1.5% fetal bovine serum (FBS)/ Leibovitz L-15. To concentrate large volumes of low-titer DENV supernatant, DENV were precipitated with 10% Polyethylene glycol 8000 (Sigma) before ultracentrifugation. U937 were maintained in 10% FBS/Roswell Park Memorial Institute medium (RPMI). NIH/3T3, and DC-SIGN and L-SIGN NIH/3T3 cell lines were obtained through the AIDS Research and Reference Reagent Program (Division of AIDS, National Institute of Allergy and Infectious Diseases, from Drs Thomas D. Martin and Vineet N. Kewal Ramani) and maintained in 10% FBS/ Dulbecco's Modified Eagle's medium. All media were supplemented with 2 mmol/L L-glutamine, 100 U/mL penicillin, and 100 U/mL streptomycin.