Amphotericin b

Vero cells (ATCC, Manassas, VA, USA) were grown in EMEM (ATCC) supplemented with 10% fetal bovine serum (Gemini Bio-Products), penicillin/streptomycin (VWR), gentamicin (Sigma Aldrich), and amphotericin B (Quality Biological). ZIKV strain MR766 was added to Vero cells at MOI of 0.1 and incubated for 4–6 days. The supernatants were centrifuged at 1,500 rpm for 5 min and filtered (0.45 μm) before being concentrated via SnakeSkin dialysis tubing 3.5K MWCO (Thermo Scientific) in polyethylene glycol 8000 powder (Alfa Aesar) until all liquid was drawn out. The tubing was then placed in PBS overnight at 4°C. The reconstituted ZIKV was then aliquoted and stored at −80°C. ZIKV titers were determined using plaque assays on Vero cells as previously described (39 ). Briefly, ZIKV stocks were serially diluted and adsorbed to confluent monolayers of Vero cells. After 3 hours, the inoculum was removed, and cells were overlaid with semisolid medium containing 1% carboxymethyl cellulose (Sigma Aldrich). Cells were further incubated for 5 days, fixed with 4% paraformaldehyde (Electron Microscopy Sciences), and stained with 0.5% aqueous crystal violet solution (Sigma Aldrich) for plaque visualization. Titers were expressed as plaque forming units (PFU) per milliliter.

The panel of hematological cancer cell lines used in this study consisted of BJAB (Burkitt’s Lymphoma), CCRF-CEM (Acute Lymphoblastic Leukemia, ATCC: 62848170), HL-60 (Acute Myelocytic Leukemia, ATCC: 62690063), Jurkat (Acute Lymphocytic Leukemia, ATCC: 62729601), Nalm-6 (Acute Lymphoblastic Leukemia) and Ramos (Burkitt’s Lymphoma, ATCC: 62927031). Five of the cell lines were cultured in RPMI-1640 medium (Corning 10–040-CV) supplemented with 10% heat-inactivated fetal bovine serum (FBS; Atlanta Biologicals Cat No. S11150) and a mixture of 25 μg/ml amphotericin B, 1000 U/ml penicillin, and 1000 μg/ ml streptomycin (Quality Biological 120–096–711). The HL-60 cells were supplemented with 20% FBS in RPMI-1640 medium and the antibiotics mentioned above. The non-cancerous cell lines used were Hs27 (human foreskin fibro-blast, ATCC: 3681588) cultured in DMEM medium supplemented with 10% heat-inactivated FBS and a mixture of 25 μg/ml amphotericin B, 1000 U/ml penicillin and 1000 μg/ml streptomycin, and MCF-10A (human breast epithelium, ATCC: CRL-10317) cultured in DMEM/F12 medium supplemented with FBS and antibiotics as mentioned above, as well as 20 ng/ml epidermal growth factor, 0.5 μg/ ml hydrocortisone (Sigma-Aldrich, H0888) and 10 μg/ ml insulin (Sigma-Aldrich, I1882). All cells were incubated at 37 °C in a humidified atmosphere containing 5% CO₂.