Amphotericin

A murine HPV 16-transformed BMK-16/myc cell line, which produces tumors in immunocompetent mice 13 (link),14 (link), was established by co-transformation of baby Balb/c kidney cells with the c-myc gene and the HPV 16 genome. The cell line was cultured in Dulbecco's Modified Eagles Medium (DMEM) supplemented with 10% of fetal bovine serum (FBS) (Gibco, ThermoFisher Scientific, Waitham, MA, USA), ampicillin, streptomycin and amphotericin (Caisson), at 37ºC in 95% humidity with 5% CO₂. IFN-τ ovine recombinant protein was obtained through Prospec, Rehovot Israel. Luna Universal Probe one-step RT-qPCR kit (New England Biolabs, Ipswich, MA, USA) primers were used to detect MHC Class I, MX, and IP10 was synthetized at the Biotechnology Institute in Mexico.

hBMVECs were grown in DMEM/F12 media (Invitrogen) containing 10 mm HEPES, 13 mm sodium bicarbonate (pH 7), 10 mmol/l L-glutamine and supplemented with 10% heat-inactivated fetal bovine serum (FBS; Thermo Fisher Scientific), penicillin and streptomycin (100 μg/ml each, Life Technologies), 1% amphotericin (Caisson Labs), endothelial cell growth supplement (ECGS; 50 mg/ml; BD Bioscience), heparin (100 mg/ml; Sigma-Aldrich Co, Ltd; Abdul Muneer et al., 2012 (link), 2018 (link)). hBMVECs were plated on rat tail collagen Type I (0.1 mg/ml) coated six-well Bioflex culture plates (Flexcell International Corp) at a density of 250,000 cells/well (Patel et al., 2017 (link); Abdul-Muneer et al., 2018 (link)). The cells were fed by changing the medium every 2 d and grown until tight monolayers were formed in ∼6–8 d.