IL-4/IL-13 polarization of macrophages enhances Ebola virus glycoprotein-dependent infection
Pmacs from Ifnar/Timd-4-/- mice were used in all studies. A) IL-4/IL-13 treated or untreated cells were infected with rVSV/EBOV GP (MOI = 0.1) or rVSV/G (MOI = 0.03) and infection was quantified by detection of GFP expression at 24 hours post infection. Expression is shown relative to unstimulated controls. B) IL-4/IL-13 treated or untreated cells were placed on ice and infected with rVSV/EBOV GP (MOI = 0.1). After 1 hour, cells were washed with PBS to remove unbound virus and RNA was isolated for qRT-PCR. Results are expressed as VSV-M RNA relative to HPRT control. C) IL-4/IL-13 treated or untreated cells were incubated with VLPs expressing EBOV GP and EBOV VP40 fused to β-lactamase. Entry was subsequently quantified by loading cells with CCF2-AM, a β-lactam-containing fluorescent substrate, and analyzed by flow cytometry to determine the relative amount of cleaved and un-cleaved species. D) IL-4/IL-13 treated or untreated cells were preincubated with 200ug/mL of mannan or GalNac for 1 hour. Cells were infected with rVSV/EBOV GP (MOI = 0.1) and GFP+ cells were quantified 24 hours post infection. Data are shown as mean ± S.D. Each experiment was performed at least 3 times. Statistical analysis was performed using Student’s t-test, *p<0.05.