Glucose but not galactose culturing of IFN-γ primed cells reduces VACV and HSV-1 replication.

(A) Western blot analysis of VACV proteins–early (SSB) and late (A27)–in A549 cells primed with either IFN-α or IFN-γ, grown in glucose or galactose. (B) qPCR of VACV transcripts–early (I3L) and late (F17R) genes (N = 3). (C) Top: Quantification of plaque assay for VACV-Luc-GFP infectious units from A549 cells (N = 3). Bottom: representative image of plaque assay with various dilutions of viral titers (10⁻¹–10⁻³). (D) Western blot analysis for HSV-1 viral immediate early (ICP27) and tegument proteins (VP16) in A549 cells. (E) Quantification of plaque forming units from HSV-1-GFP A549 infected cells primed with either glucose or galactose as well as IFN-α or IFN-γ (N = 3). (F) VSV (luciferase) replication assay from IFN-pretreated A549 cells grown in glucose or galactose. Bottom: images showing presence or absence of cytopathic effect (with interferon priming). Plaque forming unit: P.F.U. Glc: glucose (25 mM). Gal: galactose (10 mM). Loading control for western blot and qPCR: β-actin. For viral infection, MOI = 0.01 for VACV-Luc-GFP and VSV-Luc; MOI = 1 for HSV-1-GFP. Concentration for IFN-α and IFN-γ: 1000 units/mL. Statistical analysis was performed using an unpaired t-test in GraphPad Prism 9.5.1: n.s. not significant, * P ≤ 0.05, ** P < 0.01, *** P ≤ 0.001, **** P ≤ 0.0001.