Figure 6.
Treatment with baricitinib and remdesivir increases the rate of viral clearance in lung-infiltrating monocytes
(A) Schematic of the experimental setup to quantify replication of the virus in monocytes. HLE-ALI cells were infected with SARS-CoV-2 at an MOI 0.1 for 48 h, after which 106 monocytes were transmigrated across the infected epithelium toward LTB4 (100 nM) and CCL2 (250 pg/mL) for 24 h, in the absence/presence of remdesivir (1 μM) and/or baricitinib (1 μM). Monocytes were washed and purified by negative depletion using anti-CD326 beads to remove contaminating epithelial cells and placed into medium with no drug, remdesivir (1 μM), and/or baricitinib (1 μM) for 0–72 h. All conditions contained 0.1% v/v DMSO.
(B) Quantification of total SARS-CoV-2 genome copies in the monocytes.
(C) Quantification of the amount of virus in the extracellular fluid at each time point.
(D) Quantification of the N-subgenome in monocytes at each time point. Data were normalized to 18S rRNA in the untreated condition at 0 h using the delta delta Ct method.
(E) The extracellular fluid was layered onto VeroE6 cells to perform a plaque assay from which a TCID50 was calculated. The positive control was the direct application of 2.5 × 104 genome copies of SARS-CoV-2 to the cells.
(F–H) Inflammatory mediators were measured using an electrochemiluminescent assay.
All statistics were calculated with a two-way ANOVA, main effects model in Prism with Geisser-Greenhouse correction applied. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001. Shown are median and interquartile range.