Journal: PLOS Pathogens

Article Title: Regulatory T cell-like response to SARS-CoV-2 in Jamaican fruit bats ( Artibeus jamaicensis ) transduced with human ACE2

doi: 10.1371/journal.ppat.1011728

Figure Lengend Snippet: Four bats were transduced with human ACE2 and infected 5 days later. After 12 days, splenocytes were recovered for AIM testing and gene expression analsysis. A. Splenocytes from hACE2-transduced bats 12 days post infection with SARS-CoV were cultured with or without SARS-CoV-2 nucleocapsid peptide library for 6 hours in the presence of anti-CD40 blocking antibody (AIM testing) or 24 hours without anti-CD40 blocking antibody (cellular RNA). B. Cells were stained with anti-CD154 and anti-CD4 and analyzed by flow cytometry. Violin plots for the individual bats, plus the pooled sample and rank data. In the presence of peptide, CD154 expression was significantly increased on splenocytes (red) compared to no peptide controls (blue) (Wilcoxon rank sum test). C. Heat map of splenocyte qPCR (ΔΔCq) gene expression in response to peptide stimulation determined 3 of the 4 bats expressed a profile consistent with a regulatory T cell response. D. Peptide-induced gene expression changes in splenocyte cultures of bats 1, 2 and 4 from panel C; IL-10, TGFβ and CD4 gene expression were significantly elevated, whereas CXCR4 was nearly significant (two-tailed t-test).

Article Snippet: Cells were then washed and stained with anti-Jamaican fruit bat CD4 (1:100 dilution) and anti-human CD154 (5C8, Tonbo/Cytek, 1:20 dilution).

Techniques: Transduction, Infection, Gene Expression, Cell Culture, Blocking Assay, Staining, Flow Cytometry, Expressing, Two Tailed Test