Journal: Signal Transduction and Targeted Therapy

Article Title: Analogous comparison unravels heightened antiviral defense and boosted viral infection upon immunosuppression in bat organoids

doi: 10.1038/s41392-022-01247-w

Figure Lengend Snippet: Bat organoids elicited a more robust antiviral response to Poly(I:C) treatment. Bat and human intestinal organoids were sheared mechanically and treated with 10 μg/ml Poly(I:C) or mock-treated with DMSO. a , b Induction of IFNs (left) and ISGs (right) in bat intestinal organoids ( a ) and human intestinal organoids ( b ) at the indicated hours after Poly(I:C) treatment. Results show the log2-fold change of GAPDH-normalized expression level in the treated organoids relative to mock-treated organoids. Data represent the mean and s.d. of a representative experiment in organoids from a bat and human donor, n = 3. c Culture media from the treated or mock-treated human intestinal organoids at the indicated hours were applied to ELISA to measure concentrations of IFNL1 and IFNL3. Data represent the mean and s.d. of a representative experiment in one line of human organoids, n = 3. Two-tailed unpaired Student’s t test. d Poly(I:C) treated or mock-treated human intestinal organoids were collected at the indicated hours post-treatment and subjected to Western blot to detect human ISG15. e , f Bat and human intestinal organoids were sheared and incubated with 10 μg/ml Poly(I:C) Fluorescein in triplicate for 2 h and 6 h. The organoids were then dissociated and applied to flow cytometry to detect the percentage at 2 h post treatment ( e ) and mean fluorescence intensity (MFI, f ) of Fluorescein-positive cells at 2 and 6 h post treatment. g , h Induction of TNF-a and IL6 ( g ) and IP10 ( h ) in bat and human intestinal organoids at the indicated hours after treatment. Results show the fold change of GAPDH-normalized expression level in Poly(I:C)-treated organoids relative to mock-treated organoids. Data represent the mean and s.d. of a representative experiment in organoids from a bat and human donor, n = 3

Article Snippet: Cell-free media were harvested from treated and mock-treated human intestinal organoids for measuring the amount of human IFNL1 and IFNL3 using ELISA kits (R&D Systems, DY7246, D28B00).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Two Tailed Test, Western Blot, Incubation, Flow Cytometry, Fluorescence

Journal: Signal Transduction and Targeted Therapy

Article Title: Analogous comparison unravels heightened antiviral defense and boosted viral infection upon immunosuppression in bat organoids

doi: 10.1038/s41392-022-01247-w

Figure Lengend Snippet: TLR3 and RLR signaling pathways mediate antiviral responses in bat and human organoids. a A schematic graph outlines the experimental procedure for panels b – g . Bat and human intestinal organoids were pretreated with BX795 (0, 0.1 and 1 μM) or CYT387 (0, 0.1, and 1 μg/ml) overnight. After mechanical shearing, the organoids were incubated with or without 10 μg/ml Poly(I:C), together with the initial concentrations of BX795 or CYT387. At the indicated hours after Poly(I:C) treatment, the organoids were harvested and subjected to RT-qPCR assay to examine mRNA expression levels of bat and human IFNL1 and IFNL3; cell-free media from human organoids were applied to ELISA to detect IFNL1 and IFNL3. Data represent the mean and s.d. of a representative experiment in organoids from a bat and human donor, n = 3. Ordinary one-way ANOVA with Tukey’s multiple comparison test. b , c Induction of IFNL1 (left) and IFNL3 (right) in bat intestinal organoids treated with BX795 ( b ) and CYT387 ( c ). Results show the fold change of GAPDH-normalized expression level in the Poly(I:C)-treated organoids relative to mock-treated organoids. d , e Induction of IFNL1 and IFNL3 in human intestinal organoids treated with BX795 ( d ) and CYT387 ( e ). IFNL1 ( f ) and IFNL3 ( g ) secretion from the human organoids at 4 h post-stimulation. h A schematic graph describes the experimental procedure for panels i and j . Bat and human intestinal organoids were pretreated with 1 μg/ml CYT387 or DMSO overnight. After mechanical shearing, the organoids were incubated with 1 μg/ml CYT387 or DMSO with or without 10 μg/ml Poly(I:C) for 4 h. Cell-free media were then harvested and subjected to PRM-MS to analyze bat IFNL3 ( i ) and ISG15 of bat and human ( j ). The dotted lines represent the detection limit. Data represent the mean and s.d. of a representative experiment in organoids from a bat and human donor, n = 3. Two-tailed unpaired Student’s t test

Article Snippet: Cell-free media were harvested from treated and mock-treated human intestinal organoids for measuring the amount of human IFNL1 and IFNL3 using ELISA kits (R&D Systems, DY7246, D28B00).

Techniques: Protein-Protein interactions, Incubation, Quantitative RT-PCR, Expressing, Enzyme-linked Immunosorbent Assay, Comparison, Two Tailed Test

Journal: Frontiers in Immunology

Article Title: Human Placental Trophoblasts Infected by Listeria monocytogenes Undergo a Pro-Inflammatory Switch Associated With Poor Pregnancy Outcomes

doi: 10.3389/fimmu.2021.709466

Figure Lengend Snippet: Top 50 Induced Genes.

Article Snippet: IFNL1, IFNL2, and IFNL3 ELISAs were performed using the human IL-29 (DY7246), IL-28A (DY1587), and IL-28B (DY5259) DuoSet ELISA kits (R&D Systems) per the manufacturer’s instructions.

Techniques: