Journal: ACS Biomaterials Science & Engineering

Article Title: An Engineered Probiotic Produces a Type III Interferon IFNL1 and Reduces Inflammations in in vitro Inflammatory Bowel Disease Models

doi: 10.1021/acsbiomaterials.2c00202

Figure Lengend Snippet: Effect of EcN-gIFNL1 on regulatory T (Treg) cell populations. Flow cytometric analysis showed a synergistic effect of EcN-gIFNL1 on enhancing the population of CD4 + CD25 + Foxp3 + induced Treg (iTreg) cells. (A) Naïve CD4 + T cells were cultured for 24 h under inflammatory conditions and treated for 10 h with the indicated treatments. (B) Naïve CD4 + T cells were cocultured with primary IECs under inflammatory conditions and subsequently treated for 10 h with the indicated treatments. The concentration of IL-2 in the cell supernatants was analyzed through a multiplex assay. Similarly, flow cytometric analysis of the T cells in the coculture model revealed a synergistic effect of EcN-gIFNL1 on the enhancement of Treg cells. (C) Plots of iTreg cells (CD25 + Foxp3 + ) and effector Treg cells (CD25 hi Foxp3 hi ) were boxed and labeled accordingly. Bar graphs of iTreg cells (D) and effector Treg cells (E) were derived accordingly. n = 6 biological replicates; *, p < 0.05, **, p < 0.01 (Student’s t test).

Article Snippet: Cells in the basal compartments in all coculture setups were stained with surface markers FITC-labeled anti-CD4 (Cell Signaling Technology), APC-labeled anti-CD25 (Cell Signaling Technology), PE-Cy7-labeled anti-CD127 (eBioScience), and Treg-specific transcription factor PE-labeled anti-FoxP3 (eBioscience) antibodies for identification of Treg cells.

Techniques: Cell Culture, Concentration Assay, Multiplex Assay, Labeling, Derivative Assay