Journal: Frontiers in Immunology

Article Title: Faecalibacterium prausnitzii Skews Human DC to Prime IL10-Producing T Cells Through TLR2/6/JNK Signaling and IL-10, IL-27, CD39, and IDO-1 Induction

doi: 10.3389/fimmu.2019.00143

Figure Lengend Snippet: DC exposure to F. prausnitzii promotes the production of immunoregulatory molecules. (A) : IL-10 (Ctrl and F n = 26, M88 n = 3, M89 n = 9), (B): IL-27 (Ctrl, F and M89 n = 9, M88 n = 6) and (C): IL-6 (Ctrl and F n = 12, M89 n = 6) -secretion by DC exposed or not to indicated bacteria during the last 24 h. (D–F) : expression of PDL-1 and CD39 ( n = 15–29) or IDO-1 ( n = 18–25) by DC exposed or not to indicated bacteria for the last 24 or 48 h. (G–I) : IL-10 ( n = 6), IL-27 ( n = 9), and IL-6 ( n = 3) secretion by myeloid DC maintained for 24 h in culture with or without F. prausnitzii . Results as mean ± sem. Wilcoxon test. * p < 0.05, ** p < 0.005, *** p < 0.0005, **** p < 0.00005.

Article Snippet: For blocking experiments, we used rat neutralizing Ab to IL-10 (10 μg/ml, BD), IL-10R (10 μg/ml, R&D Systems), TLR2, TLR6, and Rat IgG control (20 μg/ml, InvivoGen), the JNK inhibitor SP600125 (10 μM, Abcam), the inhibitor of the CD39 ecto-nucleotidase activity Pom1 (10 μM, Tocris) and the IDO1 inhibitor (400 μM, Sigma Aldrich).

Techniques: Bacteria, Expressing

Journal: Frontiers in Immunology

Article Title: Faecalibacterium prausnitzii Skews Human DC to Prime IL10-Producing T Cells Through TLR2/6/JNK Signaling and IL-10, IL-27, CD39, and IDO-1 Induction

doi: 10.3389/fimmu.2019.00143

Figure Lengend Snippet: TLR2/6- and JNK-dependent modulation of DC function by F. prausnitzii . (A,B) : IL-10-secretion by d5-DC exposed or not to F. prausnitzii for the last 24 h, in the presence or not of neutralizing anti-TLR2 [ (A) n = 3] or anti-TLR6 Ab [ (B) n = 8]. (C–F) : LPS-induced IL-12p40 or p35 expression by DC exposed or not to F. prausnitzii at DC:bacterium ratios 1:1 (F or F1), 2:1 (F2-) or 5:1 (F5-), during the last 48 h in the presence or not of neutralizing anti-TLR2 [ (C) n = 6, (D) n = 5] or -TLR6 Ab [ (E) n = 3, (F) n = 3]. Secretion of IL-10 [ (G) n = 9), IL-27 [ (H) n = 14] or expression of PDL-1 [ (I) n = 9] or CD39 [ (J) n = 12] by DC exposed or not (Ctrl) to F. prausnitzii for the last 24 h, in the presence or not of the JNK inhibitor or its vehicle control DMSO. Paired t -test for n = 3, Wilcoxon test otherwise. * p < 0.05, ** p < 0.005, *** p < 0.0005.

Article Snippet: For blocking experiments, we used rat neutralizing Ab to IL-10 (10 μg/ml, BD), IL-10R (10 μg/ml, R&D Systems), TLR2, TLR6, and Rat IgG control (20 μg/ml, InvivoGen), the JNK inhibitor SP600125 (10 μM, Abcam), the inhibitor of the CD39 ecto-nucleotidase activity Pom1 (10 μM, Tocris) and the IDO1 inhibitor (400 μM, Sigma Aldrich).

Techniques: Expressing, Control

Journal: Frontiers in Immunology

Article Title: Faecalibacterium prausnitzii Skews Human DC to Prime IL10-Producing T Cells Through TLR2/6/JNK Signaling and IL-10, IL-27, CD39, and IDO-1 Induction

doi: 10.3389/fimmu.2019.00143

Figure Lengend Snippet: IL-27-dependent modulation of DC functions by F. prausnitzii . Expression of CD39 [ (A) n = 8], PDL-1 [(B) n = 9] or IDO1 [ (C) n = 3], and IL-10 secretion [ (D) n = 6] by DC incubated or not for the last 24 h with F. prausnitzii or with rhIL-27. IL-12p40 [ (E) n = 6] or p35 [ (F) n = 6] expression by DC incubated or not for the last 24 h with F. prausnitzii or with rhIL-27 and then stimulated overnight by LPS. Wilcoxon test. * p < 0.05, ** p < 0.005.

Article Snippet: For blocking experiments, we used rat neutralizing Ab to IL-10 (10 μg/ml, BD), IL-10R (10 μg/ml, R&D Systems), TLR2, TLR6, and Rat IgG control (20 μg/ml, InvivoGen), the JNK inhibitor SP600125 (10 μM, Abcam), the inhibitor of the CD39 ecto-nucleotidase activity Pom1 (10 μM, Tocris) and the IDO1 inhibitor (400 μM, Sigma Aldrich).

Techniques: Expressing, Incubation

Journal: Frontiers in Immunology

Article Title: Faecalibacterium prausnitzii Skews Human DC to Prime IL10-Producing T Cells Through TLR2/6/JNK Signaling and IL-10, IL-27, CD39, and IDO-1 Induction

doi: 10.3389/fimmu.2019.00143

Figure Lengend Snippet: CD39 and IDO1-dependent modulation of DC functions by F. prausnitzii . Secretion of IL-10 [ (A) n = 8] or IL-27 [ (B) n = 10] or expression of IDO1 [ (C) n = 4] or PDL-1 [ (D) n = 6] by DC incubated or not for the last 24 h with F. prausnitzii , following or not a 45 min-incubation with the CD39 inhibitor Pom1 (10 μM). (E) ( n = 3) LPS-induced IL-12-secretion by DC incubated, or not (Ctrl) with F. prausnitzii for 48 h, following or not a 45 min-incubation with the Pom1 inhibitor. (F,G) : ( n = 5) CD39 and PDL-1 expression by DC incubated (F), or not (Ctrl) with F. prausnitzii for 24 h, following or not a 45 min-incubation with the IDO1 inhibitor 1-MT. (A,B): Wilcoxon test, (E): Mann Whitney, (C,D,F,G) Paired t -test. * p < 0.05, ** p < 0.005, **** p < 0.00005.

Article Snippet: For blocking experiments, we used rat neutralizing Ab to IL-10 (10 μg/ml, BD), IL-10R (10 μg/ml, R&D Systems), TLR2, TLR6, and Rat IgG control (20 μg/ml, InvivoGen), the JNK inhibitor SP600125 (10 μM, Abcam), the inhibitor of the CD39 ecto-nucleotidase activity Pom1 (10 μM, Tocris) and the IDO1 inhibitor (400 μM, Sigma Aldrich).

Techniques: Expressing, Incubation, MANN-WHITNEY

Journal: Frontiers in Immunology

Article Title: Faecalibacterium prausnitzii Skews Human DC to Prime IL10-Producing T Cells Through TLR2/6/JNK Signaling and IL-10, IL-27, CD39, and IDO-1 Induction

doi: 10.3389/fimmu.2019.00143

Figure Lengend Snippet: Schematic representation of the interactions involved in the alteration of DC function following exposure to F. prausnitzii . IL-27 secretion induced by F. prausnitzii in a JNK dependent manner up-regulates CD39 expression. The ENTPDase activity of CD39 then contributes to IL-10 and IL-27 induction and up-regulates IDO1, in a feed-forward loop. IL-10 secretion also induced via the TLR2/6/JNK pathway limits the LPS-induced expression of CD83, which is affected by TLR2/6 triggering but not JNK blocking. Finally, PDL-1 expression relies on IDO1 enzymatic activity and, partly, on JNK and IL-27 signaling.

Article Snippet: For blocking experiments, we used rat neutralizing Ab to IL-10 (10 μg/ml, BD), IL-10R (10 μg/ml, R&D Systems), TLR2, TLR6, and Rat IgG control (20 μg/ml, InvivoGen), the JNK inhibitor SP600125 (10 μM, Abcam), the inhibitor of the CD39 ecto-nucleotidase activity Pom1 (10 μM, Tocris) and the IDO1 inhibitor (400 μM, Sigma Aldrich).

Techniques: Expressing, Activity Assay, Blocking Assay