Simulation and analysis of <i>Helicobacter pylori</i> colonization.

<p>Time course simulation with calibrated <i>H</i>. <i>pylori</i>-host interaction model displaying <i>H</i>. <i>pylori</i> burden (A), macrophage and cytokine responses (B) and T cell responses (C). Local sensitivity analysis of model displaying positive and negative effects of parameters on model species H. pylori (D), IL10 (E), damaged epithelial cells (F). Boxed region contains parameters associated with Mreg model species. Specific model species of interest are indicated by symbols: & marks column associated with HP model species, * marks column associated with IL10 model species and # marks column associated with Edamaged model species. Global sensitivity analysis displaying parameter sensitivity ranges on the differentiation of regulatory macrophages (G). Parameters associated with cytokines are indicated by (*) and parameters associated with <i>H</i>. <i>pylori</i> are indicated by (#).</p

Network of intracellular signaling pathways controlling regulatory macrophage differentiation.

<p>Systems Biologt Markup Language (SBML)-compliant network diagram displaying the three main pathways, CSF1R, CX3CR1, and IL-10R, controlling cytokine production and macrophage phenotype commitment. While interconnected, the three signaling receptors have dominant effects on arms within the network. The CSF1R/M-CSF pathway integrates DAP12 and NFAT activity and is closely associated with the inflammatory Traf/NF-kB pathway as well. The IL-10R pathway is modified by calcium signaling and the transcriptional activities of FOXP1 and NCOR2. The CX3CR1 pathway combines cAMP/PKA pathways with modification to Akt and inflammatory signaling.</p

Simulation, analysis and validation of intracellular macrophage network model.

<p>Global sensitivity analysis displaying parameter sensitivity ranges on the differentiation of regulatory macrophages (A). Parameters are grouped by associated model species: NFkB-associated (purple), LANCL2-associated (red), PKA-associated (blue), IL10-associated (green). Predicted impairment of regulatory macrophage differentiation following simulated loss of LANCL2 (B). Fold changes in molecule production or activation following simulated loss of LANCL2 compared to wild-type simulation (C). mRNA expression of LANCL2 in whole stomach (D) and cultured bone marrow derived macrophages (E), normalized to expression of beta-actin. mRNA expression of Ncor2 (F) and Foxp1 (G), Klf4 (H), and Il10 (I) within the stomach of <i>H</i>. <i>pylori</i> SS1-infected and uninfected LANCL2fl/fl; LysCre- and LANCL2fl/fl LysCre+ mice at three weeks post-infection. NFkB p65 activity in whole stomach tissue homogenate of <i>H</i>. <i>pylori</i> SS1-infected and uninfected LANCL2fl/fl; LysCre- and LANCL2fl/fl LysCre+ mice at three weeks post-infection (J). <i>p</i>-values less than 0.05 are considered significant and marked by an asterisk (*), (n = 7).</p

Network model of <i>Helicobacter pylori</i>-host interactions.

<p>Systems biology markup language (SBML)-compliant network diagram of four compartment <i>H</i>. <i>pylori</i> response model. Cell types include <i>H</i>. <i>pylori</i> (HP), tolerogenic bacteria (TolB), epithelial cells (E), damaged epithelial cells (Edamaged), immature dendritic cells (iDC), effector dendritic cells (eDC), tolerogenic dendritic cells (tDC), monocytes (Monocytes), regulatory macrophages (M_reg), naïve T cells (nT), T helper 1 (Th1), T helper 17 (Th17), induced T regulatory (iTreg), type 1 regulatory (Tr1), IFNγ (IFNg), and IL10 (IL10).</p

Oral treatment with NSC61610 on the distribution of macrophages in colonic lamina propria.

<p>Colonic lamina propria lymphocytes (LPL) were immunophenotyped to identify F4/80+CD11b+ macrophage subsets through flow cytometry. Data are represented as mean ± standard error (n = 10). Bars with an asterisk indicate that a treatment is significantly different from its control (<i>P<0.05</i>).</p

Effect of LANCL2 disruption and cAMP inhibition on PPAR γ activation in RAW 264.7 macrophages.

<p>Cells were cotransfected with a pTK.PPRE3x luciferase reporter plasmid driven by the PPRE-containing Acyl-CoA oxidase promoter with or without LANCL2 siRNA. Then, cells were treated with vehicle (DMSO) or NSC61610 (2.5 µM), the adenylate cyclase-specific inhibitor 2′5′-dideoxyadenosine (10 µM). Luciferase activity was normalized to pRL activity in the cell extracts and relative luciferase activity was calculated a ratio of the activity in the treatment wells to control wells. Data are represented as mean ± standard error. Points with an asterisk indicate that a treatment is significantly different from its control (<i>P<0.05</i>).</p

Oral treatment with NSC61610 ameliorates inflammatory lesions in mice with inflammatory bowel disease.

<p>Mice were challenged with 2.5% dextran sodium sulfate in the drinking water for 7 days. Representative photomicrographs from the control (A–B) and NSC61610 treatment (C–E) groups are illustrated. Colonic specimens underwent blinded histological examination and were scored based on epithelial erosion (F), mucosal wall thickening (G), and leukocyte infiltration (H). Data are represented as mean ± standard error (n = 10). Bars with an asterisk indicate that a treatment is significantly different from its control (<i>P<0.05</i>).</p

Lanthionine synthetase C-like 2 (LANCL2) and NSC61610 <i>in silico</i>.

<p>(A) The homology model of human LANCL2 is shown in Cartoon representation with coloring according to secondary structure. Purple: alpha helix; Blue: other helix; Yellow: bridge_beta; Cyan: turn; Green: coil. (B) 2-D structure of NSC61610. (C) Representative binding mode of the most stable docked orientation of NSC61610 with LANCL2. The LANCL2 model is shown in ribbon mode. NSC61610 pose generated by AutoDock Vina is colored in cyan and the one generated by AutoDock is colored in orange. Selected residues of LANCL2 (blue) are depicted by stick-and-ball models and colored by atom types. Amino acid residues surrounding NSC61610 are labeled. (D) Representative binding modes of the docked orientation of abscisic acid (ABA) and NSC61610 with LANCL2. LANCL2 is shown in a ribbon mode. NSC61610 (orange) and ABA (magenta) are shown in stick-and-ball model. Selected residues of LANCL2 surrounding both NSC61610 and ABA are depicted by stick-and-ball model and labeled. The images were rendered in Visual Molecular Dynamics (VMD).</p

Modulation of colonic gene expression by oral treatment with NSC61610.

<p>Mice were challenged with 2.5% dextran sodium sulfate in the drinking water for 7 days. Colonic mRNA expression of peroxisome proliferator-activated receptor γ (PPAR γ) (A), monocyte chemoattractant protein-1 (MCP-1) (B), and interleukin-6 (IL-6) (C) were assessed by quantitative real-time RT-PCR. Data are represented as mean ± standard error (n = 10). Bars with an asterisk indicate that a treatment is significantly different from its control (<i>P<0.05</i>).</p

Oral treatment with NSC61610 ameliorates experimental inflammatory bowel disease.

<p>Mice were challenged with 2.5% dextran sodium sulfate in the drinking water for 7 days. Disease activity index (DAI), a composite score reflecting clinical signs of the disease (i.e. perianal soiling, rectal bleeding, diarrhea and piloerection), was assessed daily. Panel A illustrates the effect of NSC61610 on disease severity in mice with colitis. Panels B–D illustrate the effect of NSC61610 on macroscopic inflammatory lesions in the colon (B), spleen (C), and mesenteric lymph nodes (MLN) (D). Data are represented as mean ± standard error (n = 10). In figure A, data points with asterisks are significantly different from control and data points with two asterisks are significantly different from those with one asterisk (<i>P<0.05</i>). In figure B–D, bars with an asterisk indicate that a treatment is significantly different from its control (<i>P<0.05</i>).</p