昭和20年代, 教室の日常と研究余話(第922回千葉医学会例会・第13回千葉精神科集談会)

<p>Unique MAA Arcs and Mechanistic Genes in Tight Junction Pathway.</p

Pathway Score Heat Map for Cell Communication Related Pathways.

<p>This heat map shows the repressed state of activity for the Tight Junction (TJ) and Trefoil Factors (TF) pathway in comparison to the Gap Junction, Integrin-mediated Cell Adhesion and Cell Adhesion Molecules pathways. Note that the TJ and TF pathways have a complex activation pattern. The TJ is tri-phasic in that it is highly repressed in the early stage, becomes moderately activated at 60 and 120 min p.i, and then becomes repressed at 240 min p.i. The TF pathway is bi-phasic in that it is highly repressed in the early stage but becomes moderately activated at 120 and 240 min p.i. The darker red gradients indicate higher Bayesian activation scores (more up-regulated gene expression within the pathway) while the darker green gradients indicate more repressed pathway activity (more down-regulated gene expression).</p

Systems Analysis of Early Host Gene Expression Provides Clues for Transient <i>Mycobacterium avium</i> ssp <i>avium</i> vs. Persistent <i>Mycobacterium avium</i> ssp <i>paratuberculosis</i> Intestinal Infections

<div><p>It has long been a quest in ruminants to understand how two very similar mycobacterial species, <i>Mycobacterium avium</i> ssp. p<i>aratuberculosis</i> (MAP) and <i>Mycobacterium avium</i> ssp. <i>avium</i> (MAA) lead to either a chronic persistent infection or a rapid-transient infection, respectively. Here, we hypothesized that when the host immune response is activated by MAP or MAA, the outcome of the infection depends on the early activation of signaling molecules and host temporal gene expression. To test our hypothesis, ligated jejuno-ileal loops including Peyer’s patches in neonatal calves were inoculated with PBS, MAP, or MAA. A temporal analysis of the host transcriptome profile was conducted at several times post-infection (0.5, 1, 2, 4, 8 and 12 hours). When comparing the transcriptional responses of calves infected with the MAA versus MAP, discordant patterns of mucosal expression were clearly evident, and the numbers of unique transcripts altered were moderately less for MAA-infected tissue than were mucosal tissues infected with the MAP. To interpret these complex data, changes in the gene expression were further analyzed by dynamic Bayesian analysis. Bayesian network modeling identified mechanistic genes, gene-to-gene relationships, pathways and Gene Ontologies (GO) biological processes that are involved in specific cell activation during infection. MAP and MAA had significant different pathway perturbation at 0.5 and 12 hours post inoculation. Inverse processes were observed between MAP and MAA response for epithelial cell proliferation, negative regulation of chemotaxis, cell-cell adhesion mediated by integrin and regulation of cytokine-mediated signaling. MAP inoculated tissue had significantly lower expression of phagocytosis receptors such as mannose receptor and complement receptors. This study reveals that perturbation of genes and cellular pathways during MAP infection resulted in host evasion by mucosal membrane barrier weakening to access entry in the ileum, inhibition of Ca signaling associated with decreased phagosome-lysosome fusion as well as phagocytosis inhibition, bias toward Th2 cell immune response accompanied by cell recruitment, cell proliferation and cell differentiation; leading to persistent infection. Contrarily, MAA infection was related to cellular responses associated with activation of molecular pathways that release chemicals and cytokines involved with containment of infection and a strong bias toward Th1 immune response, resulting in a transient infection.</p></div

Interleukin Gene Score Heat Map by Time Point Post Inoculation.

<p>The heat map shows a number of both up-regulated and down-regulated genes (15–240 minutes post-inoculation). The heat map shows that much of the up-regulation is short-lived for many of these genes and reversed expression direction or minimally expressed in later time points. Red indicates an activated state while green indicates repression and grey is no expression change from control.</p

Toll-like Receptor Related Gene Score Heat Map by Time Point Post-Inoculation.

<p>The heat map shows a number of up-regulated genes occurring in the early stage (15–60 minutes post-inoculation) as indicated by the darker red boxes as well as down-regulated genes as indicated by the darker green boxes.</p

Primers for Real Time – PCR Analysis of Genes in <i>B. melitensis</i>-Infected Bovine Peyer's Patch.

<p>Primers for Real Time – PCR Analysis of Genes in <i>B. melitensis</i>-Infected Bovine Peyer's Patch.</p

Genes Significantly Expressed in the TLRS Pathway (<i>z</i>-score |2.24|).

<p>The table lists significantly perturbed (activated or repressed) genes at any time among the 5 different time points (0.25 h through 4 h p.i.).</p

Graphic Representation of Bovine Genes Differentially Expressed throughout the Experiment.

<p>(DE  =  differentially expressed). Solid bars represent genes up-regulated; open bars represent genes down-regulated.</p

Significantly Altered Genes (Bayesian <i>z</i>-score |2.24|) in the Extra-Cellular Membrane (ECM) pathway.

<p>The table lists significantly perturbed (activated or repressed) genes at any time among the 5 different time points (0.25 h through 4 h p.i.).</p

Heatmap of Significantly Perturbed Pathways Scores in Bovine Peyer's Patch Infected with <i>B. melitensis</i>.

<p>Thirty-seven pathways were identified as significantly perturbed (97.5% confidence) from the early stage of infection (15–60 m.p.i.). The darker red gradients indicate higher Bayesian activation scores (more up-regulated gene expression within the pathway) while the darker green gradients indicate more repressed pathway activity (more down-regulated gene expression).</p