5 research outputs found
TLR2/TLR4 regulate <i>F</i>. <i>nucleatum</i>-induced inflammatory cytokines through Tregs in vivo.
<p>(A, B and C) Production of IL-8, IL-1β and TNF-α in C57BL/6 wild-type mice and in hybrid, TLR2<sup>-/-</sup> and TLR4<sup>-/-</sup> mice infected with <i>F</i>. <i>nucleatum</i> for 1 week, as assessed by enzyme-linked immunosorbent assay (ELISA). (D) The number of <i>F</i>. <i>nucleatum</i> 16s rRNA gene copies in the gastric mucosa of C57BL/6 wild-type mice and in hybrid, TLR2<sup>-/-</sup> and TLR4<sup>-/-</sup> mice determined by qRT-PCR. (E) After transfer of Tregs from wild-type mice to TLR2<sup>-/-</sup> mice, the production of IL-8, IL-1β and TNF-α in TLR2<sup>-/-</sup> mice infected with <i>F</i>. <i>nucleatum</i> for 1 week and in the uninfected group was assessed by ELISA.</p
TLR2/TLR4 are involved in the <i>F</i>. <i>nucleatum</i>-induced inflammatory signaling pathway.
<p>(A) Signaling pathways of differentially expressed genes in Caco-2 cells infected with <i>F</i>. <i>nucleatum</i>. Pathway analysis was predominantly based on the KEGG database. An AP-value of <0.05 and an FDR of <0.05 in the two-sided Fisher’s exact test were considered statistically significant. The vertical axis represents the pathway category, and the horizontal axis represents the log 10 (<i>P</i> value) of these significant pathways. (B and C) Caco-2 cells or C57BL/6 mice were infected with <i>F</i>. <i>nucleatum</i> for 24 h or 1 week. The mRNA levels of TLR1, TLR2, TLR4, TLR5 and TLR6 were determined by qRT-PCR. (D) The correlation between TLR2/TLR4 and <i>F</i>. <i>nucleatum</i> 16s rRNA gene copies in human clinical specimens was examined by Spearman correlation analysis and found to be positive (TLR4/<i>F</i>. <i>nucleatum</i>, R = 0.324, <i>P</i> = 0.12; TLR2/<i>F</i>. <i>nucleatum</i>, R = 0.618, <i>P</i> = 0.006).</p
TLR2/TLR4 signaling modulates <i>F</i>. <i>nucleatum</i>-induced inflammation in vivo.
<p>(A) Histological inflammation scores (H&E staining) of the gastric mucosa of C57BL/6 wild-type mice and TLR2<sup>-/-</sup> and TLR4<sup>-/-</sup> mice with <i>F</i>. <i>nucleatum</i> infection for 1 week. The intensity of staining is shown in the right graph, and the data are expressed as the mean±SEM. (B) The body weight of C57BL/6 wild-type mice and TLR2<sup>-/-</sup> and TLR4<sup>-/-</sup> mice after <i>F</i>. <i>nucleatum</i> infection. (C, D and E) The production of IL-8, IL-1β and TNF-α in C57BL/6 wild mice and TLR2<sup>-/-</sup> and TLR4<sup>-/-</sup> mice infected with <i>F</i>. <i>nucleatum</i> for 1 week, as assessed by enzyme-linked immunosorbent assay (ELISA). (F) The number of <i>F</i>. <i>nucleatum</i> 16s rRNA gene copies in the gastric mucosa of C57BL/6 wild mice and TLR2<sup>-/-</sup> and TLR4<sup>-/-</sup> mice based on qRT-PCR.</p
TLR2/TLR4 activation induces Tregs and suppresses intestinal inflammation caused by <i>Fusobacterium nucleatum in vivo</i>
<div><p>Toll-like receptors (TLRs) 2 and 4 play critical roles in intestinal inflammation caused by <i>Fusobacterium nucleatum</i> (<i>F</i>. <i>nucleatum</i>) infection, but the role of TLR2/TLR4 in regulation of proinflammatory cytokines remains unknown. In this study, through microarray analysis and qRT-PCR, we showed that TLR2/TLR4 are involved in the <i>F</i>. <i>nucleatum</i>-induced inflammatory signaling pathway in Caco-2 cells, C57BL/6 mice and human clinical specimens. In TLR2<sup>-/-</sup> and TLR4<sup>-/-</sup> mice, <i>F</i>. <i>nucleatum</i> infection resulted in increased colonization of the bacteria and production of the proinflammatory cytokines IL-8, IL-1β and TNF-α. In addition, the ratio of Foxp3<sup>+</sup> CD4<sup>+</sup> T cells in the total CD4<sup>+</sup> T cells in TLR2<sup>-/-</sup> and TLR4<sup>-/-</sup> mice was less than that in wild-type mice, and the ratio in hybrid mice was more than that in knockout mice, which suggested that TLR2/TLR4 mediated the number of Tregs. Furthermore, it was observed that inflammatory cytokine levels were reduced in TLR2<sup>-/-</sup> mice after Treg transfer. Thus, these data indicate that TLR2/TLR4 regulate <i>F</i>. <i>nucleatum</i>-induced inflammatory cytokines through Tregs in vivo.</p></div
<i>F</i>. <i>nucleatum</i> infection increased the ratio of regulatory T cells in vivo.
<p>(A) Sixteen C57BL/6 wild mice were divided into two groups (n = 8). After eight C57BL/6 wild-type mice were infected with <i>F</i>. <i>nucleatum</i> for 1 week, FoxP3 and CD4 expression was analyzed with FACS after fixation and permeabilization, and a gating control was used. The <i>P</i> value (Mann–Whitney) is indicated. (B) Detection of Foxp3<sup>+</sup> CD4<sup>+</sup> T cells in total CD4<sup>+</sup> T cells in wild-type, TLR2<sup>-/-</sup> and TLR4<sup>-/-</sup> mice. (C) Detection of Foxp3<sup>+</sup> CD4<sup>+</sup> T cells in total CD4<sup>+</sup> T cells in wild-type, hybrid, TLR2<sup>-/-</sup> and TLR4<sup>-/-</sup> mice.</p