DSS-induced chronic colitis in CD69 KO mice.

<p>CD69 KO mice and wild-type (WT) mice were administered 2% DSS on days 0–5, 10–15, and 20–25. The body weight (A) and disease activity index (B) were monitored every day, and the values for body weight are expressed as the percentage of body weight on day 0. The data are presented as the means with SD (n = 5 per group). *p<0.01. Three independent experiments were performed with similar results.</p

Effect of <i>in vivo</i> treatment with an anti-CD69 monoclonal antibody (mAb) on DSS-induced colitis.

<p>(A) Wild-type (WT) BALB/c mice were treated with an anti-CD69 mAb or control hamster IgG on day 0. The survival of each group during DSS-induced colitis was recorded daily (n = 5 per group). (B, C) Changes in the disease activity index (B) and body weight (%) (C) over the course of DSS treatment in WT mice treated with the anti-CD69 mAb or control hamster IgG. The data are presented as the means (SD) (n = 5 per group). *p<0.01. **p<0.05. (D) Histological sections. Colons were taken on day 8 from control and DSS-exposed WT mice treated with the anti-CD69 mAb or control hamster IgG. Sections were fixed and stained with hematoxylin and eosin (original magnification 40×). Three independent experiments were performed with similar results.</p

The expression of cytokines, chemokines and their receptors in the colons of DSS-treated mice.

<p>(A) The expression of cytokines, chemokines and their receptor mRNA in the colons of DSS-treated mice. Mice were sacrificed on day 8 and the colon tissues were harvested. Whole colonic RNA was isolated, reverse-transcribed into cDNA, and the expression levels of IL-1β, IL-6, IL-10, CCR2, CCR3, CCL2, CCL4 and CCL5 were determined by real-time quantitative PCR. Data are expressed as the ratios of the target mRNA levels to the HPRT mRNA level (n = 5 per group). *p<0.05. Data are presented as the means (SD). The data are representative of three independent experiments. (B) The intracellular staining profiles of IL-10 in the lamina propria CD4 T cells (CD4⁺), CD8 T cells (CD8⁺), macrophages (CD11b⁺, Gr-1^int) and dendritic cells (CD11c⁺) are shown as the percentages of cells in each area. The ratios of IL-10⁺ cells/total cells of each cell subset are shown over the area bar. The ratios of IL-10⁺ cells in each subset/total cells of all subsets are shown under the area bar. The results are representative of three independent experiments.</p

DSS-induced colocecal damage was reduced in CD69 KO mice.

<p>(A) The colon length. On day 8, the colon length of DSS-exposed wild-type (WT) and CD69 KO mice and control mice was measured. The data are presented as the means (SEM) (n = 12 per group). *p<0.01. (B) A representative photograph showing the gross appearance of the colon from each group is shown. (C) Histological sections of inflamed colons. Colons were taken on day 8 from WT and CD69 KO mice that received DSS in drinking water. Sections were prepared and stained with hematoxylin and eosin (original magnification 40×). (D) Immunohistochemical staining of CD69-positive cells in the colonic tissues of DSS-treated WT mice (original magnification 300× and 600×). (E, H) Immunohistochemical staining of CD3-positive cells (E) and FoxP3-positive cells (H) in the colonic tissues of DSS-treated WT and CD69 KO mice (original magnification 200×). (F, I) Summary of the accumulation of CD3-positive cells (F) and FoxP3-positive cells (I). Data are from 20 fields from 5 mice. *p<0.01. (G) The expression of CD69 on electronically gated lamina propria CD4 T cells from control (green line) and DSS-treated (red line) WT mice. Background staining is shown as hatched areas.</p

DSS-induced colitis was restored by adoptive transfer of wild-type (WT) CD4 T cells into CD69 KO mice.

<p>(A) The survival rates of WT, CD69 KO mice without cell transfer, and CD69-KO mice that received WT CD4 T cells (WT-CD4T), CD69 KO CD4 T cells (KO-CD4T), or WT neutrophils (WT-Neutro) during DSS treatment. Survival was recorded daily (n = 5 per group). (B, C) Changes in the disease activity index (B) and body weight (%) (C) over the course of DSS treatment. The data are presented as the means (SD) (n = 5 per group). *p<0.05. (D) Histological sections of inflamed colons. Colons were taken on day 8 from mice receiving DSS in their drinking water. Sections were fixed and stained with hematoxylin and eosin (original magnification 40×). Two independent experiments were performed with similar results.</p

Inhibition of dextran sulphate sodium (DSS)-induced acute colitis in CD69-deficient (CD69 KO) mice.

<p>Acute colitis was induced by giving animals 4% DSS in drinking water for 7 days, followed by normal drinking water. (A) The survival rates of CD69 KO and wild-type (WT) mice after the initiation of during DSS-induced acute colitis. The survival was recorded daily (n = 10 per group). (B, C) Changes in the disease activity index (B) and body weight (%) (C) over the course of DSS treatment in CD69 KO and WT mice. The data are presented as the means (SD) (n = 10 per group). *p<0.01. Three independent experiments were performed with similar results.</p

Crucial Role for CD69 in the Pathogenesis of Dextran Sulphate Sodium-Induced Colitis

<div><p>CD69 is a membrane molecule transiently expressed on activated lymphocytes, and its selective expression in inflammatory infiltrates suggests that it plays a role in the pathogenesis of inflammatory diseases. In this study, we used CD69-deficient (CD69 KO) mice to assess the role of CD69 in the pathogenesis of dextran sulphate sodium (DSS)-induced acute and chronic colitis. The severity of colitis was assessed by the survival rate, clinical signs, colon length, histological examination and the expression of cytokines and chemokines in the large intestines. Both acute and chronic colitis were attenuated in the CD69 KO mice, as reflected by the lower lethality, weight loss, clinical signs, and improved histological findings. CD69⁺ cells infiltrated extensively into the inflamed mucosa of the colon in WT mice after DSS treatment. Experiments with the transfer of WT CD4 T cells into CD69 KO mice restored the induction of colitis. The administration of an anti-CD69 antibody also inhibited the induction of the DSS-induced colitis. These results indicate that CD69 expressed on CD4 T cells plays an important role in the pathogenesis of DSS-induced acute and chronic colitis, and that CD69 could be a possible therapeutic target for colitis.</p></div

(A) Enforced expression of Noxa-induced cell death in effector Th2 cells after cytokine depletion

Effector Th2 cells infected with a –containing retrovirus were cultured in vitro for 24 h without cytokines. hNGFR profiles (left) and annexin V staining profiles of the electronically gated hNGFR (gate #2) and hNGFR (gate #1) populations are shown. Three independent experiments were performed with similar results. (B) KJ1 effector Th2 cells infected with –containing retrovirus were transferred into BALB/c mice. 5 wk later, memory Th2 cell generation was determined by KJ1/EGFP expression. Expression of EGFP in pretransferred effector Th2 cells (top left) and a typical KJ1/GFP profile of freshly prepared memory Th2 cells (top right) are shown. In the bottom panels, the percentages of KJ1 cells and GFP Noxa-overexpressing cells and the mean fluorescence intensity of the GFP cells are shown with standard deviations ( = 4). The experiments were performed twice with similar results. (C) The effector Th2 cells from /, /, and / mice (Ly5.2) were transferred into Ly5.1 host mice, and the number of Ly5.2 memory Th2 cells was determined. A typical staining pattern of CD4/Ly5.2 (top) and the percentages of Ly5.2 cells among CD4 T cells are shown with standard deviations ( = 5; bottom). Three independent experiments were performed with similar results. (D) Deletion of the gene enhanced the generation of memory Th2 cells. In vitro–generated effector Th2 cells (Ly5.2) were transferred into Ly5.1 host mice. 5 wk after cell transfer, the number of Ly5.2 memory Th2 cells was determined. A representative CD4/Ly5.2 profile (left) and the mean values with standard deviations ( = 5; right) are shown. The experiments were performed twice with similar results.<p><b>Copyright information:</b></p><p>Taken from "Bmi1 regulates memory CD4 T cell survival via repression of the gene"</p><p></p><p>The Journal of Experimental Medicine 2008;205(5):1109-1120.</p><p>Published online 12 May 2008</p><p>PMCID:PMC2373843.</p><p></p

(A) mRNA expression of and p53-related proapoptotic genes in effector Th2 cells was determined by quantitative RT-PCR

The relative intensity (/HPRT; mean of three samples) is shown with standard deviations. Three independent experiments were performed with similar results. (B) mRNA expression of , p53-related proapoptotic genes, and antiapoptotic genes in memory Th2 cells was analyzed. The relative intensity (/HPRT; mean of three samples) is shown with standard deviations. Two independent experiments were performed with similar results. (C) Effects on p16 and p19 deficiency on the memory Th2 cell generation. The effector Th2 cells from the indicated mice (Ly5.2 background) were transferred into Ly5.1 host mice. 5 wk after cell transfer, the number of Ly5.2 memory Th2 cells was determined. The mean values are shown with standard deviations ( = 5; right). The experiments were performed twice with similar results. (D) mRNA levels of proapoptotic genes in // effector Th2 cells were determined by quantitative RT-PCR. The relative intensity (/HPRT; mean of three samples) is shown with standard deviations. The experiments were performed twice with similar results.<p><b>Copyright information:</b></p><p>Taken from "Bmi1 regulates memory CD4 T cell survival via repression of the gene"</p><p></p><p>The Journal of Experimental Medicine 2008;205(5):1109-1120.</p><p>Published online 12 May 2008</p><p>PMCID:PMC2373843.</p><p></p