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. 2001 Jul;49(1):35–41. doi: 10.1136/gut.49.1.35

Decreased Bax expression by mucosal T cells favours resistance to apoptosis in Crohn's disease

J Itoh 1, C de la Motte 1, S Strong 1, A Levine 1, C Fiocchi 1
PMCID: PMC1728348  PMID: 11413108

Abstract

BACKGROUND—Activated T cells are more susceptible to apoptosis than resting T cells. As intestinal T cells normally exhibit a higher state of activation, increased apoptosis may be necessary to maintain immune homeostasis in the specialised microenvironment of the mucosa. On the other hand, in Crohn's disease (CD) mucosal T cells are resistant to apoptosis, suggesting abnormal regulation of cell death mechanisms.
AIMS—To investigate differences in expression of anti- and proapoptotic Bcl-2 family proteins, key regulators of apoptosis, between circulating and mucosal T cells, and possible alterations in CD.
PATIENTS AND METHODS—Lamina propria T cells (LPT) were isolated from 10 control, seven CD, and eight ulcerative colitis (UC) patients, and peripheral blood T cells (PBT) from healthy volunteers. Purified T cells were stained intracellularly for Bcl-2, Bcl-xL, and Bax, and mean fluorescence intensity measured by flow cytometry.
RESULTS—Compared with PBT, the expression level of Bcl-2 and Bax, but not Bcl-xL, was significantly greater in LPT, resulting in lower Bcl-xL/Bax ratios. In PBT, Bax expression was highly and significantly correlated with both Bcl-2 and Bcl-xL, but correlation with Bcl-2 was absent in LPT. Bax expression in CD, but not UC, LPT was significantly lower than in control LPT, resulting in a significantly higher Bcl-xL/Bax ratio. The significant correlation of Bcl-xL to Bax was preserved in CD, but not UC, LPT.
CONCLUSIONS—Regulation of Bcl-2 family protein expression differs between circulating and mucosal T cells, probably underlying diverse survival potentials. In CD LPT, a low Bax expression and a high Bcl-xL/Bax ratio favour resistance to apoptosis and may contribute to the chronicity of inflammation.


Keywords: Bcl-2 family proteins; apoptosis; inflammatory bowel disease; T cells; Crohn's disease; ulcerative colitis

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Figure 1  .

Figure 1  

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Representative flow cytometric analysis of intracellular staining for Bcl-2, Bcl-xL, and Bax protein levels in normal peripheral blood T cells (PBT) and control lamina propria T cells (LPT).

Figure 2  .

Figure 2  

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Flow cytometric analysis of intracellular staining for Bcl-2, Bcl-xL, and Bax protein levels in normal peripheral blood T cells (PBT) and control lamina propria T cells (LPT). Results are mean (SEM) fluorescence intensity for eight PBT and 10 LPT isolates. **p<0.002, ***p<0.001.

Figure 3  .

Figure 3  

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Flow cytometric analysis of intracellular staining for Bcl-2, Bcl-xL, and Bax protein levels in normal unseparated and CD45RO enriched (+) peripheral blood T cells (PBT). Results are mean (SEM) fluorescence intensity for eight PBT isolates. No statistically significant differences were observed for Bcl-2, Bcl-xL, and Bax.

Figure 4  .

Figure 4  

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Correlation of anti- and proapoptotic Bcl-2 family protein levels in normal peripheral blood T cells (PBT) (top) and control lamina propria T cells (LPT) (bottom). Individual data points represent mean fluorescence intensity of intracellular staining measured by flow cytometric analysis.

Figure 5  .

Figure 5  

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Representative flow cytometric analysis of intracellular staining for Bcl-2, Bcl-xL, and Bax protein levels in control, Crohn's disease (CD) and ulcerative colitis (UC) lamina propria T cells (LPT).

Figure 6  .

Figure 6  

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Flow cytometric analysis of intracellular staining for Bcl-2, Bcl-xL, and Bax protein levels in control, Crohn's disease (CD), and ulcerative colitis (UC) lamina propria T cells (LPT). Results are mean (SEM) fluorescence intensity for 10 control, seven CD, and eight UC isolates. **p<0.003, CD versus control and UC.

Figure 7  .

Figure 7  

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Correlation of proapoptotic Bax and antiapoptotic Bcl-xL protein levels in Crohn's disease (CD) and ulcerative colitis (UC) lamina propria T cells (LPT). Individual data points represent mean fluorescence intensity of intracellular staining measured by flow cytometric analysis.

Figure 8  .

Figure 8  

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Comparative correlation of proapoptotic Bax and antiapoptotic Bcl-xL in control and Crohn's disease (CD) lamina propria T cells (LPT). Individual data points represent mean fluorescence intensity of intracellular staining measured by flow cytometric analysis.

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