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Human Blood and Mucosal Regulatory T Cells Express Activation Markers and Inhibitory Receptors in Inflammatory Bowel Disease.

Lord JD, Shows DM, Chen J, Thirlby RC - PLoS ONE (2015)

Bottom Line: In all samples, a similar fraction of FOXP3+ cells expressed the "natural" Treg (nTreg) marker Helios, suggesting that, in IBD, these cells are not entirely "induced" Tregs (iTregs) derived from activated effector T cells.Greater expression of activation markers was also seen in both Helios+ and Helios- Tregs, relative to FOXP3- cells, in both IBD patients and controls, indicating that Tregs are effectively activated by antigen in IBD.Extensive immunophenotyping revealed that Helios+ and Helios- mucosal Tregs exist at a similar frequency, and have a similar expression of inhibitory molecules and activation markers in patients with IBD as in healthy controls.

View Article: PubMed Central - PubMed

Affiliation: Translational Research Program at the Benaroya Research Institute at Virginia Mason, Seattle Washington, United States of America.

ABSTRACT

Background: FOXP3+ regulatory T cells (Tregs) are critical for preventing intestinal inflammation. However, FOXP3+ T cells are paradoxically increased in the intestines of patients with the inflammatory bowel disease (IBD) ulcerative colitis (UC) or Crohn's disease (CD). We determined whether these FOXP3+ cells in IBD patients share or lack the phenotype of such cells from patients without IBD.

Methods: We quantified and characterized FOXP3+ Treg populations, as well as FOXP3- CD4+ T cells, in the lamina propria lymphocytes (LPL) of intestine surgically resected from patients with and without IBD, and in the blood of controls or Crohn's patients with or without disease activity.

Results: In all samples, a similar fraction of FOXP3+ cells expressed the "natural" Treg (nTreg) marker Helios, suggesting that, in IBD, these cells are not entirely "induced" Tregs (iTregs) derived from activated effector T cells. Helios+ and Helios- FOXP3+ T cells demonstrated similar expression of maturation markers, activation markers, and inhibitory molecules between IBD patients and controls, while FOXP3- cells paradoxically expressed more of the inhibitory receptors CD39, CTLA4, and PD-1 in inflamed mucosa. Greater expression of activation markers was also seen in both Helios+ and Helios- Tregs, relative to FOXP3- cells, in both IBD patients and controls, indicating that Tregs are effectively activated by antigen in IBD.

Conclusions: Extensive immunophenotyping revealed that Helios+ and Helios- mucosal Tregs exist at a similar frequency, and have a similar expression of inhibitory molecules and activation markers in patients with IBD as in healthy controls.

No MeSH data available.


Related in: MedlinePlus

Increased intestinal FOXP3+ T cells in IBD are not predominantly Helios-.Homegenized LPL (a,c) or PBMC (b,d) were thawed and stained according to Table 1, or with isotype control antibodies. Gating on CD3+, CD4+ T cells, the percent of colonic or ileal LPL expressing FOXP3 is shown according to whether tissue was inflamed or not (a). The percent of CD3+, CD4+ PBMC expressing FOXP3 is also shown for a separate cohort of Crohn’s patients, with or without symptoms, or for matched controls (b). Data for a and b was re-gated on CD3+, CD4+, FOXP3+ cells and the percent expressing Helios is shown as above (c, d). Each point represents a unique specimen. Bars indicate means and standard deviations. For c and d, each data point is the mean of three independent assays.
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pone.0136485.g003: Increased intestinal FOXP3+ T cells in IBD are not predominantly Helios-.Homegenized LPL (a,c) or PBMC (b,d) were thawed and stained according to Table 1, or with isotype control antibodies. Gating on CD3+, CD4+ T cells, the percent of colonic or ileal LPL expressing FOXP3 is shown according to whether tissue was inflamed or not (a). The percent of CD3+, CD4+ PBMC expressing FOXP3 is also shown for a separate cohort of Crohn’s patients, with or without symptoms, or for matched controls (b). Data for a and b was re-gated on CD3+, CD4+, FOXP3+ cells and the percent expressing Helios is shown as above (c, d). Each point represents a unique specimen. Bars indicate means and standard deviations. For c and d, each data point is the mean of three independent assays.

Mentions: We performed cell-surface and intracellular staining on thawed LPL or PBMC from IBD patients versus controls to identify CD4+ FOXP3+ Tregs, and divide them into Helios+ nTregs and Helios- iTregs. As has been previously described, a greater fraction of the CD4+ T cell population was FOXP3+ in the colons of IBD patients than controls (p = 0.0307). This trend was seen regardless of whether or not the mucosa from which LPL were derived was grossly inflamed (Fig 3a), but was not seen in the peripheral blood of Crohn’s patients relative to controls, regardless of symptomatic disease activity (Fig 3b). A lower fraction of FOXP3+ T cells expressed the nTreg marker Helios in LPL (Fig 3c) than PBMC (Fig 3d) in all cohorts (p<0.0001), but the percent of FOXP3+ T cells expressing Helios was no different in the LPL of patients with or without IBD, regardless of inflammation (Fig 3c). Thus the increase in FOXP3+ T cells seen in IBD colon must be similar in both Helios+ and Helios- subpopulations. A slightly higher frequency of Tregs expressed Helios in the blood of symptomatic than asymptomatic Crohn’s patients, but not significantly, and neither cohort differed substantially from healthy controls (Fig 3d).


Human Blood and Mucosal Regulatory T Cells Express Activation Markers and Inhibitory Receptors in Inflammatory Bowel Disease.

Lord JD, Shows DM, Chen J, Thirlby RC - PLoS ONE (2015)

Increased intestinal FOXP3+ T cells in IBD are not predominantly Helios-.Homegenized LPL (a,c) or PBMC (b,d) were thawed and stained according to Table 1, or with isotype control antibodies. Gating on CD3+, CD4+ T cells, the percent of colonic or ileal LPL expressing FOXP3 is shown according to whether tissue was inflamed or not (a). The percent of CD3+, CD4+ PBMC expressing FOXP3 is also shown for a separate cohort of Crohn’s patients, with or without symptoms, or for matched controls (b). Data for a and b was re-gated on CD3+, CD4+, FOXP3+ cells and the percent expressing Helios is shown as above (c, d). Each point represents a unique specimen. Bars indicate means and standard deviations. For c and d, each data point is the mean of three independent assays.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4548948&req=5

pone.0136485.g003: Increased intestinal FOXP3+ T cells in IBD are not predominantly Helios-.Homegenized LPL (a,c) or PBMC (b,d) were thawed and stained according to Table 1, or with isotype control antibodies. Gating on CD3+, CD4+ T cells, the percent of colonic or ileal LPL expressing FOXP3 is shown according to whether tissue was inflamed or not (a). The percent of CD3+, CD4+ PBMC expressing FOXP3 is also shown for a separate cohort of Crohn’s patients, with or without symptoms, or for matched controls (b). Data for a and b was re-gated on CD3+, CD4+, FOXP3+ cells and the percent expressing Helios is shown as above (c, d). Each point represents a unique specimen. Bars indicate means and standard deviations. For c and d, each data point is the mean of three independent assays.
Mentions: We performed cell-surface and intracellular staining on thawed LPL or PBMC from IBD patients versus controls to identify CD4+ FOXP3+ Tregs, and divide them into Helios+ nTregs and Helios- iTregs. As has been previously described, a greater fraction of the CD4+ T cell population was FOXP3+ in the colons of IBD patients than controls (p = 0.0307). This trend was seen regardless of whether or not the mucosa from which LPL were derived was grossly inflamed (Fig 3a), but was not seen in the peripheral blood of Crohn’s patients relative to controls, regardless of symptomatic disease activity (Fig 3b). A lower fraction of FOXP3+ T cells expressed the nTreg marker Helios in LPL (Fig 3c) than PBMC (Fig 3d) in all cohorts (p<0.0001), but the percent of FOXP3+ T cells expressing Helios was no different in the LPL of patients with or without IBD, regardless of inflammation (Fig 3c). Thus the increase in FOXP3+ T cells seen in IBD colon must be similar in both Helios+ and Helios- subpopulations. A slightly higher frequency of Tregs expressed Helios in the blood of symptomatic than asymptomatic Crohn’s patients, but not significantly, and neither cohort differed substantially from healthy controls (Fig 3d).

Bottom Line: In all samples, a similar fraction of FOXP3+ cells expressed the "natural" Treg (nTreg) marker Helios, suggesting that, in IBD, these cells are not entirely "induced" Tregs (iTregs) derived from activated effector T cells.Greater expression of activation markers was also seen in both Helios+ and Helios- Tregs, relative to FOXP3- cells, in both IBD patients and controls, indicating that Tregs are effectively activated by antigen in IBD.Extensive immunophenotyping revealed that Helios+ and Helios- mucosal Tregs exist at a similar frequency, and have a similar expression of inhibitory molecules and activation markers in patients with IBD as in healthy controls.

View Article: PubMed Central - PubMed

Affiliation: Translational Research Program at the Benaroya Research Institute at Virginia Mason, Seattle Washington, United States of America.

ABSTRACT

Background: FOXP3+ regulatory T cells (Tregs) are critical for preventing intestinal inflammation. However, FOXP3+ T cells are paradoxically increased in the intestines of patients with the inflammatory bowel disease (IBD) ulcerative colitis (UC) or Crohn's disease (CD). We determined whether these FOXP3+ cells in IBD patients share or lack the phenotype of such cells from patients without IBD.

Methods: We quantified and characterized FOXP3+ Treg populations, as well as FOXP3- CD4+ T cells, in the lamina propria lymphocytes (LPL) of intestine surgically resected from patients with and without IBD, and in the blood of controls or Crohn's patients with or without disease activity.

Results: In all samples, a similar fraction of FOXP3+ cells expressed the "natural" Treg (nTreg) marker Helios, suggesting that, in IBD, these cells are not entirely "induced" Tregs (iTregs) derived from activated effector T cells. Helios+ and Helios- FOXP3+ T cells demonstrated similar expression of maturation markers, activation markers, and inhibitory molecules between IBD patients and controls, while FOXP3- cells paradoxically expressed more of the inhibitory receptors CD39, CTLA4, and PD-1 in inflamed mucosa. Greater expression of activation markers was also seen in both Helios+ and Helios- Tregs, relative to FOXP3- cells, in both IBD patients and controls, indicating that Tregs are effectively activated by antigen in IBD.

Conclusions: Extensive immunophenotyping revealed that Helios+ and Helios- mucosal Tregs exist at a similar frequency, and have a similar expression of inhibitory molecules and activation markers in patients with IBD as in healthy controls.

No MeSH data available.


Related in: MedlinePlus