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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

LPL Tregs from IBD patients are suppressive in vitro.CD25+, CD127-, CD4+ LPL Tregs homogenized from the surgically resected colons of patients with or without IBD were sorted and added at a 1:2 ratio to CFSE-labeled splenocytes from a single donor without IBD or other inflammatory or malignant conditions. As a control, autologous Tregs were similarly sorted from the splenocytes as well. Cells were cocultured in the presence of soluble anti-CD3 (OKT3) for 4 days and then CFSE dilution by CD4+ splenocytes was quantified on a flow cytometer. Representative histograms of CFSE dilution by these CD4+ splenocytes in the absence or presence of Tregs from different donors are shown (a). The percent that CD4+ splenocyte CFSE dilution was diminished by the presence of LPL Tregs from the indicated donors is shown (b). Each dot is a unique patient, except “Non-IBD Spleen”, for which Tregs from the same spleen donor were isolated de novo on each of three different days to serve as a control. CD = Crohn’s disease. UC = ulcerative colitis. LPL = lamina propria lymphocytes.
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pone.0136485.g001: LPL Tregs from IBD patients are suppressive in vitro.CD25+, CD127-, CD4+ LPL Tregs homogenized from the surgically resected colons of patients with or without IBD were sorted and added at a 1:2 ratio to CFSE-labeled splenocytes from a single donor without IBD or other inflammatory or malignant conditions. As a control, autologous Tregs were similarly sorted from the splenocytes as well. Cells were cocultured in the presence of soluble anti-CD3 (OKT3) for 4 days and then CFSE dilution by CD4+ splenocytes was quantified on a flow cytometer. Representative histograms of CFSE dilution by these CD4+ splenocytes in the absence or presence of Tregs from different donors are shown (a). The percent that CD4+ splenocyte CFSE dilution was diminished by the presence of LPL Tregs from the indicated donors is shown (b). Each dot is a unique patient, except “Non-IBD Spleen”, for which Tregs from the same spleen donor were isolated de novo on each of three different days to serve as a control. CD = Crohn’s disease. UC = ulcerative colitis. LPL = lamina propria lymphocytes.

Mentions: To confirm previously published data[9] that functional Tregs are present in the intestinal mucosa of IBD patients, we were able to sort sufficient numbers of CD25+/CD127- CD4+ T cells from the homogenized, surgically resected intestinal LPL of three UC patients, three Crohn’s patients, and one patient without IBD to perform in vitro suppression assays. As a consistent source of large numbers of responder T cells for these assays, the splenocytes of a single distal pancreatectomy recipient with no autoimmunity or cancer were used for all experiments. Proliferation of CD4+ (Fig 1a and 1b) and CD8+ (data not shown) splenocytes was comparably inhibited by the presence of Tregs, regardless of whether the Tregs were from patients with or without IBD.


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)

LPL Tregs from IBD patients are suppressive in vitro.CD25+, CD127-, CD4+ LPL Tregs homogenized from the surgically resected colons of patients with or without IBD were sorted and added at a 1:2 ratio to CFSE-labeled splenocytes from a single donor without IBD or other inflammatory or malignant conditions. As a control, autologous Tregs were similarly sorted from the splenocytes as well. Cells were cocultured in the presence of soluble anti-CD3 (OKT3) for 4 days and then CFSE dilution by CD4+ splenocytes was quantified on a flow cytometer. Representative histograms of CFSE dilution by these CD4+ splenocytes in the absence or presence of Tregs from different donors are shown (a). The percent that CD4+ splenocyte CFSE dilution was diminished by the presence of LPL Tregs from the indicated donors is shown (b). Each dot is a unique patient, except “Non-IBD Spleen”, for which Tregs from the same spleen donor were isolated de novo on each of three different days to serve as a control. CD = Crohn’s disease. UC = ulcerative colitis. LPL = lamina propria lymphocytes.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0136485.g001: LPL Tregs from IBD patients are suppressive in vitro.CD25+, CD127-, CD4+ LPL Tregs homogenized from the surgically resected colons of patients with or without IBD were sorted and added at a 1:2 ratio to CFSE-labeled splenocytes from a single donor without IBD or other inflammatory or malignant conditions. As a control, autologous Tregs were similarly sorted from the splenocytes as well. Cells were cocultured in the presence of soluble anti-CD3 (OKT3) for 4 days and then CFSE dilution by CD4+ splenocytes was quantified on a flow cytometer. Representative histograms of CFSE dilution by these CD4+ splenocytes in the absence or presence of Tregs from different donors are shown (a). The percent that CD4+ splenocyte CFSE dilution was diminished by the presence of LPL Tregs from the indicated donors is shown (b). Each dot is a unique patient, except “Non-IBD Spleen”, for which Tregs from the same spleen donor were isolated de novo on each of three different days to serve as a control. CD = Crohn’s disease. UC = ulcerative colitis. LPL = lamina propria lymphocytes.
Mentions: To confirm previously published data[9] that functional Tregs are present in the intestinal mucosa of IBD patients, we were able to sort sufficient numbers of CD25+/CD127- CD4+ T cells from the homogenized, surgically resected intestinal LPL of three UC patients, three Crohn’s patients, and one patient without IBD to perform in vitro suppression assays. As a consistent source of large numbers of responder T cells for these assays, the splenocytes of a single distal pancreatectomy recipient with no autoimmunity or cancer were used for all experiments. Proliferation of CD4+ (Fig 1a and 1b) and CD8+ (data not shown) splenocytes was comparably inhibited by the presence of Tregs, regardless of whether the Tregs were from patients with or without IBD.

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