Limits...
Foxp3+ Treg expanded from patients with established diabetes reduce Helios expression while retaining normal function compared to healthy individuals.

Du W, Shen YW, Lee WH, Wang D, Paz S, Kandeel F, Liu CP - PLoS ONE (2013)

Bottom Line: After isolation, Treg from both T1D patients and healthy subjects were successfully expanded with high purity.Importantly, expanded Treg from both subject groups were able to suppress autologous or allogeneic CD8(+) effector T cells equally well.Thus, based on the positive outcomes, these potent expanded Treg from diabetic human patients may be useful in treating T1D or preventing islet graft rejection.

View Article: PubMed Central - PubMed

Affiliation: Department of Diabetes and Metabolic Diseases Research, Beckman Research Institute, City of Hope, Duarte, California, United States of America.

ABSTRACT
Foxp3(+) regulatory T cells (Treg) play a crucial role in regulating immune tolerance. The use of Treg to restore immune tolerance is considered an attractive novel approach to inhibit autoimmune disease, including type 1 diabetes (T1D), and to prevent rejection of organ transplants. In view of the goal of developing autologous Treg-based cell therapy for patients with long-term (>15 years) T1D, it will be necessary to expand a sufficient amount of functional Treg in vitro in order to study and compare Treg from T1D patients and healthy subjects. Our results have demonstrated that there is a comparable frequency of Treg in the peripheral blood lymphocytes (PBLs) of patients with long-term T1D relative to those in healthy subjects; however, Th1 cells, but not Th17 cells, were increased in the T1D patients. Further, more Treg in PBLs from T1D patients than from healthy subjects expressed the CD45RO(+) memory cell phenotype, suggesting they were antigen-experienced cells. After isolation, Treg from both T1D patients and healthy subjects were successfully expanded with high purity. Although there was no difference in Helios expression on Treg in PBLs, in vitro expansion led to fewer Helios-expressing Treg from T1D patients than healthy subjects. While more Th1-like Treg expressing IFN-γ or TNF-α were found in the PBLs of T1D patients than healthy controls, there was no such difference in the expanded Treg. Importantly, expanded Treg from both subject groups were able to suppress autologous or allogeneic CD8(+) effector T cells equally well. Our findings demonstrate that a large number of ex vivo expanded functional Treg can be obtained from long-term T1D patients, although fewer expanded Treg expressed a high level of Helios. Thus, based on the positive outcomes, these potent expanded Treg from diabetic human patients may be useful in treating T1D or preventing islet graft rejection.

Show MeSH

Related in: MedlinePlus

Increased IFN-γ or TNF-α-producing Th1 cells in T1D patients.PBMCs were stained with indicated antibodies following activation with PMA/ionomycin for 5 h in vitro and analyzed by FACS. (A) Percentage of IFN-γ+, TNF-α+,or IL17+ cells present in CD4+ T cells of PBLs from T1D patients or healthy controls. (B) The ratios of Th1/Treg, Th1/Th17, and Th17/Treg in CD4+ T cells from PBLs of diabetic patients or healthy controls.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC3569420&req=5

pone-0056209-g006: Increased IFN-γ or TNF-α-producing Th1 cells in T1D patients.PBMCs were stained with indicated antibodies following activation with PMA/ionomycin for 5 h in vitro and analyzed by FACS. (A) Percentage of IFN-γ+, TNF-α+,or IL17+ cells present in CD4+ T cells of PBLs from T1D patients or healthy controls. (B) The ratios of Th1/Treg, Th1/Th17, and Th17/Treg in CD4+ T cells from PBLs of diabetic patients or healthy controls.

Mentions: Although T1D is a T-cell-mediated disease, the specific pathogenic mechanisms leading to T1D remain largely unclear. In particular, further studies are needed to elucidate the relative roles of Th1 and Th17 cells versus Treg during the onset and further development of T1D in human patients [37]. One hypothesis suggests that imbalanced pro-inflammatory T cell subsets (including Th1 and Th17 cells) versus Treg in T1D patients may contribute to T1D pathogenesis. To begin addressing this issue, we performed intracellular staining analyses of IFN-γ, TNF-α, IL17, or Foxp3 on PBLs of T1D and healthy subjects. The results showed an elevated IFN-γ (p = 0.02) and TNF-α (p = 0.03) expression in CD4+ T cells in PBLs of T1D subjects than those of healthy controls (Fig. 6A). In comparison, only a small percentage of IL17-expressing CD4+ cells were present in both T1D subjects and healthy controls, and there was no difference between the two groups (Fig. 6A). Further analyses showed that the observed difference in cell phenotype is independent of the age of subjects in the two cohorts (data not shown). Therefore, a significantly increased Th1 but not Th17 cell population is present in the PBLs of T1D patients compared to those of healthy controls.


Foxp3+ Treg expanded from patients with established diabetes reduce Helios expression while retaining normal function compared to healthy individuals.

Du W, Shen YW, Lee WH, Wang D, Paz S, Kandeel F, Liu CP - PLoS ONE (2013)

Increased IFN-γ or TNF-α-producing Th1 cells in T1D patients.PBMCs were stained with indicated antibodies following activation with PMA/ionomycin for 5 h in vitro and analyzed by FACS. (A) Percentage of IFN-γ+, TNF-α+,or IL17+ cells present in CD4+ T cells of PBLs from T1D patients or healthy controls. (B) The ratios of Th1/Treg, Th1/Th17, and Th17/Treg in CD4+ T cells from PBLs of diabetic patients or healthy controls.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3569420&req=5

pone-0056209-g006: Increased IFN-γ or TNF-α-producing Th1 cells in T1D patients.PBMCs were stained with indicated antibodies following activation with PMA/ionomycin for 5 h in vitro and analyzed by FACS. (A) Percentage of IFN-γ+, TNF-α+,or IL17+ cells present in CD4+ T cells of PBLs from T1D patients or healthy controls. (B) The ratios of Th1/Treg, Th1/Th17, and Th17/Treg in CD4+ T cells from PBLs of diabetic patients or healthy controls.
Mentions: Although T1D is a T-cell-mediated disease, the specific pathogenic mechanisms leading to T1D remain largely unclear. In particular, further studies are needed to elucidate the relative roles of Th1 and Th17 cells versus Treg during the onset and further development of T1D in human patients [37]. One hypothesis suggests that imbalanced pro-inflammatory T cell subsets (including Th1 and Th17 cells) versus Treg in T1D patients may contribute to T1D pathogenesis. To begin addressing this issue, we performed intracellular staining analyses of IFN-γ, TNF-α, IL17, or Foxp3 on PBLs of T1D and healthy subjects. The results showed an elevated IFN-γ (p = 0.02) and TNF-α (p = 0.03) expression in CD4+ T cells in PBLs of T1D subjects than those of healthy controls (Fig. 6A). In comparison, only a small percentage of IL17-expressing CD4+ cells were present in both T1D subjects and healthy controls, and there was no difference between the two groups (Fig. 6A). Further analyses showed that the observed difference in cell phenotype is independent of the age of subjects in the two cohorts (data not shown). Therefore, a significantly increased Th1 but not Th17 cell population is present in the PBLs of T1D patients compared to those of healthy controls.

Bottom Line: After isolation, Treg from both T1D patients and healthy subjects were successfully expanded with high purity.Importantly, expanded Treg from both subject groups were able to suppress autologous or allogeneic CD8(+) effector T cells equally well.Thus, based on the positive outcomes, these potent expanded Treg from diabetic human patients may be useful in treating T1D or preventing islet graft rejection.

View Article: PubMed Central - PubMed

Affiliation: Department of Diabetes and Metabolic Diseases Research, Beckman Research Institute, City of Hope, Duarte, California, United States of America.

ABSTRACT
Foxp3(+) regulatory T cells (Treg) play a crucial role in regulating immune tolerance. The use of Treg to restore immune tolerance is considered an attractive novel approach to inhibit autoimmune disease, including type 1 diabetes (T1D), and to prevent rejection of organ transplants. In view of the goal of developing autologous Treg-based cell therapy for patients with long-term (>15 years) T1D, it will be necessary to expand a sufficient amount of functional Treg in vitro in order to study and compare Treg from T1D patients and healthy subjects. Our results have demonstrated that there is a comparable frequency of Treg in the peripheral blood lymphocytes (PBLs) of patients with long-term T1D relative to those in healthy subjects; however, Th1 cells, but not Th17 cells, were increased in the T1D patients. Further, more Treg in PBLs from T1D patients than from healthy subjects expressed the CD45RO(+) memory cell phenotype, suggesting they were antigen-experienced cells. After isolation, Treg from both T1D patients and healthy subjects were successfully expanded with high purity. Although there was no difference in Helios expression on Treg in PBLs, in vitro expansion led to fewer Helios-expressing Treg from T1D patients than healthy subjects. While more Th1-like Treg expressing IFN-γ or TNF-α were found in the PBLs of T1D patients than healthy controls, there was no such difference in the expanded Treg. Importantly, expanded Treg from both subject groups were able to suppress autologous or allogeneic CD8(+) effector T cells equally well. Our findings demonstrate that a large number of ex vivo expanded functional Treg can be obtained from long-term T1D patients, although fewer expanded Treg expressed a high level of Helios. Thus, based on the positive outcomes, these potent expanded Treg from diabetic human patients may be useful in treating T1D or preventing islet graft rejection.

Show MeSH
Related in: MedlinePlus