Foxp3+ Treg Expanded from Patients with Established Diabetes Reduce Helios Expression while Retaining Normal Function Compared to Healthy Individuals

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.

Divergent effects of type-I interferons on regulatory T cells

“…IFN-g-producing Foxp3+ cells, or so-called Th1-like cells, can be polarized in vitro from conventional Treg upon the sequential exposure to IFN-g and then to IL-12, as demonstrated in the mouse system [36] and as we have confirmed using human Treg [9]. Th1-like Treg have been observed in a variety of inflammatory settings dominated by type-1 responses, such as graft-versus-host disease [37], viral infection [38], parasite infection [39], MS [40] and diabetes [41,42]. We have recently described that Th1-like Treg accumulate in the inflamed liver tissue of patients with chronic HCV infection, in line with the local abundance of IL-12 and IFN-g.…”

Soluble OX40L and JAG1 Induce Selective Proliferation of Functional Regulatory T-Cells Independent of canonical TCR signaling

“…TIGIT + subset of Tregs have been shown to predominantly inhibit Th1 and Th17 cells without affecting Th2 cells40. Helios, an Ikoras transcription factor family member, has also been reported to be associated with Treg functions41 and suppression of autoimmune diabetes42. Increased expression of these suppressive markers in OX40L-JAG1 expanded Tregs could help sustain their suppressive functions.…”

Low expression of CD39+/CD45RA+ on regulatory T cells (Treg) cells in type 1 diabetic children in contrast to high expression of CD101+/CD129+ on Treg cells in children with coeliac disease

“…In terms of additional markers for delineating cells of T reg lineage, CD39 and CD45RA have been included as possible markers for detection of T reg cells. Thus far, it has been shown that T reg cells, defined as CD4 + FoxP3 + , express more CD45RO but less CD45RA in T1D patients compared to healthy controls . Also in fulminant T1D patients, the frequency of CD45RA − FoxP3 high activated T reg cells is shown to be significantly lower compared to patients with type 1A diabetes .…”