Natural regulatory T cells from patients with end-stage renal disease can be used for large-scale generation of highly suppressive alloantigen-specific Tregs

Abstract

Natural occurring regulatory T cells (nTregs) have the potential to offer a targeted approach of immunosuppression and are the cell type of interest for inducing tolerance in kidney transplantation. End-stage renal disease (ESRD) profoundly affects the composition and function of circulating T cells but little is known with respect to how nTreg potential is affected. To address this, nTregs of patients with ESRD (on dialysis or not) and healthy individuals were isolated, expanded using allogeneic mature monocyte-derived dendritic cells followed by anti-CD3/anti-CD28-coated beads and the different nTregs were extensively characterized by the demethylation status of the Treg-specific demethylated region within FOXP3 and expression of typical nTreg markers. Additionally, the suppressive capacity as well as cytokine producing cells were analyzed for allogeneic mature monocyte-derived dendritic cell-expanded nTregs. Compared to age- and gender-matched healthy individuals, similar frequencies of nTregs were present within the circulation of patients with ESRD either on dialysis or not. The isolated nTregs could be equally well or even better expanded using allogeneic mature monocyte-derived dendritic cells and extensive characterization did not reveal significant differences. The demethylation status of the Treg-specific demethylated region was maintained or even further promoted as was the expression of markers characteristic for nTregs. Moreover, suppressive capacity and the cytokine profile of allogeneic mature monocyte-derived dendritic cell-expanded nTregs was similar to that of healthy individuals. Thus, circulating nTregs of patients with ESRD can effectively be expanded to stable allo antigen-specific nTregs with potential clinical applicability.

Regulatory T cells in renal disease

“…in the ONE Study, http://www.onestudy.org). Early reports show the feasibility of expanding nTregs with allogeneic DCs and support the safety of cell therapy with nTregs . Table lists some of the differences between the use of human and mouse Tregs.…”

“…Clearly, unlike autoimmune renal disease, the timing of transplantation allows the infusion of Tregs prior to allogenic stimulation at the time of surgery. After nTreg isolation, both nonantigen‐specific and allogeneic stimulation and expansion protocols are being employed, including the use of belatacept (CTLA 4‐Fc) ex vivo . These protocols have been generating cells that maintain their phenotype including demethylation of the FOXP3 TSDR .…”

“…After nTreg isolation, both nonantigen‐specific and allogeneic stimulation and expansion protocols are being employed, including the use of belatacept (CTLA 4‐Fc) ex vivo . These protocols have been generating cells that maintain their phenotype including demethylation of the FOXP3 TSDR . Furthermore, Tr1 cells can also be generated from patients with end‐stage kidney disease who are on dialysis …”

“…107,122 These protocols have been generating cells that maintain their phenotype including demethylation of the FOXP3 TSDR. 107 Furthermore, Tr1 cells can also be generated from patients with end-stage kidney disease who are on dialysis. 123 A further dimension in the role of Tregs in transplantation is the effect of current and new immunosuppressive agents on Tregs and their interactions with Tregs.…”

Treating inflammation to improve outcome in dialysis patients

“…• Nell'ambito della terapia cellulare, le cellule T regolatorie (Treg) (27) e le cellule mesenchimali (28) sono studiate per le possibili implicazioni nell'indurre la tolleranza immunitaria nei pazienti portatori di trapianto di rene, con la loro possibile azione antinfiammatoria nei pazienti in emodialisi cronica.…”