Third party, umbilical cord blood derived regulatory T-cells for prevention of graft versus host disease in allogeneic hematopoietic stem cell transplantation: feasibility, safety and immune reconstitution

Joshua N Kellner, Eveline M Delemarre, Eric Yvon, Stefan Nierkens, Jaap J Boelens, Ian McNiece, Amanda Olson, Yago Nieto, Stefan Ciurea, Uday Popat, Sairah Ahmed, Richard Champlin, Jennifer Ramos, Mitsutaka Nishimoto, Hongbing Ma, Zeng Ke, Peter Thall, Joseph D Khoury, Robert Negrin, Borje Andersson, Simrit Parmar, Joshua N Kellner, Eveline M Delemarre, Eric Yvon, Stefan Nierkens, Jaap J Boelens, Ian McNiece, Amanda Olson, Yago Nieto, Stefan Ciurea, Uday Popat, Sairah Ahmed, Richard Champlin, Jennifer Ramos, Mitsutaka Nishimoto, Hongbing Ma, Zeng Ke, Peter Thall, Joseph D Khoury, Robert Negrin, Borje Andersson, Simrit Parmar

Abstract

Incubation of umbilical cord blood (UCB) derived regulatory T-cells (Tregs) with fucosyltransferase enzyme improves their ability to home to the target tissue to prevent graft vs. host disease (GVHD). We report results of 5 patients (Double UCB Transplant, n=2; Peripheral Blood Matched Unrelated Donor Transplant, n=3) who received UCB-Tregs (Dose level = 1×106/kg), infused one day prior to the donor graft. All patients received their designated UCB-Treg dose without any infusion reaction. The ratio of conventional T-cells in donor graft was at least 10 times higher than infused UCB-Tregs (ratio range, 12-356). All patients engrafted at median of 13 days (range, 8-17 days). One patient died due to brain hemorrhage on day 45. A bi-modal increase of plasma IL-10 level occurred on day 7 and day 21 and notably, plasma IL-2 level dropped significantly in all patients at Day 7. All evaluable patients developed ≥grade II acute GVHD and at 1 year follow up, all were alive and without evidence of disease relapse. No increase in the chronic GVHD biomarkers (REG3a and Elafin) was observed at day 7. At the time of last follow up, all evaluable patients were off immune-suppression. Stage 2 of this clinical trial examining UCB-Treg at dose level= 1×107/kg is currently underway.

Keywords: cell therapy; graft versus host disease; regulatory T cells; stem cell transplantation; umbilical cord blood.

Conflict of interest statement

CONFLICTS OF INTEREST SP is a scientific founder and serves on the board of director and chief medical officer of Cellenkos, MDACC owned startup and has a license agreement with Cellenkos. RN is a consultant for Cellenkos Inc. The remaining authors declare no conflicts of interest.

Figures

Figure 1. Characterization of clinical ex-vivo expanded…
Figure 1. Characterization of clinical ex-vivo expanded CB Tregs
(A) Representative flow cytometry analysis of CB Tregs. Top row is Day 0 isolation of CD25 cells. Bottom row is Day 14 expanded Tregs. Far right panels: CLA expression at Day 14 Pre- (top) and Post- (bottom) fucosylation. (B) Total expanded viable cells counted at each time point in culture. Results are mean ± SEM. (C) Representative flow plots of Treg:Tcon suppression assay from ex vivo expanded CB Tregs.
Figure 2. Patient outcomes
Figure 2. Patient outcomes
(A) Cumulative Incidence: ANC (Absolute Neutrophil Count) and Platelet engraftment. (B) Overall Survival. (C) FOXP3 immunohistochemistry showing Treg infiltration in tissue samples with graft-versus-host disease (GVHD). (A) Treg cells in a duodenal biopsy showing clustering around areas of epithelial distortion by GVHD (inset; thin arrow). Higher power shows exocytosis of Tregs into glandular epithelium (thick arrow). (B) Similar findings were seen in a colonic biopsy. [A: 40x (inset 20x); B: 40x].
Figure 3. Hematopoietic analysis of patient peripheral…
Figure 3. Hematopoietic analysis of patient peripheral blood
(A) Flow cytometric analysis of number of Tregs (CD4+CD25+CD127-). (B) Flow cytometric analysis of percent IL-10 secreting cells in PB. (C) Plasma analysis of IL-10 in PB. (D) Flow cytometric analysis of T effector cells (RA-RO+CD62L-CCR7-) in PB. (E-F) Analysis of Th1 (IFN-γ), Th2 (IL-4), Th17 (IL-17) secreting cell populations using either flow cytometry (E) or Luminex (F). (G) Flow cytometric analysis of NK cells in patient PB (CD3-CD56+). (H) Plasma analysis of IL-15 in patient PB.
Figure 4. Analysis of GVHD in patient…
Figure 4. Analysis of GVHD in patient plasma
Plasma analysis of inflammatory cytokines IL-2. (A) IL-6 (B) and IL-8 (C) in patient PB. Analysis of GVHD biomarkers ST-2 (D), osteopontin (E) and follistatin (F) in patient PB. (G-H) ELISA analysis of REG3a and Elafin in plasma of patient PB.

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Source: PubMed

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