Superior immune reconstitution using Treg-expanded donor cells versus PTCy treatment in preclinical HSCT models
Dietlinde Wolf, … , Krishna V. Komanduri, Robert B. Levy
Dietlinde Wolf, … , Krishna V. Komanduri, Robert B. Levy
Published October 18, 2018
Citation Information: JCI Insight. 2018;3(20):e121717. https://doi.org/10.1172/jci.insight.121717.
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Research Article Immunology Transplantation

Superior immune reconstitution using Treg-expanded donor cells versus PTCy treatment in preclinical HSCT models

Abstract

Posttransplant cyclophosphamide (PTCy) has been found to be effective in ameliorating acute graft-versus-host disease (GVHD) in patients following allogeneic hematopoietic stem cell transplantation (aHSCT). Adoptive transfer of high numbers of donor Tregs in experimental aHSCT has shown promise as a therapeutic modality for GVHD regulation. We recently described a strategy for in vivo Treg expansion targeting two receptors: TNFRSF25 and CD25. To date, there have been no direct comparisons between the use of PTCy and Tregs regarding outcome and immune reconstitution within identical groups of transplanted mice. Here, we assessed these two strategies and found both decreased clinical GVHD and improved survival long term. However, recipients transplanted with Treg-expanded donor cells (TrED) exhibited less weight loss early after HSCT. Additionally, TrED recipients demonstrated less thymic damage, significantly more recent thymic emigrants, and more rapid lymphoid engraftment. Three months after HSCT, PTCy-treated and TrED recipients showed tolerance to F1 skin allografts and comparable immune function. Overall, TrED was found superior to PTCy with regard to weight loss early after transplant and initial lymphoid engraftment. Based on these findings, we speculate that morbidity and mortality after transplant could be diminished following TrED transplant into aHSCT recipients, and, therefore, that TrED could provide a promising clinical strategy for GVHD prophylaxis.

Authors

Dietlinde Wolf, Cameron S. Bader, Henry Barreras, Sabrina Copsel, Brent J. Pfeiffer, Casey O. Lightbourn, Norman H. Altman, Krishna V. Komanduri, Robert B. Levy

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Figure 2

Comparison of TrED and PTCy treatment in a minor MHC-mismatch model shows comparable results to the major MHC-mismatch model.

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Comparison of TrED and PTCy treatment in a minor MHC-mismatch model show...
(A–C) A HSCT utilizing a B10.D2 BALB/c donor/recipient mouse model across a MHC-matched, minor histocompatibility antigen mismatch was performed using 8 × 106 non-TCD BM cells + 25 × 106 spleen cells from either untreated (GVHD and PTCy group) or TL1A-Ig/IL-2–expanded (TrED group) B10.D2 donor mice (BM: n = 8; GVHD and PTCy: n = 12; TrED: n = 17). Percentage of initial weight (day 10–24, P < 0.0001) (A), clinical score (day 10–24, P < 0.0001) (B) and survival (**P < 0.01) (C) are presented. Survival was analyzed by log-rank test. Statistical analysis for weights and clinical score was also performed to assess the overall model using JMP 13 Pro. P < 0.05 for all the groups. (D) Representative H&E staining from untreated recipient skin on day 31 after HSCT exhibited substantial fibrosis and thickening accompanied by moderate infiltration and inflammation with patchy necrosis. However, skin from PTC-treated recipients showed mild fibrosis with mild inflammation and some infiltrate and hyperplasia. In contrast, no fibrosis or thickening was observed with only mild inflammation in TrED recipients. Original magnification, ×200. Pathology scores are shown on the right (n = 3–4). Data are shown as mean ± SEM; ANOVA with Bonferroni correction was applied for multiple comparisons. *P < 0.05. Data are pooled from 2 independent experiments. Scale bars: 100 μm. (E) Representative H&E staining (chosen from 2 independent experiments) from recipient lung on day 200 after HSCT exhibited multifocal areas of moderate chronic, active inflammation, and fibrosis. The inflammation was characterized mainly by lymphocytes and macrophages with smaller numbers of neutrophils. Many alveoli also contained large macrophages and foamy cytoplasm in the PTCy compared with the TrED group, which was within normal limits. Original magnification, ×200. Pathology scores are shown on the right (n = 6–8). Data are shown as mean ± SEM; ANOVA with Bonferroni correction was applied for multiple comparisons. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001. Scale bars: 100 μm.