Calcineurin inhibition rescues alloantigen-specific central memory T cell subsets that promote chronic GVHD
Yewei Wang, … , Ping Zhang, Geoffrey R. Hill
Yewei Wang, … , Ping Zhang, Geoffrey R. Hill
Published June 3, 2024
Citation Information: J Clin Invest. 2024;134(11):e170125. https://doi.org/10.1172/JCI170125.
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Research Article Immunology

Calcineurin inhibition rescues alloantigen-specific central memory T cell subsets that promote chronic GVHD

Abstract

Calcineurin inhibitors (CNIs) constitute the backbone of modern acute graft-versus-host disease (aGVHD) prophylaxis regimens but have limited efficacy in the prevention and treatment of chronic GVHD (cGVHD). We investigated the effect of CNIs on immune tolerance after stem cell transplantation with discovery-based single-cell gene expression and T cell receptor (TCR) assays of clonal immunity in tandem with traditional protein-based approaches and preclinical modeling. While cyclosporin and tacrolimus suppressed the clonal expansion of CD8+ T cells during GVHD, alloreactive CD4+ T cell clusters were preferentially expanded. Moreover, CNIs mediated reversible dose-dependent suppression of T cell activation and all stages of donor T cell exhaustion. Critically, CNIs promoted the expansion of both polyclonal and TCR-specific alloreactive central memory CD4+ T cells (TCM) with high self-renewal capacity that mediated cGVHD following drug withdrawal. In contrast to posttransplant cyclophosphamide (PT-Cy), CSA was ineffective in eliminating IL-17A–secreting alloreactive T cell clones that play an important role in the pathogenesis of cGVHD. Collectively, we have shown that, although CNIs attenuate aGVHD, they paradoxically rescue alloantigen-specific TCM, especially within the CD4+ compartment in lymphoid and GVHD target tissues, thus predisposing patients to cGVHD. These data provide further evidence to caution against CNI-based immune suppression without concurrent approaches that eliminate alloreactive T cell clones.

Authors

Yewei Wang, Md Ashik Ullah, Olivia G. Waltner, Shruti S. Bhise, Kathleen S. Ensbey, Christine R. Schmidt, Samuel R.W. Legg, Tomoko Sekiguchi, Ethan L. Nelson, Rachel D. Kuns, Nicole S. Nemychenkov, Erden Atilla, Albert C. Yeh, Shuichiro Takahashi, Julie R. Boiko, Antiopi Varelias, Bruce R. Blazar, Motoko Koyama, Simone A. Minnie, Andrew D. Clouston, Scott N. Furlan, Ping Zhang, Geoffrey R. Hill

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Figure 2

Higher-dose CSA inhibits the acquisition of effector function but promotes survival and central memory differentiation.

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Higher-dose CSA inhibits the acquisition of effector function but promot...
(A–H) B6D2F1 recipients were transplanted with B6 BM (5 × 106) + T cells (2 × 106) and treated with saline or CSA (5 or 50 mg/kg/d) until day 13. Spleens were taken for immune phenotyping and donor T cells were gated with congenic markers. (A) Frequency of donor T cells in the spleens over time after BMT. (B and C) Ratio of CD4+ to CD8+ donor T cells on day 5 (B) and day 7 (C) (n = 10–11 per group from 2 experiments). (D) Representative flow cytometric plots and expression of caspase-3 and Ki-67 (relative to the saline group) in donor T cells on day 5 (n = 9–10 per group from 2 experiments). (E and F) Number of cytokine-producing donor T cells on day 5 (n = 9–10 per group from 2 experiments) (E) and day 17 (F) (n = 12–13 per group from 3 experiments). (G and H) Number of donor TN (CD44–CD62L+), TEM (CD44+CD62L–), and TCM (CD44+CD62L+) cells on day 5 (G) and day 17 with representative flow cytometric plots (H) (n = 9–13 per group from 2 experiments). (I and J) Experiments were set up as described in Supplemental Figure 1I whereby magnetic activated cell sorting–selected (MACS-selected) CD4+ and CD8+ T cells were combined from saline/CSA-treated mice and transferred to secondary recipients. Memory phenotype (I) and numbers of TCM subset (J) of the transferred T cells were determined 35 days after transfer (n = 8–10 per group from 2 experiments). (K) Donor CD4+ T cells were isolated from spleens on day 15 after BMT (saline or CSA treated), sort-purified to TN, TEM and TCM subsets, and individually transferred to secondary BMT recipients (1 × 105 per recipient) on day 0. Spleens were taken 49 days after adoptive transfer and transferred T cells were determined based on congenic markers (n = 2, 3, 6, 5, 6, and 4 per group respectively). (L) B6D2F1 recipients were transplanted with B6 BM (5 × 106) + T cells (2 × 106), treated with saline or CSA (50 mg/kg/d) up until day 13 and monitored for survival and clinical scores (n = 12–13 per group from 2 experiments). (A–K) Data are presented as median ± interquartile range and analyzed with the Mann-Whitney U test; (L) clinical score data are presented as mean ± SEM and survival analyzed by log rank. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001.