Differential Effects of Calcineurin and Mammalian Target of Rapamycin Inhibitors on Alloreactive Th1, Th17, and Regulatory T Cells

Lorenzo Gallon, Opas Traitanon, Yuming Yu, Bo Shi, Joseph R. Leventhal, Joshua Miller, Valeria Mas, Xu L, James M. Mathew

Research output: Contribution to journalArticle

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Abstract

Previously, we had reported the role of tacrolimus (TAC) versus sirolimus (SRL) on the generation of regulatory T cells (Tregs) in primary MLR assays with SRL, demonstrating a uniquely supportive effect. However, the mechanisms associated with their actions on alloreactive human T cells are not fully understood. Therefore, we tested whether TAC and SRL differentially affect already alloactivated human CD4+ T-cell subsets. Methods Alloreactive CD4+CD45RA-/CD45RO+ T cells generated in 9-day MLR were cocultured with anti-CD3 and autologous antigen presenting cells plus interleukin (IL)-2 in presence of TAC, SRL, or both, and the Tregs generated after another 5 to 6 days were phenotypically, molecularly, and functionally characterized. Results Tacrolimus significantly and SRL modestly inhibited interferon (IFN)-γ (Th1) and IL-17 (Th17)-producing cells. At clinical therapeutic concentrations, SRL, however, significantly increased forkhead/winged helix transcription factor P3 (FOXP3+) Tregs, whereas TAC inhibited this T-cell population dose dependently and significantly. When used in combination, TAC and SRL had additive effects on inhibition of IFN-γ-and IL-17-producing cells. This was in contrast to the ability of SRL to reverse TAC-mediated inhibition of FOXP3-expressing cells. Proinflammatory cytokines (IL-1β, IL-6, and tumor necrosis factor-α) added to cultures caused significant decrease in FOXP3+ Tregs that was again reversed by SRL. Sirolimus-derived Tregs were phenotypically normal, anergic to allostimulation, and suppressed proliferation of allogeneic effector T-cells. Conclusions Thus, although TAC inhibits all alloreactive T cells, SRL promotes the differentiation and expansion of donor-specific Tregs without secondary reprogramming to IFN-γ+FOXP3+ and IL-17+FOXP3+ Treg subsets. These results, although performed in an artificial in vitro model, add clinically applicable information on how these agents affect T-cell subpopulations.

Original languageEnglish (US)
Pages (from-to)1774-1784
Number of pages11
JournalTransplantation
Volume99
Issue number9
DOIs
StatePublished - Sep 1 2015

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Interleukin-17
Calcineurin
Regulatory T-Lymphocytes
Sirolimus
Tacrolimus
T-Lymphocytes
Interferons
Winged-Helix Transcription Factors
CD3 Antigens
Autoantigens
T-Lymphocyte Subsets
Antigen-Presenting Cells
Interleukin-1
Interleukin-2
Interleukin-6
Tumor Necrosis Factor-alpha
Cytokines

All Science Journal Classification (ASJC) codes

  • Transplantation

Cite this

Differential Effects of Calcineurin and Mammalian Target of Rapamycin Inhibitors on Alloreactive Th1, Th17, and Regulatory T Cells. / Gallon, Lorenzo; Traitanon, Opas; Yu, Yuming; Shi, Bo; Leventhal, Joseph R.; Miller, Joshua; Mas, Valeria; L, Xu; Mathew, James M.

In: Transplantation, Vol. 99, No. 9, 01.09.2015, p. 1774-1784.

Research output: Contribution to journalArticle

Gallon, Lorenzo ; Traitanon, Opas ; Yu, Yuming ; Shi, Bo ; Leventhal, Joseph R. ; Miller, Joshua ; Mas, Valeria ; L, Xu ; Mathew, James M. / Differential Effects of Calcineurin and Mammalian Target of Rapamycin Inhibitors on Alloreactive Th1, Th17, and Regulatory T Cells. In: Transplantation. 2015 ; Vol. 99, No. 9. pp. 1774-1784.
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AU - Gallon, Lorenzo

AU - Traitanon, Opas

AU - Yu, Yuming

AU - Shi, Bo

AU - Leventhal, Joseph R.

AU - Miller, Joshua

AU - Mas, Valeria

AU - L, Xu

AU - Mathew, James M.

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N2 - Previously, we had reported the role of tacrolimus (TAC) versus sirolimus (SRL) on the generation of regulatory T cells (Tregs) in primary MLR assays with SRL, demonstrating a uniquely supportive effect. However, the mechanisms associated with their actions on alloreactive human T cells are not fully understood. Therefore, we tested whether TAC and SRL differentially affect already alloactivated human CD4+ T-cell subsets. Methods Alloreactive CD4+CD45RA-/CD45RO+ T cells generated in 9-day MLR were cocultured with anti-CD3 and autologous antigen presenting cells plus interleukin (IL)-2 in presence of TAC, SRL, or both, and the Tregs generated after another 5 to 6 days were phenotypically, molecularly, and functionally characterized. Results Tacrolimus significantly and SRL modestly inhibited interferon (IFN)-γ (Th1) and IL-17 (Th17)-producing cells. At clinical therapeutic concentrations, SRL, however, significantly increased forkhead/winged helix transcription factor P3 (FOXP3+) Tregs, whereas TAC inhibited this T-cell population dose dependently and significantly. When used in combination, TAC and SRL had additive effects on inhibition of IFN-γ-and IL-17-producing cells. This was in contrast to the ability of SRL to reverse TAC-mediated inhibition of FOXP3-expressing cells. Proinflammatory cytokines (IL-1β, IL-6, and tumor necrosis factor-α) added to cultures caused significant decrease in FOXP3+ Tregs that was again reversed by SRL. Sirolimus-derived Tregs were phenotypically normal, anergic to allostimulation, and suppressed proliferation of allogeneic effector T-cells. Conclusions Thus, although TAC inhibits all alloreactive T cells, SRL promotes the differentiation and expansion of donor-specific Tregs without secondary reprogramming to IFN-γ+FOXP3+ and IL-17+FOXP3+ Treg subsets. These results, although performed in an artificial in vitro model, add clinically applicable information on how these agents affect T-cell subpopulations.

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