Effects of histone deacetylase inhibitor SAHA on effector and FOXP3+regulatory T cells in rhesus macaques

Abstract

Suberoylanilide hydroxamic acid (SAHA) a histone deacetylase inhibitor (HDACi), is clinically approved for treatment of cutaneous T-cell lymphoma. Although the exact underlying mechanisms are unknown, HDACi arrests the cell cycle in rapidly proliferating tumor cells and promote their apoptosis. HDACi were also recently shown to enhance the production and suppressive functions of Foxp3+ regulatory T (Treg) cells in rodents, leading us to begin to investigate the actions of HDACi on rhesus monkey T cells for the sake of potential preclinical applications. In this study, we show that SAHA inhibits polyclonal activation and proliferation of rhesus T cells and that the antiproliferative effects are due to inhibition of T-effector (Teff) cells and enhancement of Treg cells. Cryopreserved rhesus macaque splenocytes were CFSE labeled, stimulated with anti-CD3/anti-CD28 and cultured for 5 days in the presence of varying concentrations of SAHA. Samples were then costained to evaluate CD4 and CD8 expression. Concentrations of SAHA (10 and 5 micromol/L) were toxic to splenocytes. Proliferation was inhibited by 57% in CD4 cells and 47% in CD8 cells when unseparated splenocytes were cultured with 3 micromol/L SAHA. Effector cells alone showed decreased inhibition to proliferation when cultured with 3 micromol/L and 1 micromol/L SAHA when compared to Teff plus Treg cells. Our data suggest that SAHA can be used as part of an immunosuppressive protocol to enhance graft survival by limiting Teff cell proliferation as well as increasing Treg cells, thereby promoting tolerance.

Fig 1

Effects of SAHA on rhesus T cells in vitro. (a) Dose-dependent inhibition of splenocyte proliferation. (b) SAHA inhibited proliferation of both CD4 and CD8 T cells, and effects were enhanced when Tregs were included in the assays. (c) Flow cytometric analysis showed that SAHA increased the numbers of Foxp3+ T cells in splenocyte cultures.