RESUMEN
The butyric acid derivative, 2-(4-morpholynl) ethyl butyrate hydrochloride (MEB), has been reported to induce antigen-specific T cell unresponsiveness and to block T cell-mediated graft-versus-host disease. As a potential therapeutic agent, it was important to determine the effects of MEB on other cells that contribute to immunopathology. Accordingly, we tested the effects of MEB on macrophage functions. MEB did not affect macrophage viability, phagocytic activity, or the activation-induced up-regulation of molecules associated with antigen presentation: MHC-II, CD86, CD40, or ICAM-1. However, MEB potently inhibited activation-induced production of inflammatory mediators, including tumor necrosis factor-alpha (TNF-alpha), IL-6, chemokine CCL2 and nitric oxide (NO). MEB inhibited the induction of NO synthase (NOS2), which is necessary for inducible NO, and inhibited nuclear translocation of NFkappaB, suggesting that MEB interferes with the signaling pathway involved in NOS2 induction. Thus, while inducing specific T cell unresponsiveness, MEB also exerts anti-inflammatory activity by acting on macrophages to suppress production of cytokines and NO.
Asunto(s)
Antígenos/inmunología , Butiratos/farmacología , Antagonistas de los Receptores Histamínicos/farmacología , Mediadores de Inflamación/metabolismo , Macrófagos/metabolismo , Morfolinas/farmacología , Linfocitos T/efectos de los fármacos , Animales , Antígenos CD/biosíntesis , Antígeno B7-2 , Western Blotting , Antígenos CD40/biosíntesis , Células Cultivadas , Citotoxicidad Inmunológica/efectos de los fármacos , Femenino , Citometría de Flujo , Genes MHC Clase II/efectos de los fármacos , Molécula 1 de Adhesión Intercelular/biosíntesis , Activación de Linfocitos/inmunología , Activación de Linfocitos/fisiología , Macrófagos/efectos de los fármacos , Glicoproteínas de Membrana/biosíntesis , Ratones , Ratones Endogámicos C57BL , FN-kappa B/efectos de los fármacos , Óxido Nítrico/metabolismo , Fagocitosis/efectos de los fármacosRESUMEN
MEB [n-butyrate 2-(4-morpholinyl) ethyl butyrate hydrochloride], a histone deacetylase inhibitor and G1 blocker, has been shown to induce unresponsiveness in antigen-activated Th1 cells. MEB was tested for here for its ability to inactivate naive alloantigen-specific T cells from DBA/2 and C57BL/10 mice. Since T cells from these two strains of mice have been shown to differ in their cell cycle regulation, it we hoped that this comparison would provide information concerning the role of cycle regulatory proteins in mediating MEB-induced T cell unresponsiveness. MEB inhibited proliferation in a one-way mixed lymphocyte reaction (MLR) in which spleen cells from DBA/2 mice (H-2d) or C57BL/10 mice (H-2b) were stimulated with spleen cells from C57BL/10 or DBA/2 mice, respectively. C57BL/10 responder T cells isolated from the MEB-treated primary MLR remained unresponsive to alloantigen following restimulation in a secondary MLR that did not contain MEB. T cells from DBA/2 mice were less sensitive to MEB-induced unresponsiveness and required a longer exposure or pretreatment with IL-2 to become tolerant. In all cases responsiveness to MEB-induced tolerance in the alloantigen-stimulated T cells corresponded with the levels of the cyclin-dependent kinase inhibitor p21cip1. Additional experiments showed that T cells from p21cip1-deficient mice, unlike T cells from p21cip1 wild-type littermates, were resistant to MEB-induced tolerance. These results underscore the role of p21cip1 in mediating T cell tolerance induced by the histone deacetylase inhibitor MEB.
Asunto(s)
Butiratos/farmacología , Linfocitos T CD4-Positivos/efectos de los fármacos , Linfocitos T CD8-positivos/efectos de los fármacos , Inhibidor p21 de las Quinasas Dependientes de la Ciclina/metabolismo , Inhibidores de Histona Desacetilasas , Morfolinas/farmacología , Subgrupos de Linfocitos T/efectos de los fármacos , Animales , Linfocitos T CD4-Positivos/enzimología , Linfocitos T CD8-positivos/enzimología , Ciclo Celular/efectos de los fármacos , Proliferación Celular , Células Cultivadas , Inhibidor p21 de las Quinasas Dependientes de la Ciclina/genética , Relación Dosis-Respuesta a Droga , Histona Desacetilasas/metabolismo , Histonas/metabolismo , Humanos , Tolerancia Inmunológica/efectos de los fármacos , Células Jurkat , Prueba de Cultivo Mixto de Linfocitos , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Endogámicos DBA , Ratones Noqueados , Especificidad de la Especie , Bazo/citología , Bazo/efectos de los fármacos , Bazo/enzimología , Subgrupos de Linfocitos T/enzimología , Factores de TiempoRESUMEN
Graft-versus-host-disease (GVHD) is a common and potentially fatal complication following bone marrow transplantation. This study was initiated to test whether MEB [n-butyrate 2-(4-morpholinyl) ethyl butyrate hydrochloride], a derivative of the G1 blocker butyric acid, could specifically inactivate the alloantigen-specific T cells that mediate GVHD. MEB was shown to inhibit proliferation in a one-way mixed lymphocyte reaction (MLR) in which spleen cells from C57BL/6 mice (H-2b) were stimulated with spleen cells from DBA/2 mice (H-2d). The addition of MEB to the MLR prevented the expansion of alloantigen-stimulated CD8+ and CD4+ T cells in association with decreased IL-2 production. In addition, MEB inhibited the CTL activity of CD8+ T cells from the MLR. Most importantly, T cells from the MEB-treated MLR, unlike T cells from an untreated MLR, were unable to induce the splenomegaly and increased serum TNF-alpha levels characteristic of acute GVHD when injected into B6D2F1 mice. The splenomegaly found in the B6D2F1 mice injected with T cells from an untreated MLR encompassed the expansion and activation of CD8+ T cells, CD4+ T cells, B cells and macrophages. In contrast, the spleens of mice injected with T cells from MEB-treated MLR looked essentially identical to those of control B6D2F1 mice in terms of the numbers and activation state of the spleen cell populations. Similarly, the increase in IFN-gamma and TNF-alpha production by CD4+ and CD8+ T cells from the spleens of mice undergoing acute GVHD was not observed if the mice were injected with T cells from an MEB-treated MLR instead of an untreated MLR. The use of MEB to inactivate host-specific T cells ex vivo underlines the potential clinical importance of this compound in the prevention and treatment of unwanted immune responses such as GVHD.