Abstract
BackgroundThe aim of this study was to investigate the molecular mechanisms involved in the production of Th1 cytokines, namely IL-12 and IL-27, when the intra-macrophage redox state was altered by different chemical entities such as GSH-C4, which is reduced glutathione carrying an aliphatic chain, or I-152, a pro-drug of N-acetyl-cysteine (NAC) and beta-mercaptoethylamine. We had already demonstrated that GSH-C4 and I-152 could shift the immune response towards Th1 in Ovalbumin-immunized mice as well as enhance Th1 response in HIV-1 Tat-immunized mice.Methodology/Principal FindingsBy a new high performance liquid chromatography method, we found that 20 mM GSH-C4 provided a number of thiol species in the form of GSH, while 20 mM I-152 decreased GSH and increased the thiols in the form of NAC and I-152. Under these experimental conditions, GSH-C4 and I-152 enhanced and suppressed respectively the mRNA expression levels of IL-12 p40 induced by LPS/IFN-γ as assessed by Real-Time PCR. The protein production of IL-12 p40 was increased by GSH-C4 and decreased by I-152 as determined by Enzyme-linked immunosorbent assay. Western immunoblot and electrophoretic mobility shift assays revealed that Nuclear Factor -kB (NF-kB) activation was inhibited by I-152 and prolonged by GSH-C4. Twenty mM I-152 stimulated IL-27 p28 gene expression and sustained Signal Transducer and Activator of Transcription (STAT)-mediated interferon regulator factor 1 (IRF-1) de novo synthesis. By contrast, 20 mM GSH-C4 did not exert any effect on IL-27 p28 gene expression.Conclusions and Significancean increase in the intra-macrophage redox state by GSH-C4 and I-152 enhances Th1 cytokine production although the chemical structure and the intra-cellular metabolism influence differently signalling pathways involved in IL-27 or IL-12 production. GSH-C4 and I-152 may be used as Th1 immunomodulators in some pathologies and in ageing where GSH depletion may contribute to the Th1/Th2 imbalance, and in new immunization strategies.
Highlights
CD4+T helper cells differentiate into specialized types of effector cells: type 1 (Th1), type 2 (Th2) or the recently identified IL-17 producing (Th17) helper T cells, which promote different aspects of the immune response
Conclusions and Significance: an increase in the intra-macrophage redox state by GSH-C4 and I-152 enhances Th1 cytokine production the chemical structure and the intra-cellular metabolism influence differently signalling pathways involved in IL-27 or IL-12 production
GSH-C4 and I-152 may be used as Th1 immunomodulators in some pathologies and in ageing where GSH depletion may contribute to the Th1/Th2 imbalance, and in new immunization strategies
Summary
CD4+T helper cells differentiate into specialized types of effector cells: type 1 (Th1), type 2 (Th2) or the recently identified IL-17 producing (Th17) helper T cells, which promote different aspects of the immune response. In human monocyte-derived DCs (MD-DCs), the molecules able to increase and decrease intracellular GSH content could augment and reduce lipopolysaccharide (LPS)-induced IL-27 respectively [8] Expression of both IL-12 and IL-27 seem to be profoundly influenced by the redox state of APC. IL-27 activates STAT-1 and T-bet, two transcription factors that are important for Th1 polarization, and leads to up-regulation of the IL-12Rb2 chain In this way, IL-27 sensitizes the cells to Th1 enhancing cytokine IL-12. The aim of this study was to investigate the molecular mechanisms involved in the production of Th1 cytokines, namely IL-12 and IL-27, when the intra-macrophage redox state was altered by different chemical entities such as GSH-C4, which is reduced glutathione carrying an aliphatic chain, or I-152, a pro-drug of N-acetyl-cysteine (NAC) and betamercaptoethylamine. We had already demonstrated that GSH-C4 and I-152 could shift the immune response towards Th1 in Ovalbumin-immunized mice as well as enhance Th1 response in HIV-1 Tat-immunized mice
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