RESUMO
Ligation of retinoic acid receptor alpha (RARα) by RA promotes varied transcriptional programs associated with immune activation and tolerance, but genetic deletion approaches suggest the impact of RARα on TCR signaling. Here, we examined whether RARα would exert roles beyond transcriptional regulation. Specific deletion of the nuclear isoform of RARα revealed an RARα isoform in the cytoplasm of T cells. Extranuclear RARα was rapidly phosphorylated upon TCR stimulation and recruited to the TCR signalosome. RA interfered with extranuclear RARα signaling, causing suboptimal TCR activation while enhancing FOXP3+ regulatory T cell conversion. TCR activation induced the expression of CRABP2, which translocates RA to the nucleus. Deletion of Crabp2 led to increased RA in the cytoplasm and interfered with signalosome-RARα, resulting in impaired anti-pathogen immunity and suppressed autoimmune disease. Our findings underscore the significance of subcellular RA/RARα signaling in T cells and identify extranuclear RARα as a component of the TCR signalosome and a determinant of immune responses.
Assuntos
Doenças Autoimunes , Ativação Linfocitária , Humanos , Receptor alfa de Ácido Retinoico/genética , Membrana Celular , Receptores de Antígenos de Linfócitos TRESUMO
Vitamin A is a multifunctional vitamin implicated in a wide range of biological processes. Its control over the immune system and functions are perhaps the most pleiotropic not only for development but also for the functional fate of almost every cell involved in protective or regulatory adaptive or innate immunity. This is especially key at the intestinal border, where dietary vitamin A is first absorbed. Most effects of vitamin A are exerted by its metabolite, retinoic acid (RA), which through ligation of nuclear receptors controls transcriptional expression of RA target genes. In addition to this canonical function, RA and RA receptors (RARs), either as ligand-receptor or separately, play extranuclear, nongenomic roles that greatly expand the multiple mechanisms employed for their numerous and paradoxical functions that ultimately link environmental sensing with immune cell fate. This review discusses RA and RARs and their complex roles in innate and adaptive immunity.
Assuntos
Sistema Imunitário , Mucosa Intestinal/fisiologia , Receptores do Ácido Retinoico/imunologia , Tretinoína/metabolismo , Vitamina A/imunologia , Imunidade Adaptativa , Animais , Humanos , Imunidade Inata , Imunomodulação , Receptores do Ácido Retinoico/metabolismo , Tretinoína/imunologiaRESUMO
The generation of CD8(+) T cells by vaccination represents an important goal for protective immunity to infectious pathogens. It is thus of utmost importance to understand the mechanisms involved in the generation of optimal CD8(+) T-cell responses. The forkhead box O (FoxO) family of transcription factors has a crucial role in cellular responses to environmental change. Among them, FoxO3 is critically involved in the regulation of cellular proliferation, apoptosis, metabolism and stress resistance to withdrawal of nutrients or cytokine growth factors. Since the role of FoxO3 has been poorly studied in the immune system, here we have evaluated its involvement in the CD8(+) T-cell response. We observe that CD8(+) T cells deficient for FoxO3 undergo a significantly greater primary expansion than their wild-type (WT) counterparts in response to both infectious (vaccinia virus) or non-infectious (non-replicating cellular vaccine) immunogens, resulting in a larger cohort of cells following contraction. These survivors, however, do not undergo a greater secondary response than WT. Taken together, our data show that FoxO3 is a negative regulator of the CD8(+) T-cell response, specifically during the primary expansion.
Assuntos
Linfócitos T CD8-Positivos/citologia , Linfócitos T CD8-Positivos/imunologia , Fatores de Transcrição Forkhead/metabolismo , Memória Imunológica , Animais , Diferenciação Celular/imunologia , Proliferação de Células , Citocinas/biossíntese , Proteína Forkhead Box O3 , Contagem de Linfócitos , Camundongos Endogâmicos C57BLRESUMO
Coreceptor CD4 and CD8αß double-negative (DN) TCRαß(+) intraepithelial T cells, although numerous, have been greatly overlooked and their contribution to the immune response is not known. Here we used T cell receptor (TCR) sequencing of single cells combined with retrogenic expression of TCRs to study the fate and the major histocompatibility complex (MHC) restriction of DN TCRαß(+) intraepithelial T cells. The data show that commitment of thymic precursors to the DN TCRαß(+) lineage is imprinted by their TCR specificity. Moreover, the TCRs they express display a diverse and unusual pattern of MHC restriction that is nonoverlapping with that of CD4(+) or CD8αß(+) T cells, indicating that they sense antigens that are not recognized by the conventional T cell subsets. The new insights indicate that DN TCRαß(+) T cells form a third lineage of TCRαß T lymphocytes expressing a variable TCR repertoire, which serve nonredundant immune functions.
Assuntos
Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD8-Positivos/imunologia , Linhagem da Célula/imunologia , Receptores de Antígenos de Linfócitos T alfa-beta/genética , Receptores de Antígenos de Linfócitos T alfa-beta/imunologia , Animais , Diferenciação Celular/imunologia , Antígenos de Histocompatibilidade Classe I/imunologia , Antígenos de Histocompatibilidade Classe II/imunologia , Vigilância Imunológica/imunologia , Ativação Linfocitária/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Camundongos KnockoutRESUMO
The herpes virus entry mediator (HVEM), a member of the tumour-necrosis factor receptor family, has diverse functions, augmenting or inhibiting the immune response. HVEM was recently reported as a colitis risk locus in patients, and in a mouse model of colitis we demonstrated an anti-inflammatory role for HVEM, but its mechanism of action in the mucosal immune system was unknown. Here we report an important role for epithelial HVEM in innate mucosal defence against pathogenic bacteria. HVEM enhances immune responses by NF-κB-inducing kinase-dependent Stat3 activation, which promotes the epithelial expression of genes important for immunity. During intestinal Citrobacter rodentium infection, a mouse model for enteropathogenic Escherichia coli infection, Hvem−/− mice showed decreased Stat3 activation, impaired responses in the colon, higher bacterial burdens and increased mortality. We identified the immunoglobulin superfamily molecule CD160 (refs 7 and 8), expressed predominantly by innate-like intraepithelial lymphocytes, as the ligand engaging epithelial HVEM for host protection. Likewise, in pulmonary Streptococcus pneumoniae infection, HVEM is also required for host defence. Our results pinpoint HVEM as an important orchestrator of mucosal immunity, integrating signals from innate lymphocytes to induce optimal epithelial Stat3 activation, which indicates that targeting HVEM with agonists could improve host defence.
Assuntos
Citrobacter rodentium/imunologia , Citrobacter rodentium/patogenicidade , Imunidade nas Mucosas/imunologia , Mucosa/imunologia , Mucosa/microbiologia , Membro 14 de Receptores do Fator de Necrose Tumoral/metabolismo , Transdução de Sinais , Animais , Antígenos CD/imunologia , Antígenos CD/metabolismo , Carga Bacteriana , Linhagem Celular , Modelos Animais de Doenças , Infecções por Enterobacteriaceae/imunologia , Infecções por Enterobacteriaceae/microbiologia , Escherichia coli Enteropatogênica , Células Epiteliais/imunologia , Células Epiteliais/metabolismo , Infecções por Escherichia coli , Proteínas Ligadas por GPI/imunologia , Proteínas Ligadas por GPI/metabolismo , Intestinos/imunologia , Intestinos/microbiologia , Ligantes , Pulmão/imunologia , Pulmão/microbiologia , Linfócitos/imunologia , Linfócitos/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Mucosa/metabolismo , Infecções Pneumocócicas/imunologia , Infecções Pneumocócicas/microbiologia , Proteínas Serina-Treonina Quinases/metabolismo , Receptores Imunológicos/imunologia , Receptores Imunológicos/metabolismo , Membro 14 de Receptores do Fator de Necrose Tumoral/deficiência , Membro 14 de Receptores do Fator de Necrose Tumoral/genética , Membro 14 de Receptores do Fator de Necrose Tumoral/imunologia , Fator de Transcrição STAT3/metabolismo , Streptococcus pneumoniae/imunologia , Taxa de Sobrevida , Quinase Induzida por NF-kappaBRESUMO
GCs are widely prescribed to treat inflammatory disorders and autoimmune and allergic diseases. Their anti-inflammatory and immunosuppressive effects may be related, in part, to their ability to control the maturation and functions of DCs. Here, we report that GCs inhibit the maturation of human CD34-DCs induced by the TLR7 agonist imiquimod and the TLR8 agonist 3M-002. GCs down-regulate the expression of CD86, CD40, CD83, CCR7, and HLA-DR on DCs and inhibit IL-6 and IL-12p40 production by DCs following TLR7 and TLR8 stimulation. This inhibitory effect is abolished by RU486, suggesting a role for GR transcriptional activity. Our results also show that GCs do not affect TLR-mediated DNA-binding activity of NF-κBp65. We observe that GCs control the activation of JNK induced by TLR agonists, without affecting its upstream MKK4. However, p38MAPK activation is not affected by GCs. Concomitantly to JNK inhibition, we observe the induction of the DUSP MKP-1 but not of other DUSPs by GCs. However, although silencing of MKP-1 in DCs reverses GC-mediated JNK inhibition, no significant effect on GC-induced inhibition of DC maturation was evidenced. Our results show that GCs alter DC maturation in response to TLR7 or TLR8 through a mechanism involving GR transcriptional activity.
Assuntos
Diferenciação Celular/imunologia , Células Dendríticas/efeitos dos fármacos , Células Dendríticas/imunologia , Dexametasona/farmacologia , Glucocorticoides/farmacologia , Receptor 7 Toll-Like/fisiologia , Receptor 8 Toll-Like/fisiologia , Diferenciação Celular/efeitos dos fármacos , Células Cultivadas , Células Dendríticas/citologia , Sangue Fetal/citologia , Sangue Fetal/imunologia , Inibidores do Crescimento/farmacologia , Humanos , Receptor 7 Toll-Like/antagonistas & inibidores , Receptor 8 Toll-Like/antagonistas & inibidoresRESUMO
The presence of immune memory at pathogen-entry sites is a prerequisite for protection. Nevertheless, the mechanisms that warrant immunity at peripheral interfaces are not understood. Here we show that the nonclassical major histocompatibility complex (MHC) class I molecule thymus leukemia antigen (TL), induced on dendritic cells interacting with CD8αα on activated CD8αß(+) T cells, mediated affinity-based selection of memory precursor cells. Furthermore, constitutive expression of TL on epithelial cells led to continued selection of mature CD8αß(+) memory T cells. The memory process driven by TL and CD8αα was essential for the generation of CD8αß(+) memory T cells in the intestine and the accumulation of highly antigen-sensitive CD8αß(+) memory T cells that form the first line of defense at the largest entry port for pathogens.
Assuntos
Células Dendríticas/metabolismo , Listeriose/imunologia , Glicoproteínas de Membrana/metabolismo , Células Precursoras de Linfócitos T/metabolismo , Linfócitos T/metabolismo , Animais , Antígenos/imunologia , Antígenos/metabolismo , Antígenos CD8/metabolismo , Diferenciação Celular , Seleção Clonal Mediada por Antígeno , Células Dendríticas/imunologia , Células Dendríticas/patologia , Imunidade nas Mucosas/genética , Memória Imunológica/genética , Ativação Linfocitária/genética , Glicoproteínas de Membrana/genética , Glicoproteínas de Membrana/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Células Precursoras de Linfócitos T/imunologia , Células Precursoras de Linfócitos T/patologia , Linfócitos T/imunologia , Linfócitos T/patologia , Transgenes/genéticaRESUMO
Allergic contact dermatitis, caused by metallic ions, is a T cell-mediated inflammatory skin disease. IL-12 is a 70-kDa heterodimeric protein composed of IL-12p40 and IL-12p35, playing a major role in the generation of allergen-specific T cell responses. Dendritic cells (DCs) are APCs involved in the induction of primary immune responses, as they possess the ability to stimulate naive T cells. In this study, we address the question whether the sensitizer nickel sulfate (NiSO(4)) itself or in synergy with other signals can induce the secretion of IL-12p70 in human monocyte-derived DCs (Mo-DCs). We found that IL-12p40 was produced by Mo-DC in response to NiSO(4) stimulation. Addition of IFN-gamma concomitantly to NiSO(4) leads to IL-12p70 synthesis. NiSO(4) treatment leads to the activation of MAPK, NF-kappaB pathways, and IFN regulatory factor 1 (IRF-1). We investigated the role of these signaling pathways in IL-12 production using known pharmacological inhibitors of MAPK and NF-kappaB pathways and RNA interference-mediated silencing of IRF-1. Our results showed that p38 MAPK, NF-kappaB, and IRF-1 were involved in IL-12p40 production induced by NiSO(4). Moreover, IRF-1 silencing nearly totally abrogated IL-12p40 and IL-12p70 production provoked by NiSO(4) and IFN-gamma. In response to NiSO(4), we observed that STAT-1 was phosphorylated on both serine and tyrosine residues and participated to NiSO(4)-induced IRF-1 activation. N-acetylcysteine abolished STAT-1 phosphorylation, suggesting that STAT-1 activation may be dependent on NiSO(4)-induced alteration of the redox status of the cell. These results indicate that p38 MAPK, NF-kappaB, and IRF-1 are activated by NiSO(4) in Mo-DC and cooperate for IL-12 production.
Assuntos
Células Dendríticas/imunologia , Células Dendríticas/metabolismo , Interleucina-1/biossíntese , Irritantes/toxicidade , Níquel/toxicidade , Células Cultivadas , Células Dendríticas/enzimologia , Humanos , Fator Regulador 1 de Interferon/antagonistas & inibidores , Fator Regulador 1 de Interferon/metabolismo , Fator Regulador 1 de Interferon/fisiologia , Interferon gama/fisiologia , Interleucina-12/biossíntese , Interleucina-12/metabolismo , Subunidade p35 da Interleucina-12/biossíntese , Subunidade p35 da Interleucina-12/genética , Subunidade p40 da Interleucina-12/biossíntese , Proteínas Quinases JNK Ativadas por Mitógeno/fisiologia , Sistema de Sinalização das MAP Quinases/imunologia , Monócitos/enzimologia , Monócitos/imunologia , Monócitos/metabolismo , NF-kappa B/antagonistas & inibidores , NF-kappa B/metabolismo , NF-kappa B/fisiologia , RNA Mensageiro/biossíntese , Fator de Transcrição STAT1/fisiologia , Proteínas Quinases p38 Ativadas por Mitógeno/antagonistas & inibidores , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo , Proteínas Quinases p38 Ativadas por Mitógeno/fisiologiaRESUMO
Dendritic cells (DCs) play an important role in bridging innate and adaptive immunity. These APCs have the ability to recognize specific molecular signatures of pathogens through TLRs. In particular, the intracellular TLR7 and TLR8, mediating the recognition of ssRNA by DCs, play a major role in the immune response during viral infection. Although differences have been identified between TLR7 and TLR8, in terms of cellular expression and functions, the signaling pathways that lead to DC maturation following TLR7 or TLR8 engagement are largely unknown. We compared the signaling pathways involved in human CD34-DC maturation induced by agonists selective for TLR7 (imiquimod) or TLR8 (3M002). TLR7 and TLR8 activation up-regulated CCR7, CD40, CD86, and CD83 expression and IL-6 and IL-12p40 production. However, only TLR8 activation led to IL-12p70 production and il-12p35 mRNA expression. We found that upon TLR7 and TLR8 activation, JNK and NF-kappaB positively regulated the expression of CCR7, CD86, CD83, and CD40 and the production of IL-6 and IL-12p40. However, although p38MAPK participated in the up-regulation of maturation markers in response to TLR7 activation, this kinase exerted an inhibitory effect on CD40 expression and IL-12 production in TLR8-stimulated DCs. We also showed that the Jak/STAT signaling pathway was involved in CD40 expression and cytokine production in TLR7-stimulated DCs but negatively regulated CD83 expression and cytokine secretion in DCs activated through TLR8. This study showed that TLR7 and TLR8 activate similar signaling pathways that play different roles in DC maturation, depending on which TLR is triggered.
Assuntos
Células Dendríticas/citologia , Transdução de Sinais/imunologia , Receptor 7 Toll-Like/agonistas , Receptor 8 Toll-Like/agonistas , Antígenos CD , Células Dendríticas/metabolismo , Humanos , Interleucinas , Proteínas Quinases , Receptores de Quimiocinas , Regulação para Cima/imunologiaRESUMO
Dendritic cells (DCs) play a major role in the regulation of immune responses to a variety of antigens (Ag) and haptens which participate in the process of DC maturation. Indeed, metallic haptens are able to induce DC maturation in vitro but the mechanism of this maturation is not well understood. We and others have already shown that NiSO(4) activates p38 mitogen-activated protein kinases (p38MAPK), c-jun N-terminal kinase (JNK), extracellular signal-regulated kinase (ERK) and the transcription factor NF-kappaB during the early events of DCs maturation. However, the effect of other metallic haptens on DC maturation is still poorly understood. In the present study, using dendritic cells derived from CD34(+) cord blood cells, we showed that both NiSO(4) and CoCl(2) induced the expression of CD86, CD83, HLA-DR and CD40 and the production of IL-6 in human DCs while K(2)Cr(2)O(7) induced only a slight upregulation of CD86. Interestingly, only NiSO(4) was able to induce the production of IL-12p40. NiSO(4) and CoCl(2) but not K(2)Cr(2)O(7) were able to activate the MAPK pathway and the transcription factor NF-kappaB. The role of MAPKs in metals-induced DC maturation was then evaluated using well-described pharmacological inhibitors. Our results suggest that p38MAPK activation regulates the expression of CD86 and CD83 induced by NiSO(4) while it only affects the expression of CD83 induced by CoCl(2). IL-6 production induced by NiSO(4) and CoCl(2) strongly depended on all MAPKs. IL-12p40 synthesis after NiSO(4) treatment was regulated by both p38MAPK and JNK pathways whereas ERK may play an inhibitory role. Our results show that both NiSO(4) and CoCl(2) activate similar signaling pathways that are playing different roles in DC maturation depending on the hapten used.