RESUMO
Dendritic cells (DCs) are critical in immune responses, linking innate and adaptive immunity. We found here that DC-specific deletion of the transcription factor STAT5 was not critical for development but was required for T helper type 2 (TH2), but not TH1, allergic responses in both the skin and lungs. Loss of STAT5 in DCs led to the inability to respond to thymic stromal lymphopoietin (TSLP). STAT5 was required for TSLP-dependent DC activation, including upregulation of the expression of costimulatory molecules and chemokine production. Furthermore, TH2 responses in mice with DC-specific loss of STAT5 resembled those seen in mice deficient in the receptor for TSLP. Our results show that the TSLP-STAT5 axis in DCs is a critical component for the promotion of type 2 immunity at barrier surfaces.
Assuntos
Células Dendríticas/imunologia , Células Dendríticas/metabolismo , Fator de Transcrição STAT5/metabolismo , Células Th2/imunologia , Animais , Diferenciação Celular , Citocinas/imunologia , Citocinas/metabolismo , Células Dendríticas/citologia , Dermatite de Contato/imunologia , Dermatite de Contato/metabolismo , Derme/imunologia , Derme/metabolismo , Feminino , Homeostase/imunologia , Janus Quinases/metabolismo , Pulmão/imunologia , Pulmão/metabolismo , Camundongos , Camundongos Knockout , Fator de Transcrição STAT5/genética , Transdução de Sinais , Células Th1/imunologia , Linfopoietina do Estroma do TimoRESUMO
Regulatory T (Treg) cells, driven by the Foxp3 transcription factor, are responsible for limiting autoimmunity and chronic inflammation. We showed that a well-characterized Foxp3(gfp) reporter mouse, which expresses an N-terminal GFP-Foxp3 fusion protein, is a hypomorph that causes profoundly accelerated autoimmune diabetes on a NOD background. Although natural Treg cell development and in vitro function are not markedly altered in Foxp3(gfp) NOD and C57BL/6 mice, Treg cell function in inflammatory environments was perturbed and TGF-ß-induced Treg cell development was reduced. Foxp3(gfp) was unable to interact with the histone acetyltransferase Tip60, the histone deacetylase HDAC7, and the Ikaros family zinc finger 4, Eos, which led to reduced Foxp3 acetylation and enhanced K48-linked polyubiquitylation. Collectively this results in an altered transcriptional landscape and reduced Foxp3-mediated gene repression, notably at the hallmark IL-2 promoter. Loss of controlled Foxp3-driven epigenetic modification leads to Treg cell insufficiency that enables autoimmunity in susceptible environments.
Assuntos
Fatores de Transcrição Forkhead/imunologia , Fatores de Transcrição Forkhead/metabolismo , Linfócitos T Reguladores/imunologia , Fatores de Transcrição/imunologia , Fatores de Transcrição/metabolismo , Acetilação , Animais , Autoimunidade/genética , Autoimunidade/imunologia , Proteínas de Transporte/genética , Proteínas de Transporte/imunologia , Proteínas de Transporte/metabolismo , Proteínas de Ligação a DNA , Diabetes Mellitus Tipo 1/genética , Diabetes Mellitus Tipo 1/imunologia , Diabetes Mellitus Tipo 1/metabolismo , Epigênese Genética , Fatores de Transcrição Forkhead/genética , Histona Acetiltransferases/genética , Histona Acetiltransferases/imunologia , Histona Acetiltransferases/metabolismo , Histona Desacetilases/genética , Histona Desacetilases/imunologia , Histona Desacetilases/metabolismo , Interleucina-2/genética , Interleucina-2/imunologia , Interleucina-2/metabolismo , Lisina Acetiltransferase 5 , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos NOD , Proteínas do Tecido Nervoso/genética , Proteínas do Tecido Nervoso/imunologia , Proteínas do Tecido Nervoso/metabolismo , Regiões Promotoras Genéticas , Linfócitos T Reguladores/metabolismo , Transativadores/genética , Transativadores/imunologia , Transativadores/metabolismo , Fatores de Transcrição/genética , Fator de Crescimento Transformador beta/genética , Fator de Crescimento Transformador beta/imunologia , Fator de Crescimento Transformador beta/metabolismoRESUMO
We tested the role of Stat5 in dendritic cell and alveolar macrophage (AM) homeostasis in the lung using CD11c-cre mediated deletion (Cre+5f/f). We show that Stat5 is required for CD103+ dendritic cell and AM development. We found that fetal monocyte maturation into AMs was impaired in Cre+5f/f mice, and we also confirmed impaired AM development of progenitor cells using mixed chimera experiments. In the absence of Stat5 signaling in AMs, mice developed alveolar proteinosis with altered lipid homeostasis. In addition, loss of Stat5 in CD11c+ cells was associated with exaggerated LPS-induced inflammatory responses and vascular leak. In Cre+5f/f mice, there was loss of immune-dampening effects on epithelial cells, a key source of CCL2 that serves to recruit monocytes and macrophages. These findings demonstrate the critical importance of Stat5 signaling in maintaining lung homeostasis, and underscore the importance of resident macrophages in moderating tissue damage and excess inflammation.
Assuntos
Antígenos CD/imunologia , Células Dendríticas/fisiologia , Cadeias alfa de Integrinas/imunologia , Lesão Pulmonar/imunologia , Macrófagos Alveolares/fisiologia , Fator de Transcrição STAT5/metabolismo , Animais , Antígenos CD/genética , Líquido da Lavagem Broncoalveolar/citologia , Líquido da Lavagem Broncoalveolar/imunologia , Células Dendríticas/imunologia , Células Epiteliais/imunologia , Inflamação/imunologia , Cadeias alfa de Integrinas/genética , Macrófagos Alveolares/imunologia , Camundongos , Monócitos/imunologia , Proteinose Alveolar Pulmonar/imunologiaRESUMO
Caspase-8 (casp8) is required for extrinsic apoptosis, and mice deficient in casp8 fail to develop and die in utero while ultimately failing to maintain the proliferation of T cells, B cells, and a host of other cell types. Paradoxically, these failures are not caused by a defect in apoptosis, but by a presumed proliferative function of this protease. Indeed, following mitogenic stimulation, T cells lacking casp8 or its adaptor protein FADD (Fas-associated death domain protein) develop a hyperautophagic morphology, and die a programmed necrosis-like death process termed necroptosis. Recent studies have demonstrated that receptor-interacting protein kinases (RIPKs) RIPK1 and RIPK3 together facilitate TNF-induced necroptosis, but the precise role of RIPKs in the demise of T cells lacking FADD or casp8 activity is unknown. Here we demonstrate that RIPK3 and FADD have opposing and complementary roles in promoting T-cell clonal expansion and homeostasis. We show that the defective proliferation of T cells bearing an interfering form of FADD (FADDdd) is rescued by crossing with RIPK3(-/-) mice, although such rescue ultimately leads to lymphadenopathy. Enhanced recovery of these double-mutant T cells following stimulation demonstrates that FADD, casp8, and RIPK3 are all essential for clonal expansion, contraction, and antiviral responses. Finally, we demonstrate that caspase-mediated cleavage of RIPK1-containing necrosis inducing complexes (necrosomes) is sufficient to prevent necroptosis in the face of death receptor signaling. These studies highlight the "two-faced" nature of casp8 activity, promoting clonal expansion in some situations and apoptotic demise in others.
Assuntos
Linfócitos T CD8-Positivos/enzimologia , Linfócitos T CD8-Positivos/virologia , Proteína de Domínio de Morte Associada a Fas/metabolismo , Homeostase/imunologia , Imunidade/imunologia , Vírus da Hepatite Murina/imunologia , Proteína Serina-Treonina Quinases de Interação com Receptores/metabolismo , Animais , Linfócitos T CD8-Positivos/citologia , Proliferação de Células , Sobrevivência Celular , Cruzamentos Genéticos , Feminino , Hepatite Viral Animal/imunologia , Hepatite Viral Animal/virologia , Masculino , Camundongos , Proteína Serina-Treonina Quinases de Interação com Receptores/deficiênciaRESUMO
Fas-associated death domain protein (FADD) and caspase-8 (casp8) are vital intermediaries in apoptotic signaling induced by tumor necrosis factor family ligands. Paradoxically, lymphocytes lacking FADD or casp8 fail to undergo normal clonal expansion following antigen receptor cross-linking and succumb to caspase-independent cell death upon activation. Here we show that T cells lacking FADD or casp8 activity are subject to hyperactive autophagic signaling and subvert a cellular survival mechanism into a potent death process. T cell autophagy, enhanced by mitogenic signaling, recruits casp8 through interaction with FADD:Atg5-Atg12 complexes. Inhibition of autophagic signaling with 3-methyladenine, dominant-negative Vps34, or Atg7 shRNA rescued T cells expressing a dominant-negative FADD protein. The necroptosis inhibitor Nec-1, which blocks receptor interacting protein kinase 1 (RIP kinase 1), also completely rescued T cells lacking FADD or casp8 activity. Thus, while autophagy is necessary for rapid T cell proliferation, our findings suggest that FADD and casp8 form a feedback loop to limit autophagy and prevent this salvage pathway from inducing RIPK1-dependent necroptotic cell death. Thus, linkage of FADD and casp8 to autophagic signaling intermediates is essential for rapid T cell clonal expansion and may normally serve to promote caspase-dependent apoptosis under hyperautophagic conditions, thereby averting necrosis and inflammation in vivo.
Assuntos
Autofagia , Caspase 8/fisiologia , Proliferação de Células , Proteína de Domínio de Morte Associada a Fas/fisiologia , Linfócitos T/citologia , Animais , Apoptose , Caspase 8/genética , Proteína de Domínio de Morte Associada a Fas/genética , Retroalimentação Fisiológica , Ativação Linfocitária , Camundongos , Camundongos Transgênicos , Transdução de Sinais/imunologiaRESUMO
Patients exhibit distinct responses to immunotherapies that are thought to be linked to their tumor immune environment. However, wide variations in outcomes are also observed in patients with matched baseline tumor environments, indicating that the biological response to treatment is not currently predictable using a snapshot analysis. To investigate the relationship between the immune environment of tumors and the biological response to immunotherapies, we characterized four murine head and neck squamous cell carcinoma (HNSCC) models on two genetic backgrounds. Using tumor explants from those models, we identified correlations between the composition of infiltrating immune cells and baseline cytokine profiles prior to treatment. Following treatment with PD-1 blockade, CTLA-4 blockade, or OX40 stimulation, we observed inter-individual variability in the response to therapy between genetically identical animals bearing the same tumor. These distinct biological responses to treatment were not linked to the initial tumor immune environment, meaning that outcome would not be predictable from a baseline analysis of the tumor infiltrates. We similarly performed the explant assay on patient HNSCC tumors and found significant variability between the baseline environment of the tumors and their response to therapy. We propose that tumor explants provide a rapid biological assay to assess response to candidate immunotherapies that may allow matching therapies to individual patient tumors. Further development of explant approaches may allow screening and monitoring of treatment responses in HNSCC.
RESUMO
Ankle involvement by synovial chondromatosis is unusual. It is unknown whether a post-traumatic event to the ankle induces the formation and development of these lesions. Synovial osteochondromatosis associated with post-traumatic ankle events are rare but suggest trauma to the synovial tissues as being causative, although this has never been statistically confirmed owing to the lack of reports and frequency. We report a case of primary synovial osteochondromatosis involving the tibiotalar joint with painful symptoms after a history of ankle injury, including magnetic resonance imaging findings of this unusual condition.
Assuntos
Traumatismos do Tornozelo/complicações , Condromatose Sinovial/etiologia , Entorses e Distensões/complicações , Adulto , Articulação do Tornozelo/patologia , Articulação do Tornozelo/cirurgia , Condromatose Sinovial/diagnóstico , Condromatose Sinovial/cirurgia , Humanos , Corpos Livres Articulares/patologia , Imageamento por Ressonância Magnética , MasculinoRESUMO
Surface micron-scale and submicron scale features increase osteoblast differentiation and enhance responses of osteoblasts to 1,25-dihydroxyvitamin D(3) [1alpha,25(OH)(2)D(3)]. beta(1) integrin expression is increased in osteoblasts grown on Ti substrates with rough microarchitecture, and it is regulated by 1alpha,25(OH)(2)D(3) in a surface-dependent manner. To determine if beta(1) has a role in mediating osteoblast response, we silenced beta(1) expression in MG63 human osteoblast-like cells using small interfering RNA (siRNA). In addition, MG63 cells were treated with two different monoclonal antibodies to human beta(1) to block ligand binding. beta(1)-silenced MG63 cells grown on a tissue culture plastic had reduced alkaline phosphatase activity and levels of osteocalcin, transforming growth factor beta(1), prostaglandin E(2), and osteoprotegerin in comparison with control cells. Moreover, beta(1)-silencing inhibited the effects of surface roughness on these parameters and partially inhibited effects of 1alpha,25(OH)(2)D(3). Anti beta(1) antibodies decreased alkaline phosphatase but increase osteocalcin; effects of 1alpha,25(OH)(2)D(3) on cell number and alkaline phosphatase were reduced and effects on osteocalcin were increased. These findings indicate that beta(1) plays a major and complex role in osteoblastic differentiation modulated by either surface microarchitecture or 1alpha,25(OH)(2)D(3). The results also show that beta(1) mediates, in part, the synergistic effects of surface roughness and 1alpha,25(OH)(2)D(3).
Assuntos
Calcitriol/metabolismo , Cadeias beta de Integrinas/metabolismo , Osteoblastos/metabolismo , Humanos , Cadeias beta de Integrinas/genética , Especificidade por SubstratoRESUMO
Surface microroughness increases osteoblast differentiation and enhances responses of osteoblasts to 1,25-dihydroxyvitamin D3 [1alpha,25(OH)2D3]. The observations that beta1 integrin expression is increased in osteoblasts grown on Ti substrates with rough microarchitecture, and that it is regulated by 1alpha,25(OH)2D3 in a surface-dependent manner, suggest that beta1 may play a role in mediating osteoblast response. To test this hypothesis, we silenced beta1 expression in MG63 human osteoblast-like cells using small interfering RNA (siRNA) and examined the responses of the beta1-silenced osteoblasts to surface microtopography and 1alpha,25(OH)2D3. To better understand the role of beta1, MG63 cells were also treated with two different monoclonal antibodies to human beta1 to block ligand binding. beta1-silenced MG63 cells grown on a tissue culture plastic had reduced alkaline phosphatase activity and levels of osteocalcin, transforming growth factor beta1, prostaglandin E2, and osteoprotegerin in comparison with control cells. Moreover, beta1-silencing inhibited the effects of surface roughness on these parameters and partially inhibited effects of 1alpha,25(OH)2D3. Anti beta1 antibody AIIB2 had no significant effect on cell number and osteocalcin, but decreased alkaline phosphatase; MAB2253Z caused dose-dependent decreases in cell number and alkaline phosphatase and an increase in osteocalcin. Effects of 1alpha,25(OH)2D3 on cell number and alkaline phosphatase were reduced and effects on osteocalcin were increased. These findings indicate that beta1 plays a major and complex role in osteoblastic differentiation modulated by either surface microarchitecture or 1alpha,25(OH)2D3. The results also show that beta1 mediates, in part, the synergistic effects of surface roughness and 1alpha,25(OH)2D3.
Assuntos
Conservadores da Densidade Óssea/metabolismo , Calcitriol/metabolismo , Inativação Gênica , Integrina beta1/metabolismo , Osteoblastos/fisiologia , Animais , Anticorpos/metabolismo , Técnicas de Cultura de Células , Linhagem Celular , Dinoprostona/genética , Dinoprostona/metabolismo , Humanos , Integrina beta1/genética , Camundongos , Osteoblastos/citologia , Osteoprotegerina/genética , Osteoprotegerina/metabolismo , RNA Interferente Pequeno/metabolismo , Propriedades de Superfície , Titânio/metabolismo , Fator de Crescimento Transformador beta1/genética , Fator de Crescimento Transformador beta1/metabolismoRESUMO
The design of ecotoxicological studies requires decisions about the number and spacing of exposure groups tested, the number of replications, the spacing of sampling times, the duration of the study, and other considerations. For example, geometric spacing of sampling times or toxicant concentrations is often used as a default design. Optimal design methods in statistics can suggest alternative spacing of sampling times that yield more precise estimates of regression coefficients. In this study, we use a computer simulation to explore the impact of the spacing of sampling times and other factors on the estimation of uptake and elimination rate constants in an experiment addressing the bioaccumulation of a contaminant. Careful selection of sampling times can result in smaller standard errors for the parameter estimates, thereby allowing the construction of smaller, more precise confidence intervals. Thus, the effort invested in constructing an optimal experimental design may result in more precise inference or in a reduction of replications in an experimental design.
Assuntos
Poluentes Ambientais/metabolismo , Modelos Biológicos , Projetos de Pesquisa , Animais , Simulação por Computador , Dinâmica não Linear , Farmacocinética , Análise de RegressãoRESUMO
The substandard nature of the housing in which most farmworkers live has detrimental effects on their health, as well as on their children's health and development. However, little research has directly documented associations between farmworker housing and health; existing research is not always comparable due to differences in design and measurement. Comparative data can help determine actual causal links between housing characteristics and farmworker health and help to evaluate the efficacy of current housing policy. The goal of this paper is to provide guidelines promoting comparable research on farmworker housing and the association of this housing with health. This paper reviews general concepts relevant to measuring farmworker housing and health, issues that should be considered in designing farmworker housing and health research, data collection methods, and measures. It concludes with recommendations for a research agenda on farmworker housing and health.
Assuntos
Coleta de Dados/métodos , Fazendeiros , Nível de Saúde , Habitação/normas , Migrantes , Agroquímicos/normas , Alérgenos , Comunicação , Aglomeração , Serviços Médicos de Emergência/métodos , Meio Ambiente , Características da Família , Inquéritos Epidemiológicos , Habitação/legislação & jurisprudência , Humanos , Projetos de Pesquisa , Características de Residência , Isolamento Social , Meios de Transporte/economia , Meios de Transporte/métodos , Estados Unidos , Abastecimento de Água/normasRESUMO
Originally shown to promote the growth and activation of B cells, thymic stromal lymphopoietin (TSLP) is now known to have wide-ranging impacts on both hematopoietic and nonhematopoietic cell lineages, including dendritic cells, basophils, eosinophils, mast cells, CD4âº, CD8⺠and natural killer T cells, B cells and epithelial cells. While TSLP's role in the promotion of TH2 responses has been extensively studied in the context of lung- and skin-specific allergic disorders, it is becoming increasingly clear that TSLP may impact multiple disease states within multiple organ systems, including the blockade of TH1/TH17 responses and the promotion of cancer and autoimmunity. This chapter will highlight recent advances in the understanding of TSLP signal transduction, as well as the role of TSLP in allergy, autoimmunity and cancer. Importantly, these insights into TSLP's multifaceted roles could potentially allow for novel therapeutic manipulations of these disorders.
Assuntos
Citocinas/metabolismo , Sistema Imunitário/metabolismo , Receptores de Citocinas/metabolismo , Transdução de Sinais , Animais , Humanos , Sistema Imunitário/imunologia , Doenças do Sistema Imunitário/imunologia , Doenças do Sistema Imunitário/metabolismo , Proteínas de Neoplasias/metabolismo , Neoplasias/imunologia , Neoplasias/metabolismo , Linfopoietina do Estroma do TimoRESUMO
Originally shown to promote the growth and activation of B cells, TSLP is now known to have wide-ranging impacts on hematopoietic and nonhematopoietic cell lineages, including DCs, basophils, eosinophils, mast cells, CD4(+), CD8(+), and NK T cells, B cells, and epithelial cells. Whereas the role of TSLP in the promotion of TH2 responses has been studied extensively in the context of lung- and skin-specific allergic disorders, it is becoming increasingly clear that TSLP may impact multiple disease states within multiple organ systems, including the blockade of TH1/TH17 responses and the promotion of cancer and autoimmunity. This review will highlight recent advances in the understanding of TSLP signal transduction, as well as the role of TSLP in allergy, autoimmunity, and cancer. Importantly, these insights into the multifaceted roles of TSLP could potentially allow for novel, therapeutic manipulations of these disorders.
Assuntos
Citocinas/imunologia , Células Dendríticas/imunologia , Hipersensibilidade/imunologia , Leucócitos/imunologia , Transdução de Sinais/imunologia , Animais , Doenças Autoimunes/imunologia , Doenças Autoimunes/patologia , Células Dendríticas/patologia , Humanos , Hipersensibilidade/patologia , Inflamação/imunologia , Inflamação/patologia , Leucócitos/patologia , Neoplasias/imunologia , Neoplasias/patologia , Linfopoietina do Estroma do TimoRESUMO
BACKGROUND: This multicenter study was undertaken to characterize the metastatic behavior of oral maxillary squamous carcinoma and to determine the role of selective neck dissection. METHODS: A retrospective, multicenter study of patients surgically treated for oral maxillary squamous carcinoma was completed. Data collected included primary tumor location, cervical lymph node status, and neck failure rate. RESULTS: The study included 146 patients. The adjusted regional metastatic rate was 31.4%. Of those N0 (clinically negative) necks treated with or without neck dissection, 14.4% developed cervical metastasis. Within the cohort, 7.5% of patients died with distant disease. The regional salvage rate was 52.9%. None of the patients with locoregional failures were salvaged. CONCLUSIONS: Maxillary palatal, alveolar, and gingival squamous carcinomas exhibit aggressive regional metastatic behavior. Surgical salvage rates for neck failure are low; therefore, selective neck dissection (levels I-III) is recommended at the time of resection of T2, T3, and T4 maxillary squamous carcinomas.
Assuntos
Carcinoma de Células Escamosas/cirurgia , Linfonodos/patologia , Linfonodos/cirurgia , Neoplasias Maxilares/cirurgia , Neoplasias Bucais/cirurgia , Esvaziamento Cervical/estatística & dados numéricos , Recidiva Local de Neoplasia/patologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Carcinoma de Células Escamosas/mortalidade , Carcinoma de Células Escamosas/patologia , Carcinoma de Células Escamosas/secundário , Estudos de Coortes , Intervalo Livre de Doença , Feminino , Humanos , Excisão de Linfonodo/métodos , Metástase Linfática , Masculino , Neoplasias Maxilares/mortalidade , Neoplasias Maxilares/patologia , Pessoa de Meia-Idade , Neoplasias Bucais/mortalidade , Neoplasias Bucais/patologia , Invasividade Neoplásica/patologia , Recidiva Local de Neoplasia/mortalidade , Recidiva Local de Neoplasia/cirurgia , Estadiamento de Neoplasias , Prognóstico , Estudos Retrospectivos , Medição de Risco , Análise de Sobrevida , Resultado do TratamentoRESUMO
Autophagy, an ancient cellular response where autophagic vacuoles are formed within the cytosol, is induced in response to a variety of cellular insults, including growth factor or nutrient withdrawal, organelle damage, and misfolded proteins. Autophagy is rapidly induced in T lymphocytes following antigenic stimulation and blockade of autophagic signaling greatly reduces T cell clonal expansion, suggesting that autophagy is primarily involved in promoting T cell survival. In contrast, a recently identified negative feedback loop involving FADD and caspase-8 limits the level of autophagy in T cells. Failure to activate caspase-8 during T cell mitogenesis leads to hyperactive autophagy and cellular death through a programmed necrotic mechanism. These findings suggest that crosstalk between these cellular processes is essential for T cell activation and homeostasis.
Assuntos
Imunidade Adaptativa/imunologia , Autofagia/imunologia , Ativação Linfocitária/imunologia , Linfócitos T , Animais , Apoptose , Caspase 8/metabolismo , Proteína de Domínio de Morte Associada a Fas/metabolismo , Homeostase , Humanos , Transdução de Sinais , Linfócitos T/citologia , Linfócitos T/imunologiaRESUMO
During an immune response, specific recognition of microbial and tumor antigens leads to the rapid proliferation of lymphocytes. Once the immunological challenge is eliminated, the vast majority of these lymphocytes must be removed via apoptosis. Cell death is also vital for the deletion of autoreactive or chronically activated lymphocytes to prevent the development of autoimmunity in the host. Such processes are highly dependent on death receptors (DRs), molecules of the TNF receptor family. While these DRs promote apoptosis, interference with DR signaling paradoxically interferes with rapid lymphocyte proliferation. Recently, we discovered that T cells lacking Fas-Associated protein with Death Domain (FADD) or caspase-8 (casp8) function, both essential for DR-induced apoptosis, succumb to hyperactivation of autophagy and die through a nonapoptotic form of cell death rather than proliferating after mitogen stimulation. We observed recruitment of FADD, casp8 and serine/threonine kinase RIPK1 to complexes containing Atg5, Atg12 and Atg16L, suggesting that the generation of early autophagosomes leads to the assembly of complexes that activate casp8. Because blockade of RIPK1 or interference with autophagic signaling inhibited this alternative death process, we propose that hyperactive autophagy induced in the absence of caspase activity leads to a necrosis-like form of death that depends on RIPK1 enzymatic function. Herein, we summarize these findings and speculate on the significance and means by which autophagy is normally activated in proliferating lymphocytes.
Assuntos
Apoptose , Autofagia , Proteína de Domínio de Morte Associada a Fas/metabolismo , Linfócitos T/citologia , Animais , Humanos , Camundongos , Modelos Biológicos , Receptores de Antígenos de Linfócitos T/metabolismo , Transdução de SinaisRESUMO
Fas-associated death domain protein (FADD) constitutes an essential component of TNFR-induced apoptotic signaling. Paradoxically, FADD has also been shown to be crucial for lymphocyte development and activation. In this study, we report that FADD is necessary for long-term maintenance of S6 kinase (S6K) activity. S6 phosphorylation at serines 240 and 244 was only observed after long-term stimulation of wild-type cells, roughly corresponding to the time before S-phase entry, and was poorly induced in T cells expressing a dominantly interfering form of FADD (FADDdd), viral FLIP, or possessing a deficiency in caspase-8. Defects in S6K1 phosphorylation were also observed. However, defective S6K1 phosphorylation was not a consequence of a wholesale defect in mammalian target of rapamycin function, because 4E-BP1 phosphorylation following T cell activation was unaffected by FADDdd expression. Although cyclin D3 up-regulation and retinoblastoma hypophosphorylation occurred normally in FADDdd T cells, cyclin E expression and cyclin-dependent kinase 2 activation were markedly impaired in FADDdd T cells. These results demonstrate that a FADD/caspase-8-signaling axis promotes T cell cycle progression and sustained S6K activity.
Assuntos
Caspase 8/fisiologia , Proteína de Domínio de Morte Associada a Fas/fisiologia , Interleucina-2/fisiologia , Proteínas Quinases S6 Ribossômicas/metabolismo , Fase S/imunologia , Transdução de Sinais/imunologia , Subpopulações de Linfócitos T/citologia , Subpopulações de Linfócitos T/enzimologia , Síndrome de Alstrom , Animais , Caspase 8/genética , Células Cultivadas , Quinase 2 Dependente de Ciclina/deficiência , Quinase 2 Dependente de Ciclina/metabolismo , Quinase 2 Dependente de Ciclina/fisiologia , Ativação Enzimática/genética , Ativação Enzimática/imunologia , Humanos , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Fosforilação , Proteínas Quinases S6 Ribossômicas/deficiência , Fase S/genética , Transdução de Sinais/genética , Subpopulações de Linfócitos T/imunologiaRESUMO
Recently, it has been demonstrated that stimulated T cells bearing defects in caspase-8 fail to promote nuclear shuttling of NF-kappaB complexes. Such cells display strikingly similar proliferative and survival defects as T cells lacking Fas-associated death domain protein (FADD) function. We characterized NF-kappaB signaling in T cells bearing a dominant-negative FADD transgene (FADDdd). Whereas FADDdd T cells displayed proliferative defects following activation, these were not a consequence of aberrant NF-kappaB signaling, as measured by IKK/IkappaB phosphorylation and IkappaB degradation. There were no appreciable defects in nuclear translocation of p65/Rel using ImageStream, a flow-based imaging cytometer. Pretreatment with benzyloxycarbonyl-Val-Ala-Asp-fluoromethylketone, a potent caspase inhibitor, also failed to impede canonical NF-kappaB signaling. Secretion of IL-2 and up-regulation of various activation markers occurred normally. Thus, FADD does not play an essential role in NF-kappaB activation, suggesting an alternative route by which this adaptor promotes the clonal expansion of T cells.