RESUMEN
BACKGROUND: Thymic stromal lymphopoietin (TSLP) is an epithelial-derived cytokine that is important for the development of type 2 inflammatory responses at mucosal surfaces. OBJECTIVE: In humans, TSLP level has been found to be elevated in the lungs of patients with asthma, and in mouse models, TSLP can promote type 2 airway inflammation, primarily through the activation of dendritic cells. However, the mechanisms underlying its role remain unclear. The objective of this study was to provide a mechanistic analysis of TSLP-mediated type 2 airway inflammation METHODS: To dissect the mechanisms of TSLP-mediated type 2 responses, mice were treated with TSLP and antigen to evaluate cellular immune responses. Flow cytometric analyses were used to follow responses in the airways, and conditional deletion of TSLP receptor and adoptive transfer were used to identify the cellular subsets involved in this inflammatory response. RESULTS: We showed that TSLP can directly promote TH2-cell differentiation in the lung, independent of the draining lymph nodes. We also identified a population of patrolling monocytes/interstitial macrophages (IMs) (CD11c-expressing IMs) that are both necessary and sufficient for TSLP-mediated TH2-cell differentiation and airway inflammation. TH2-cell-driven airway eosinophilia is attenuated by ablation of CD11c-expressing IMs or by selective deficiency of TSLP receptor signaling in these cells. More importantly, CD11c-expressing IMs are sufficient for the induction of acute TH2-cell responses in the lungs that is independent of dendritic cells and T-cell priming in the draining lymph nodes. CONCLUSION: These findings indicate a novel mechanistic role for TSLP and CD11c-expressing IMs in the development of acute TH2-cell-dependent allergic airway inflammation. This work also demonstrates a new role for TSLP in promoting type 2 responses directly in the lung.
Asunto(s)
Asma/inmunología , Diferenciación Celular/inmunología , Citocinas/inmunología , Pulmón/inmunología , Eosinofilia Pulmonar/inmunología , Células Th2/inmunología , Animales , Asma/patología , Modelos Animales de Enfermedad , Pulmón/patología , Macrófagos/inmunología , Macrófagos/patología , Ratones , Ratones Endogámicos BALB C , Monocitos/inmunología , Monocitos/patología , Eosinofilia Pulmonar/patología , Células Th2/patología , Linfopoyetina del Estroma TímicoRESUMEN
Inflammation induced during infection can both promote and suppress immunity. This contradiction suggests that inflammatory cytokines affect the immune system in a context-dependent manner. Here we show that nonspecific bystander inflammation conditions naive CD4(+) T cells for enhanced peripheral Foxp3 induction and reduced effector differentiation. This results in inhibition of immune responses in vivo via a Foxp3-dependent effect on antigen-specific naive CD4(+) T cell precursors. Such conditioning may have evolved to allow immunity to infection while limiting subsequent autoimmunity caused by release of self-antigens in the wake of infection. Furthermore, this phenomenon suggests a mechanistic explanation for the idea that early tuning of the immune system by infection affects the long-term quality of immune regulation.
Asunto(s)
Asma/inmunología , Autoinmunidad/inmunología , Efecto Espectador/inmunología , Linfocitos T CD4-Positivos/inmunología , Citocinas/inmunología , Diabetes Mellitus/inmunología , Factores de Transcripción Forkhead/inmunología , Inflamación , Autotolerancia/inmunología , Animales , Autoantígenos , Efecto Espectador/efectos de los fármacos , Linfocitos T CD4-Positivos/efectos de los fármacos , Línea Celular Tumoral , Citocinas/efectos de los fármacos , Citocinas/farmacología , Metilación de ADN , Modelos Animales de Enfermedad , Femenino , Citometría de Flujo , Factores de Transcripción Forkhead/genética , Perfilación de la Expresión Génica , Inductores de Interferón/farmacología , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Ratones Transgénicos , Tolerancia Periférica/inmunología , Poli I-C/farmacología , Regiones Promotoras Genéticas , Linfopoyetina del Estroma TímicoRESUMEN
Recent reports have highlighted greater complexity, plasticity, and functional diversity of mononuclear phagocytes (MPCs), including monocytes, macrophages, and dendritic cells (DCs), in our organs than previously understood. The functions and origins of MPCs resident within healthy organs, especially in the kidney, are less well understood, whereas studies suggest they play roles in disease states distinct from recruited monocytes. We developed an unbiased approach using flow cytometry to analyze MPCs residing in the normal mouse kidney, and identified five discrete subpopulations according to CD11b/CD11c expression as well as F4/80, CD103, CD14, CD16, and CD64 expression. In addition to distinct marker profiles, these subpopulations have different lineages and expression of genes involved in tissue homeostasis, including angiogenesis. Among them, the CD11b(int)CD11c(int) F4/80(high) subpopulation notably exhibited high capacity to produce a representative anti-inflammatory cytokine, IL-10. Each subpopulation had different degrees of both macrophage (phagocytosis) and DC (Ag presentation) capacities, with a tendency to promote differentiation of regulatory T cells, whereas two of these showed expression of transcription factors reported to be highly expressed by classical DCs, and proclivity to exit the kidney following stimulation with LPS. In summary, resident kidney MPCs comprise discrete subpopulations, which cannot be simply classified into the conventional entities, and they produce anti-inflammatory and tissue-homeostatic factors to differing degrees.
Asunto(s)
Riñón/citología , Riñón/inmunología , Sistema Mononuclear Fagocítico/citología , Sistema Mononuclear Fagocítico/inmunología , Animales , Diferenciación Celular/inmunología , Citometría de Flujo , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Microscopía Fluorescente , Sistema Mononuclear Fagocítico/metabolismo , Fenotipo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Linfocitos T/citología , Linfocitos T/inmunologíaRESUMEN
Naive CD4 T cells can differentiate into a number of functional subsets in response to Ag, including Foxp3(+) induced regulatory T cells (iTregs). The in vivo development and function of iTregs has been primarily demonstrated in systems involving Ag encountered systemically or delivered via the intestinal mucosa. In this study, we demonstrate that de novo Foxp3 expression in naive CD4 T cells is a critical mechanism for establishing tolerance for a tissue-restricted neo-self Ag. Naive CD4 T cells lacking a functional Foxp3 gene cannot achieve tolerance, but can be suppressed in vivo in the presence of wild type naive CD4 T cells. Exposure to nonspecific inflammation during priming undermines tolerance through impaired Foxp3 induction, suggesting that the microenvironment also has a role. These data show that de novo Foxp3 expression is an integral component of establishing and maintaining tolerance among naive peripheral CD4 T cells.
Asunto(s)
Autoantígenos/inmunología , Factores de Transcripción Forkhead/inmunología , Tolerancia Inmunológica/inmunología , Linfocitos T Reguladores/inmunología , Traslado Adoptivo , Animales , Autoantígenos/metabolismo , Femenino , Citometría de Flujo , Factores de Transcripción Forkhead/metabolismo , Inflamación/inmunología , Inflamación/metabolismo , Masculino , Ratones , Ratones Congénicos , Ratones Endogámicos BALB C , Linfocitos T/inmunología , Linfocitos T/metabolismo , Linfocitos T Reguladores/metabolismo , Linfocitos T Reguladores/trasplanteRESUMEN
Mice overexpressing the proallergic cytokine thymic stromal lymphopoietin (TSLP) in the skin develop a pathology resembling atopic dermatitis. RabGEF1, a guanine nucleotide exchange factor for Rab5 GTPase, is a negative regulator of IgE-dependent mast cell activation, and Rabgef1-/- and TSLP transgenic mice share many similar phenotypic characteristics, including elevated serum IgE levels and severe skin inflammation, with infiltrates of both lymphocytes and eosinophils. We report here that Rabgef1-/- mice also develop splenomegaly, lymphadenopathy, myeloid hyperplasia, and high levels of TSLP. Rabgef1-/-TSLPR-/- mice, which lack TSLP/TSLP receptor (TSLPR) signaling, had levels of blood neutrophils, spleen myeloid cells, and serum IL-4, IgG1, and IgE levels that were significantly reduced compared with those in Rabgef1-/-TSLPR+/+ mice. However, Rabgef1-/-TSLPR-/- mice, like Rag1- or eosinophil-deficient Rabgef1-/- mice, developed cutaneous inflammation and epidermal hyperplasia. Therefore, in Rabgef1-/- mice, TSLP/TSLPR interactions are not required for the development of epidermal hyperplasia but contribute to the striking myeloid hyperplasia and overproduction of immunoglobulins observed in these animals. Our study shows that RabGEF1 can negatively regulate TSLP production in vivo and that excessive production of TSLP contributes to many of the phenotypic abnormalities in Rabgef1-/- mice. However, the marked epidermal hyperplasia, cutaneous inflammation, and increased numbers of dermal mast cells associated with RabGEF1 deficiency can develop via a TSLPR-independent pathway, as well as in the absence of Rag1 or eosinophils.
Asunto(s)
Citocinas/inmunología , Epidermis/patología , Factores de Intercambio de Guanina Nucleótido/genética , Hiperplasia , Inmunoglobulinas/inmunología , Células Mieloides/inmunología , Células Mieloides/patología , Inmunidad Adaptativa , Animales , Linfocitos B/citología , Linfocitos B/inmunología , Citocinas/sangre , Epidermis/inmunología , Factores de Intercambio de Guanina Nucleótido/metabolismo , Inmunoglobulinas/sangre , Interleucina-4/inmunología , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Ratones Noqueados , Fenotipo , Receptores de Citocinas/genética , Receptores de Citocinas/metabolismo , Transducción de Señal/inmunología , Bazo/citología , Bazo/inmunología , Linfopoyetina del Estroma TímicoRESUMEN
Type-I IFNs (IFN-I) provide direct survival signals to T cells during Ag-driven proliferation. Because IFN-I production differs depending on the pathogen, we assessed CD8 T cell requirement for direct IFN-I signals during responses to vaccinia virus (VV), vesicular stomatitis virus (VSV), lymphocytic choriomeningitis virus (LCMV), and Listeria monocytogenes (LM) immunizations in vivo. IFN-I-receptor-deficient (IFN-IR(o)) CD8 T cells expanded 3- to 5-fold less and formed a diminished memory pool compared with wild-type (WT) CD8 T cells in response to VV, VSV, or LM. WT CD8 T cells expanded more robustly in response to LCMV-encoded Ags than to Ags encoded by the other three pathogens, and under these conditions the lack of direct IFN-I signals inhibited their expansion by approximately 100-fold. To test whether the high antigenic-load provided by LCMV caused greater expansion and greater IFN-I dependency, we primed WT and IFN-IR(o) OVA-specific OT-1 CD8 T cells with a fixed-number of OVA-peptide-pulsed dendritic cells along with adjuvant effect provided by LCMV, VV, VSV, or LM. Both WT and IFN-IR(o) OT-1 cells were recruited, proliferated, and differentiated into effectors in all the four cases. However, WT OT-1 cells expanded similarly in all four cases. IFN-IR(o) OT-1 cells expanded approximately 20-fold less than the WT OT-1 CD8 T cells when LCMV was used as adjuvant, whereas their expansion was affected only marginally when VV, VSV, or LM were used as adjuvants. Thus, innate/inflammatory signals induced by different pathogens contribute to CD8 T cell expansion and memory formation via distinct levels of IFN-I dependence.
Asunto(s)
Linfocitos T CD8-positivos/patología , Diferenciación Celular/inmunología , Memoria Inmunológica , Interferón Tipo I/fisiología , Listeriosis/patología , Coriomeningitis Linfocítica/patología , Infecciones por Rhabdoviridae/patología , Transducción de Señal/inmunología , Vaccinia/patología , Animales , Células Presentadoras de Antígenos/inmunología , Células Presentadoras de Antígenos/microbiología , Células Presentadoras de Antígenos/virología , Antígenos Bacterianos/biosíntesis , Antígenos Bacterianos/metabolismo , Antígenos Virales/biosíntesis , Antígenos Virales/metabolismo , Linfocitos T CD8-positivos/inmunología , Linfocitos T CD8-positivos/microbiología , Linfocitos T CD8-positivos/virología , Proliferación Celular , Células Clonales , Inmunidad Innata , Inflamación/inmunología , Inflamación/microbiología , Inflamación/virología , Listeriosis/inmunología , Listeriosis/microbiología , Coriomeningitis Linfocítica/inmunología , Coriomeningitis Linfocítica/virología , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Infecciones por Rhabdoviridae/inmunología , Infecciones por Rhabdoviridae/virología , Vaccinia/inmunología , Vaccinia/virologíaRESUMEN
T cell expansion and memory formation are generally more effective when elicited by live organisms than by inactivated vaccines. Elucidation of the underlying mechanisms is important for vaccination and therapeutic strategies. We show that the massive expansion of antigen-specific CD8 T cells that occurs in response to viral infection is critically dependent on the direct action of type I interferons (IFN-Is) on CD8 T cells. By examining the response to infection with lymphocytic choriomeningitis virus using IFN-I receptor-deficient (IFN-IR(0)) and -sufficient CD8 T cells adoptively transferred into normal IFN-IR wild-type hosts, we show that the lack of direct CD8 T cell contact with IFN-I causes >99% reduction in their capacity to expand and generate memory cells. The diminished expansion of IFN-IR(0) CD8 T cells was not caused by a defect in proliferation but by poor survival during the antigen-driven proliferation phase. Thus, IFN-IR signaling in CD8 T cells is critical for the generation of effector and memory cells in response to viral infection.
Asunto(s)
Infecciones por Arenaviridae/inmunología , Linfocitos T CD8-positivos/metabolismo , Memoria Inmunológica/inmunología , Interferón Tipo I/metabolismo , Activación de Linfocitos/inmunología , Virus de la Coriomeningitis Linfocítica/inmunología , Transducción de Señal/inmunología , Animales , Linfocitos T CD8-positivos/inmunología , Fluoresceínas , Interferón Tipo I/inmunología , Ratones , Ratones Endogámicos C57BL , Receptores de Interferón/genética , SuccinimidasRESUMEN
Flagellin is the structural component of flagella produced by many pathogenic bacteria and is a potent proinflammatory molecule that mediates these effects through Toll-like receptor (TLR) 5. In Listeria monocytogenes (LM), flagellin expression is regulated by temperature and has been described as being shut off at 37 degrees C. In this study, we demonstrate that TLR5-mediated cell activation and flagellin expression is maintained at 37 degrees C in some laboratory-adapted strains and in approximately 20% of LM clinical isolates. To determine the role of flagellin in LM infection, a targeted mutation in the structural gene for flagellin (flaA) was generated in a parental LM strain that expressed flagellin under all conditions examined. In vitro studies demonstrated that this deltaflaA mutant was (i). non-motile; (ii). not able to activate TLR5-transfected HeLa cells; and (iii). induced tumour necrosis factor (TNF)-alpha production in approximately 50% fewer CD11b+ cells in splenocytes from normal mice compared with the parental strain. However, there was no significant alteration in virulence of the deltaflaA mutant after either intravenous or oral murine infection. Similarly, there was no difference in the generation of LM-specific CD8 or CD4 T cells after intravenous or oral infection. These data indicate that flagellin is not essential for LM pathogenesis or for the induction of LM-specific adaptive immune responses in normal mice.
Asunto(s)
Flagelina/inmunología , Listeria monocytogenes/inmunología , Listeriosis/inmunología , Animales , Modelos Animales de Enfermedad , Flagelina/genética , Flagelina/metabolismo , Listeria monocytogenes/metabolismo , Listeria monocytogenes/fisiología , RatonesRESUMEN
TGF-beta opposes proliferative signaling by IL-2 through mechanisms that remain incompletely defined. In a well-characterized CD8(+) T cell model using wild-type and mutated IL-2 receptors, we examined the effects of TGF-beta on distinct IL-2 signaling events in CD8(+) T cells. IL-2 induces c-myc, cyclin D2, and cyclin E in a redundant manner through the Shc and STAT5 pathways. TGF-beta inhibited the ability of either the Shc or STAT5 pathway to induce these genes, as well as T cell proliferation. The inhibitory effects of TGF-beta were reversed by expression of a dominant-negative form of Smad3. TGF-beta did not impair proximal signaling by Shc or STAT5, and induction of some downstream genes, including cytokine-inducible Src homology-2-containing protein (CIS), bcl-x(L), and bcl-2, was spared. Experiments with c-fos, cyclin D2, and CIS reporter genes revealed that promoter-proximal regulatory elements dictate the sensitivity of IL-2 target genes to inhibition by TGF-beta. By leaving the Shc and STAT5 pathways functional while inhibiting their target genes selectively, TGF-beta was found to uncouple the proliferative and antiapoptotic functions of IL-2. Thus, TGF-beta is not a simple antagonist of IL-2, but rather serves to qualitatively modify the IL-2 signal to create a unique pattern of gene expression that neither cytokine can induce independently.
Asunto(s)
Apoptosis/inmunología , Proteínas de Unión al ADN/fisiología , Interleucina-2/fisiología , Proteínas de la Leche , Mitógenos/fisiología , Transducción de Señal/inmunología , Transactivadores/fisiología , Factor de Crecimiento Transformador beta/fisiología , Animales , Apoptosis/genética , División Celular/inmunología , Línea Celular , Supervivencia Celular/inmunología , Proteínas de Unión al ADN/antagonistas & inhibidores , Regulación de la Expresión Génica/inmunología , Genes Reporteros/inmunología , Inhibidores de Crecimiento/fisiología , Interleucina-2/antagonistas & inhibidores , Ratones , Mitógenos/antagonistas & inhibidores , Mitógenos/genética , Regiones Promotoras Genéticas/inmunología , Receptores de Interleucina-2/antagonistas & inhibidores , Receptores de Interleucina-2/genética , Receptores de Interleucina-2/fisiología , Secuencias Reguladoras de Ácidos Nucleicos/inmunología , Factor de Transcripción STAT5 , Transducción de Señal/genética , Proteínas Smad , Proteína smad3 , Linfocitos T/citología , Linfocitos T/inmunología , Linfocitos T/metabolismo , Transactivadores/antagonistas & inhibidores , Dominios Homologos src/genética , Dominios Homologos src/inmunologíaRESUMEN
Diacylglycerol-dependent signaling plays an important role in signal transduction through T- and B-lymphocyte antigen receptors. Recently, a novel serine-threonine kinase of the protein kinase C (PKC) family has been described and designated as PKCnu. PKCnu has two putative diacylglycerol binding C1 domains, suggesting that it may participate in a novel diacylglycerol-mediated signaling pathway. Here we show that both endogenous and recombinant PKCnu are trans-located to the plasma membrane and activated by the diacylglycerol mimic phorbol 12-myristate 13-acetate. Mutational analysis demonstrates that PKCnu activation is dependent on trans-phosphorylation of two conserved activation loop serine residues. We also find that PKCnu is an important physiologic target of the B-cell receptor (BCR), because PKCnu is found to be abundantly expressed in chicken and human B-cell lines and, in addition, exhibits robust activation after BCR engagement. Genetic and pharmacologic analyses of BCR-mediated PKCnu activation indicate that it requires intact phospholipase Cgamma and PKC signaling pathways. Furthermore, in co-transfection assays, PKCnu can be trans-phosphorylated by the novel PKC isozymes PKCepsilon, PKCeta, or PKCtheta but not the classical PKC enzyme, PKCalpha. Taken together, these results suggest that PKCnu is an important component of signaling pathways downstream from novel PKC enzymes after B-cell receptor engagement.