RESUMEN
T helper 17 (Th17) cell development is driven by cytokines including transforming growth factor-ß (TGF-ß), interleukin-6 (IL-6), IL-1, and IL-23. Regulatory T (Treg) cells can provide the TGF-ß in vitro, but their role in vivo remains unclear, particularly because Treg cells inhibit inflammation in many models of Th17 cell-associated autoimmunity. We used mice expressing Diphtheria toxin receptor under control of the Foxp3 promoter to deplete Foxp3(+) Treg cells in adult mice during in vivo Th17 cell priming. Treg cell depletion resulted in a reduced frequency of antigen-specific IL-17 producers in draining lymph nodes and blood, correlating with reduced inflammatory skin responses. In contrast, Treg cells did not promote IL-17 secretion after initial activation stages. Treg cell production of TGF-ß was not required for Th17 cell promotion, and neither was suppression of Th1 cell-associated cytokines. Rather, regulation of IL-2 availability and resultant signaling through CD25 by Treg cells was found to play an important role.
Asunto(s)
Diferenciación Celular , Factores de Transcripción Forkhead/inmunología , Interleucina-17/inmunología , Interleucina-2/inmunología , Linfocitos T Colaboradores-Inductores/citología , Linfocitos T Colaboradores-Inductores/inmunología , Linfocitos T Reguladores/inmunología , Animales , Células Cultivadas , Ratones , Ratones Transgénicos , Modelos InmunológicosRESUMEN
Dendritic cells (DCs) as professional antigen-presenting cells play an important role in the initiation and modulation of the adaptive immune response. However, their role in the innate immune response against bacterial infections is not completely defined. Here we have analyzed the role of DCs and their impact on the innate anti-bacterial host defense in an experimental infection model of Yersinia enterocolitica (Ye). We used CD11c-diphtheria toxin (DT) mice to deplete DCs prior to severe infection with Ye. DC depletion significantly increased animal survival after Ye infection. The bacterial load in the spleen of DC-depleted mice was significantly lower than that of control mice throughout the infection. DC depletion was accompanied by an increase in the serum levels of CXCL1, G-CSF, IL-1α, and CCL2 and an increase in the numbers of splenic phagocytes. Functionally, splenocytes from DC-depleted mice exhibited an increased bacterial killing capacity compared to splenocytes from control mice. Cellular studies further showed that this was due to an increased production of reactive oxygen species (ROS) by neutrophils. Adoptive transfer of neutrophils from DC-depleted mice into control mice prior to Ye infection reduced the bacterial load to the level of Ye-infected DC-depleted mice, suggesting that the increased number of phagocytes with additional ROS production account for the decreased bacterial load. Furthermore, after incubation with serum from DC-depleted mice splenocytes from control mice increased their bacterial killing capacity, most likely due to enhanced ROS production by neutrophils, indicating that serum factors from DC-depleted mice account for this effect. In summary, we could show that DC depletion triggers phagocyte accumulation in the spleen and enhances their anti-bacterial killing capacity upon bacterial infection.
Asunto(s)
Células Dendríticas/patología , Inmunidad Innata/fisiología , Fagocitos/fisiología , Yersiniosis/inmunología , Yersinia enterocolitica/inmunología , Traslado Adoptivo , Animales , Bacterias/inmunología , Separación Celular , Células Cultivadas , Femenino , Homeostasis/inmunología , Ratones , Ratones Transgénicos , Neutrófilos/trasplante , Fagocitos/inmunología , Regulación hacia Arriba/inmunología , Yersiniosis/patología , Yersiniosis/terapiaRESUMEN
Gastrointestinal helminth infections are extremely prevalent in many human populations and are associated with downmodulated immune responsiveness. In the experimental model system of Heligmosomoides polygyrus, a chronic infection establishes in mice, accompanied by a modulated Th2 response and increased regulatory T cell (Treg) activity. To determine if dendritic cell (DC) populations in the lymph nodes draining the intestine are responsible for the regulatory effects of chronic infection, we first identified a population of CD11c(lo) nonplasmacytoid DCs that expand after chronic H. polygyrus infection. The CD11c(lo) DCs are underrepresented in magnetic bead-sorted preparations and spared from deletion in CD11c-diptheria toxin receptor mice. After infection, CD11c(lo) DCs did not express CD8, CD103, PDCA, or Siglec-H and were poorly responsive to TLR stimuli. In DC/T cell cocultures, CD11c(lo) DCs from naive and H. polygyrus-infected mice could process and present protein Ag, but induced lower levels of Ag-specific CD4(+) T cell proliferation and effector cytokine production, and generated higher percentages of Foxp3(+) T cells in the presence of TGF-ß. Treg generation was also dependent on retinoic acid receptor signaling. In vivo, depletion of CD11c(hi) DCs further favored the dominance of the CD11c(lo) DC phenotype. After CD11c(hi) DC depletion, effector responses were inhibited dramatically, but the expansion in Treg numbers after H. polygyrus infection was barely compromised, showing a significantly higher regulatory/effector CD4(+) T cell ratio compared with that of CD11c(hi) DC-intact animals. Thus, the proregulatory environment of chronic intestinal helminth infection is associated with the in vivo predominance of a newly defined phenotype of CD11c(lo) tolerogenic DCs.
Asunto(s)
Células Dendríticas/inmunología , Helmintiasis/inmunología , Parasitosis Intestinales/inmunología , Animales , Antígenos CD/inmunología , Antígenos CD11/inmunología , Recuento de Células , Enfermedad Crónica , Inmunidad , Inmunofenotipificación , Cadenas alfa de Integrinas/inmunología , Ratones , Nematospiroides dubius/inmunología , Linfocitos T Reguladores/inmunologíaRESUMEN
Dendritic cells (DCs) are key components of the adaptive immune system contributing to initiation and regulation of T cell responses. T cells continuously scan DCs in lymphoid organs for the presence of foreign antigen. However, little is known about the functional consequences of these frequent T cell-DC interactions without cognate antigen. Here we demonstrate that these contacts in the absence of foreign antigen serve an important function, namely, induction of a basal activation level in T cells required for responsiveness to subsequent encounters with foreign antigens. This basal activation is provided by self-recognition of MHC molecules on DCs. Following DC depletion in mice, T cells became impaired in TCR signaling and immune synapse formation, and consequently were hyporesponsive to antigen. This process was reversible, as T cells quickly recovered when the number of DCs returned to a normal level. The extent of T cell reactivity correlated with the degree of DC depletion in lymphoid organs, suggesting that a full DC compartment guarantees optimal T cell responsiveness. These findings indicate that DCs are specialized cells that not only present foreign antigen, but also promote a "tonic" state in T cells for antigen responsiveness.
Asunto(s)
Células Dendríticas/inmunología , Linfocitos T/inmunología , Animales , Presentación de Antígeno , Linfocitos T CD4-Positivos/citología , Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD8-positivos/citología , Linfocitos T CD8-positivos/inmunología , Recuento de Células , Supervivencia Celular/inmunología , Células Dendríticas/citología , Factor de Crecimiento Similar a EGF de Unión a Heparina , Humanos , Sinapsis Inmunológicas/inmunología , Péptidos y Proteínas de Señalización Intercelular/genética , Péptidos y Proteínas de Señalización Intercelular/inmunología , Activación de Linfocitos , Ratones , Ratones Congénicos , Ratones Endogámicos C57BL , Ratones Transgénicos , Receptores de Antígenos de Linfocitos T/metabolismo , Proteínas Recombinantes/genética , Proteínas Recombinantes/inmunología , Transducción de Señal/inmunología , Linfocitos T/citología , Quimera por Trasplante/inmunologíaRESUMEN
Foxp3(+)CD4(+) regulatory T cells (Tregs) are crucial in maintaining self-tolerance and limiting immune responses to pathogens. Shifting the sensitive balance between Tregs and effector T cells requires extensive knowledge of the homeostatic properties of the different T cell populations. For the investigation of Treg homeostatic expansion, we introduce in this study novel BAC transgenic mice, designated Foxp3.LuciDTR, coexpressing enhanced GFP, luciferase for bioluminescence imaging of Tregs, and the diphtheria toxin receptor (DTR) for specific ablation of Tregs. Of several founder lines, Foxp3.LuciDTR-4 mice displayed approximately 95% Treg depletion following injection of DT, resulting in activation of conventional CD4(+) T cells, probably due to lack of control by Tregs. In contrast, Foxp3.LuciDTR-3 mice displayed only approximately 70% Treg depletion without concomitant activation of CD4(+) T cells and represented, therefore, a suitable model to study Treg homeostasis in an environment where other T cell populations were not altered. After depletion, the Treg compartment recovered to its original size in approximately 2 wk. This recovery was mediated in a thymus-independent fashion by homeostatic proliferation of the surviving, nondepleted Tregs. The proliferating Tregs acquired an activated phenotype and maintained their suppressive capacity. Studies involving DT-mediated depletion of dendritic cells in CD11c.DOG mice showed that dendritic cells were required for optimal Treg homeostasis. In addition, IL-2 was identified as an essential factor for homeostatic recovery of the Treg compartment. These results show that Treg homeostasis is specifically regulated by the size of the Treg compartment and is independent of proliferation of conventional T cells.
Asunto(s)
Proliferación Celular , Células Dendríticas/inmunología , Factores de Transcripción Forkhead/biosíntesis , Homeostasis/inmunología , Linfocitos T Reguladores/citología , Linfocitos T Reguladores/inmunología , Animales , Toxina Diftérica/genética , Factores de Transcripción Forkhead/genética , Homeostasis/genética , Proteínas Luminiscentes/genética , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Ratones Transgénicos , Linfocitos T Reguladores/metabolismoRESUMEN
Dendritic cells (DCs) are key coordinators of the immune response, governing the choice between tolerance and immunity. Despite their importance, the mechanisms controlling the size of the DC compartment are largely unknown. Using a mouse model allowing continuous DC depletion, we show that maintenance of DC numbers in spleen is an active process mediated by Flt3-L-dependent regulation of precursor differentiation into DCs, rather than by changes in proliferation of the differentiated DCs. In particular, the frequency and differentiation potential of intrasplenic DC precursors increased in response to reduced DC numbers. Levels of Flt3-L, a cytokine required for DC differentiation, increased in the blood after DC depletion and returned to normal levels once the DC compartment filled up again. Our data suggest a feedback regulation of DC homeostasis whereby reduction of the DC pool size promotes differentiation of their precursors, via increased Flt3-L availability. This mechanism is different to those known for other immune cell types, such as the B- and T-cell compartments, whereby lymphopenia induces proliferation of already differentiated lymphocytes.
Asunto(s)
Diferenciación Celular/inmunología , Células Dendríticas/citología , Retroalimentación Fisiológica/fisiología , Homeostasis/inmunología , Células Madre/citología , Traslado Adoptivo , Animales , Células Dendríticas/inmunología , Ensayo de Inmunoadsorción Enzimática , Citometría de Flujo , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Bazo/citología , Bazo/inmunología , Células Madre/inmunología , Tirosina Quinasa 3 Similar a fmsRESUMEN
Dysregulation of T lymphocyte function underpins the development of autoimmune and allergic diseases. These autoantigen-, or allergen-reactive pathogenic T cells are rare within the entire immune repertoire and it is therefore desirable to develop more specific therapies than are currently in use to directly target these cells and avoid adverse side effects. The obvious approach is to use the antigens that are recognized to impose a state of T cell tolerance. T cells recognize antigens as peptide fragments and we can therefore produce the relevant antigens as synthetic peptides. It has been known for many years that the decision of the T cell to mount a productive response (immunity) or to remain silent (tolerance) is controlled by the form in which the antigen is administered. Antigen with adjuvant leads to immunity, whereas soluble antigen without adjuvant leads to tolerance. This paradigm has been used successfully to induce tolerance with soluble peptides, preventing several animal models of autoimmune and allergic disease. These findings obviously have exciting potential for translation to human diseases. However, the basic immune mechanisms that lead to tolerance versus immunity are only beginning to be unravelled. The "effector" phase of tolerance also remains controversial with evidence for T cell death, anergy and the development of immunoregulatory function. This latter possibility of specifically generating autoantigen- or allergen-reactive regulatory T cells is particularly attractive. Here we review recent advances in our understanding of the requirements for tolerance induction and the potential for establishing dominant immune-regulation with peptide therapy.
Asunto(s)
Presentación de Antígeno , Tolerancia Inmunológica , Péptidos/uso terapéutico , Linfocitos T/fisiología , Animales , Antígenos CD/genética , Antígenos CD/metabolismo , Antígenos de Diferenciación/genética , Antígenos de Diferenciación/metabolismo , Antígenos CD28/genética , Antígenos CD28/metabolismo , Antígenos CD40/genética , Antígenos CD40/metabolismo , Ligando de CD40/metabolismo , Antígeno CTLA-4 , Células Dendríticas/inmunología , Humanos , Inmunidad Celular , Ratones , Fenotipo , Transducción de Señal , Linfocitos T/trasplante , Linfocitos T Reguladores/efectos de los fármacosRESUMEN
Although dendritic cells (DCs) are adept initiators of CD4(+) T cell responses, their fundamental importance in this regard in Th2 settings remains to be demonstrated. We have used CD11c-diphtheria toxin (DTx) receptor mice to deplete CD11c(+) cells during the priming stage of the CD4(+) Th2 response against the parasitic helminth Schistosoma mansoni. DTx treatment significantly depleted CD11c(+) DCs from all tissues tested, with 70-80% efficacy. Even this incomplete depletion resulted in dramatically impaired CD4(+) T cell production of Th2 cytokines, altering the balance of the immune response and causing a shift toward IFN-γ production. In contrast, basophil depletion using Mar-1 antibody had no measurable effect on Th2 induction in this system. These data underline the vital role that CD11c(+) antigen-presenting cells can play in orchestrating Th2 development against helminth infection in vivo, a response that is ordinarily balanced so as to prevent the potentially damaging production of inflammatory cytokines.
Asunto(s)
Presentación de Antígeno , Antígeno CD11c/inmunología , Células Dendríticas/inmunología , Esquistosomiasis mansoni/inmunología , Células Th2/inmunología , Animales , Basófilos/inmunología , Antígeno CD11c/genética , Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD4-Positivos/metabolismo , Factor de Crecimiento Similar a EGF de Unión a Heparina , Humanos , Péptidos y Proteínas de Señalización Intercelular/genética , Péptidos y Proteínas de Señalización Intercelular/inmunología , Interferón gamma/biosíntesis , Interferón gamma/inmunología , Procedimientos de Reducción del Leucocitos , Activación de Linfocitos , Ratones , Schistosoma mansoni/inmunologíaRESUMEN
Dendritic cells (DC) are known to support the activation of natural killer (NK) cells. However, little is known about the role for DC in NK-cell homeostasis. In order to investigate this question, a novel bacterial artificial chromosome transgenic mouse model was generated in which the diphtheria toxin receptor is expressed under the CD11c promoter. In these mice efficient DC depletion can be achieved over prolonged periods of time by multiple injections of diphtheria toxin. We show here that NK cells require DC for full acquisition of effector function in vivo in response to the bacterial-derived TLR ligand CpG. Importantly, DC were found to play an instrumental role for maintaining normal homeostasis of NK cells. This is achieved by IL-15 production by DC, which supports the homeostatic proliferation of NK cells.
Asunto(s)
Antígeno CD11c/inmunología , Células Dendríticas/inmunología , Interleucina-15/inmunología , Células Asesinas Naturales/inmunología , Animales , Antígeno CD11c/genética , Proliferación Celular , Cromosomas Artificiales Bacterianos , Células Dendríticas/metabolismo , Toxina Diftérica/farmacología , Interleucina-15/biosíntesis , Células Asesinas Naturales/metabolismo , Activación de Linfocitos , Ratones , Ratones Endogámicos C57BL , Ratones TransgénicosRESUMEN
An adjuvant can be defined as an agent that non-specifically promotes the immune response to an accompanying antigen. Ligation of CD40 on the surface of the antigen-presenting cell leads to upregulation of OX40 ligand which, in turn, ligates OX40 on the activated T cell resulting in prolonged T cell proliferation/survival, boosting the immune response. Thus agonistic anti-CD40 and anti-OX40 might be viewed as "adjuvant antibodies" and have been shown in diverse experimental systems to either boost immune responses or prevent the establishment of immunological tolerance. Here we describe that both these antibodies are able to prevent the induction of tolerance induced using soluble peptide antigen. However, unlike lipopolysaccharide, they are not sufficient to convert tolerance to immunity (i.e. they are not true adjuvants in this system). Using mice that are prone to either Th1 or Th2 immunity under identical immunization conditions, we show that the effects of anti-OX40 are quantitative -- boosting whichever response is dominant. In contrast, anti-CD40 boosts Th1 immunity and converts a Th2 response to Th1. We conclude that, although these two antibodies seem to impact on the same molecular pathway of costimulation to prevent tolerance, their effects are qualitatively distinct and their use cannot be viewed as interchangeable.
Asunto(s)
Adyuvantes Inmunológicos/farmacología , Anticuerpos Monoclonales/farmacología , Antígenos de Diferenciación/inmunología , Antígenos CD40/inmunología , Tolerancia Inmunológica/efectos de los fármacos , Células TH1/inmunología , Células Th2/inmunología , Animales , Anticuerpos Monoclonales/inmunología , Células Presentadoras de Antígenos/citología , Células Presentadoras de Antígenos/inmunología , Antígenos de Diferenciación/genética , Antígenos CD40/genética , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Supervivencia Celular/inmunología , Células Cultivadas , Tolerancia Inmunológica/inmunología , Glicoproteínas de Membrana/inmunología , Ratones , Ratones Noqueados , Ligando OX40 , Especificidad de la Especie , Células TH1/citología , Células Th2/citología , Factores de Necrosis Tumoral/inmunologíaRESUMEN
Steady-state dendritic cells (DC) present peptide-MHC complexes to T cells in a tolerogenic manner, presumably because of deficient costimulation. However, it is clear that the path to tolerance involves initial T cell activation, suggesting that the deficit may lie in late-acting costimulatory molecules. With this in mind we have investigated the kinetics of expression of several costimulatory pairs on DC and OVA-reactive T cells after i.v. injection of mice with peptide and LPS (immunity), or peptide alone (tolerance). We find that T cells up-regulate CD154, OX40, RANKL and PD-1 whether they are destined for tolerance or immunity, although there are some differences in the levels and length of expression. In contrast, when analyzing DC, we found that up-regulation of CD80, CD86, CD40, RANK and PDL-1 occurred only when peptide was co-administered with LPS. These data give a picture of the T cell looking for costimulatory cues that are not forthcoming when pMHC is presented by steady-state DC, leading to tolerance. However, we did see a strong and rapid up-regulation of RANKL on T cells that occurred specifically when peptide was given in the absence of LPS, suggesting a possible positive signal influencing the decision between tolerance and immunity.
Asunto(s)
Células Dendríticas/inmunología , Sistema Inmunológico/fisiología , Tolerancia Inmunológica/inmunología , Linfocitos T/inmunología , Animales , Antígenos CD/inmunología , Antígeno B7-1/inmunología , Antígeno B7-2 , Antígenos CD40/inmunología , Ligando de CD40/inmunología , Proteínas Portadoras/inmunología , Sistema Inmunológico/inmunología , Interleucina-2/biosíntesis , Interleucina-2/genética , Interleucina-2/inmunología , Cinética , Glicoproteínas de Membrana/inmunología , Ratones , Ratones Endogámicos C57BL , Ligando RANK , Receptor Activador del Factor Nuclear kappa-BRESUMEN
The decision to mount a T cell response to antigen (Ag) is dependent on the cellular context in which the Ag is presented. Activated dendritic cells (DC) are potent stimulators of immune responses, an ability which is linked to their high expression of several costimulatory molecules. In contrast, resting DC have been implicated in the generation of self tolerance, presumably due to their reduced costimulatory capacity. However, the precise molecular basis for the choice between Ag-induced immunity and unresponsiveness remains unclear. We show here that CD40 plays an important rolein this decision. Systemic administration of Ag-loaded, CD40-deficient DC failed to induce a productive primary T cell expansion and rendered mice relatively unresponsive to subsequent immunization with Ag in adjuvant. Using a TCR-transgenic T cell transfer system, we found that CD40(-/-) DC triggered an initial T cell activation that could not be sustained, resulting in loss of Ag-reactive T cells and reduced cytokine production by those T cells that did persist. Furthermore, administration of CD40(-/-) DC that had been loaded with a central nervous system autoantigen was found to protect mice from autoimmune pathology. These data implicate the CD40:CD40L interaction as a key checkpoint in the development of T cell immunity rather than tolerance.