RESUMEN
Mast cell (MC) mediator release after crosslinking of surface-bound IgE antibody by ingested antigen underlies food allergy. However, IgE antibodies are not uniformly associated with food allergy, and intestinal MC load is an important determinant. Atopic dermatitis (AD), characterized by pruritis and cutaneous sensitization to allergens, including foods, is strongly associated with food allergy. Tape stripping mouse skin, a surrogate for scratching, caused expansion and activation of small intestinal MCs, increased intestinal permeability, and promoted food anaphylaxis in sensitized mice. Tape stripping caused keratinocytes to systemically release interleukin-33 (IL-33), which synergized with intestinal tuft-cell-derived IL-25 to drive the expansion and activation of intestinal type-2 innate lymphoid cells (ILC2s). These provided IL-4, which targeted MCs to expand in the intestine. Duodenal MCs were expanded in AD. In addition to promoting cutaneous sensitization to foods, scratching may promote food anaphylaxis in AD by expanding and activating intestinal MCs.
Asunto(s)
Dermatitis Atópica/inmunología , Hipersensibilidad a los Alimentos/inmunología , Mucosa Intestinal/inmunología , Linfocitos/inmunología , Mastocitos/inmunología , Adolescente , Anafilaxia/inmunología , Animales , Proliferación Celular , Niño , Preescolar , Femenino , Humanos , Inmunoglobulina E/inmunología , Interleucina-13/metabolismo , Interleucina-33/metabolismo , Interleucina-4/metabolismo , Interleucinas/metabolismo , Mucosa Intestinal/citología , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Transducción de Señal/inmunología , Piel/inmunología , Piel/lesionesRESUMEN
CD4+ T cells are central mediators of autoimmune pathology; however, defining their key effector functions in specific autoimmune diseases remains challenging. Pathogenic CD4+ T cells within affected tissues may be identified by expression of markers of recent activation. Here we use mass cytometry to analyse activated T cells in joint tissue from patients with rheumatoid arthritis, a chronic immune-mediated arthritis that affects up to 1% of the population. This approach revealed a markedly expanded population of PD-1hiCXCR5-CD4+ T cells in synovium of patients with rheumatoid arthritis. However, these cells are not exhausted, despite high PD-1 expression. Rather, using multidimensional cytometry, transcriptomics, and functional assays, we define a population of PD-1hiCXCR5- 'peripheral helper' T (TPH) cells that express factors enabling B-cell help, including IL-21, CXCL13, ICOS, and MAF. Like PD-1hiCXCR5+ T follicular helper cells, TPH cells induce plasma cell differentiation in vitro through IL-21 secretion and SLAMF5 interaction (refs 3, 4). However, global transcriptomics highlight differences between TPH cells and T follicular helper cells, including altered expression of BCL6 and BLIMP1 and unique expression of chemokine receptors that direct migration to inflamed sites, such as CCR2, CX3CR1, and CCR5, in TPH cells. TPH cells appear to be uniquely poised to promote B-cell responses and antibody production within pathologically inflamed non-lymphoid tissues.
Asunto(s)
Artritis Reumatoide/inmunología , Artritis Reumatoide/patología , Linfocitos B/inmunología , Linfocitos T Colaboradores-Inductores/inmunología , Linfocitos T Colaboradores-Inductores/patología , Artritis Reumatoide/sangre , Linfocitos B/patología , Diferenciación Celular , Movimiento Celular , Quimiocina CXCL13/metabolismo , Perfilación de la Expresión Génica , Humanos , Proteína Coestimuladora de Linfocitos T Inducibles/metabolismo , Interleucinas/metabolismo , Factores Activadores de Macrófagos , Factor 1 de Unión al Dominio 1 de Regulación Positiva , Receptor de Muerte Celular Programada 1/metabolismo , Proteínas Proto-Oncogénicas c-bcl-6/metabolismo , Receptores CXCR5/deficiencia , Receptores CXCR5/metabolismo , Receptores de Quimiocina/metabolismo , Proteínas Represoras/metabolismo , Familia de Moléculas Señalizadoras de la Activación Linfocitaria/metabolismo , Líquido Sinovial/inmunología , Linfocitos T Colaboradores-Inductores/metabolismoRESUMEN
The role of stromal fibroblasts in chronic inflammation is unfolding. In rheumatoid arthritis, leukocyte-derived cytokines TNF and IL-17A work together, activating fibroblasts to become a dominant source of the hallmark cytokine IL-6. However, IL-17A alone has minimal effect on fibroblasts. To identify key mediators of the synergistic response to TNF and IL-17A in human synovial fibroblasts, we performed time series, dose-response, and gene-silencing transcriptomics experiments. Here we show that in combination with TNF, IL-17A selectively induces a specific set of genes mediated by factors including cut-like homeobox 1 (CUX1) and IκBζ (NFKBIZ). In the promoters of CXCL1, CXCL2, and CXCL3, we found a putative CUX1-NF-κB binding motif not found elsewhere in the genome. CUX1 and NF-κB p65 mediate transcription of these genes independent of LIFR, STAT3, STAT4, and ELF3. Transcription of NFKBIZ, encoding the atypical IκB factor IκBζ, is IL-17A dose-dependent, and IκBζ only mediates the transcriptional response to TNF and IL-17A, but not to TNF alone. In fibroblasts, IL-17A response depends on CUX1 and IκBζ to engage the NF-κB complex to produce chemoattractants for neutrophil and monocyte recruitment.
Asunto(s)
Proteínas Adaptadoras Transductoras de Señales/metabolismo , Artritis Reumatoide/metabolismo , Fibroblastos/metabolismo , Proteínas de Homeodominio/metabolismo , Inflamación/metabolismo , Interleucina-17/fisiología , Proteínas Represoras/metabolismo , Factores de Transcripción/metabolismo , Transcriptoma/fisiología , Factor de Necrosis Tumoral alfa/fisiología , Proteínas Adaptadoras Transductoras de Señales/genética , Artritis Reumatoide/genética , Células Cultivadas , Quimiocina CXCL1/genética , Quimiocina CXCL2/genética , Quimiocinas CXC/genética , Factores Quimiotácticos/genética , Fibroblastos/efectos de los fármacos , Proteínas de Homeodominio/genética , Humanos , Inflamación/genética , Interleucina-17/farmacología , Interleucina-6/genética , Metaloproteinasa 3 de la Matriz/metabolismo , Monocitos/efectos de los fármacos , Monocitos/fisiología , ARN Interferente Pequeño/genética , Proteínas Represoras/genética , Células del Estroma/efectos de los fármacos , Células del Estroma/metabolismo , Líquido Sinovial , Factor de Transcripción ReIA/metabolismo , Factores de Transcripción/genética , Transcriptoma/efectos de la radiación , Factor de Necrosis Tumoral alfa/farmacologíaRESUMEN
Mast cells (MCs) are constitutively present in most tissues and a distinct subset of MCs can also be induced upon host responses to inflammation. The hematopoietic lineage development of tissue MCs is unique compared to other myeloid-derived cells because it is early lineage progenitors, undetectable by histochemistry, that leave the bone marrow to enter the circulation. These immature lineage MCs immediately undergo transendothelial recruitment into peripheral tissues wherein the appearance of secretory granules with a particular protease phenotype is regulated by the peripheral tissue. In this Perspective, we discuss our current understanding of how these unique immunocytes arise, traffic to various sites, and may or may not mature into tissue-directed granulated phenotypes and query whether a granulated end stage is their only intended role.
Asunto(s)
Mastocitos/inmunología , Mastocitos/metabolismo , Animales , Biomarcadores/metabolismo , Diferenciación Celular , Linaje de la Célula , Células Madre Hematopoyéticas/citología , Humanos , Péptido Hidrolasas/metabolismoRESUMEN
Mast cells (MCs) are tissue resident sentinels that mature and orchestrate inflammation in response to infection and allergy. While they are also frequently observed in tumors, the contribution of MCs to carcinogenesis remains unclear. Here, we show that sequential oncogenic events in gut epithelia expand different types of MCs in a temporal-, spatial-, and cytokine-dependent manner. The first wave of MCs expands focally in benign adenomatous polyps, which have elevated levels of IL-10, IL-13, and IL-33, and are rich in type-2 innate lymphoid cells (ILC2s). These vanguard MCs adhere to the transformed epithelial cells and express murine mast cell protease 2 (mMCP2; a typical mucosal MC protease) and, to a lesser extent, the connective tissue mast cell (CTMC) protease mMCP6. Persistence of MCs is strictly dependent on T cell-derived IL-10, and their loss in the absence of IL-10-expressing T cells markedly delays small bowel (SB) polyposis. MCs expand profusely in polyposis-prone mice when T cells overexpress IL-10. The frequency of polyp-associated MCs is unaltered in response to broad-spectrum antibiotics, arguing against a microbial component driving their recruitment. Intriguingly, when polyps become invasive, a second wave of mMCP5+/mMCP6+ CTMCs expands in the tumor stroma and at invasive tumor borders. Ablation of mMCP6 expression attenuates polyposis, but invasive properties of the remaining lesions remain intact. Our findings argue for a multistep process in SB carcinogenesis in which distinct MC subsets, and their elaborated proteases, guide disease progression.
Asunto(s)
Quimasas/metabolismo , Citocinas/metabolismo , Neoplasias Intestinales/patología , Intestino Delgado/patología , Linfocitos/patología , Mastocitos/patología , Membrana Mucosa/patología , Animales , Células Cultivadas , Neoplasias Intestinales/inmunología , Neoplasias Intestinales/metabolismo , Intestino Delgado/inmunología , Intestino Delgado/metabolismo , Linfocitos/inmunología , Linfocitos/metabolismo , Mastocitos/inmunología , Mastocitos/metabolismo , Ratones , Membrana Mucosa/inmunología , Membrana Mucosa/metabolismo , Estadificación de NeoplasiasRESUMEN
Mast cells (MC) are potent innate immune cells that accumulate in chronically inflamed tissues. MC express the IL-33 receptor IL-1 receptor-related protein ST2 at high level, and this IL-1 family cytokine both activates MC directly and primes them to respond to other proinflammatory signals. Whether IL-33 and ST2 play a role in MC survival remains to be defined. In skin-derived human MC, we found that IL-33 attenuated MC apoptosis without altering proliferation, an effect mediated principally through the antiapoptotic molecule B-cell lymphoma-X large (BCLXL). Murine MC demonstrated a similar mechanism, dependent entirely on ST2. In line with these observations, St2(-/-) mice exhibited reduced numbers of tissue MC in inflamed arthritic joints, in helminth-infected intestine, and in normal peritoneum. To confirm an MC-intrinsic role for ST2 in vivo, we performed peritoneal transfer of WT and St2(-/-) MC. In St2(-/-) hosts treated with IL-33 and in WT hosts subjected to thioglycollate peritonitis, WT MC displayed a clear survival advantage over coengrafted St2(-/-) MC. IL-33 blockade specifically attenuated this survival advantage, confirming IL-33 as the relevant ST2 ligand mediating MC survival in vivo. Together, these data reveal a cell-intrinsic role for the IL-33/ST2 axis in the regulation of apoptosis in MC, identifying thereby a previously unappreciated pathway supporting expansion of the MC population with inflammation.
Asunto(s)
Interleucinas/metabolismo , Mastocitos/metabolismo , Receptores de Superficie Celular/metabolismo , Receptores de Interleucina/metabolismo , Proteína bcl-X/metabolismo , Animales , Artritis/genética , Artritis/inmunología , Artritis/metabolismo , Artritis/patología , Supervivencia Celular/genética , Supervivencia Celular/inmunología , Helmintiasis/genética , Helmintiasis/inmunología , Helmintiasis/metabolismo , Helmintiasis/patología , Humanos , Inflamación/inmunología , Inflamación/metabolismo , Inflamación/patología , Proteína 1 Similar al Receptor de Interleucina-1 , Interleucina-33 , Interleucinas/genética , Interleucinas/inmunología , Mucosa Intestinal/metabolismo , Intestinos/inmunología , Intestinos/parasitología , Articulaciones/inmunología , Articulaciones/metabolismo , Articulaciones/patología , Mastocitos/inmunología , Mastocitos/patología , Ratones , Ratones Noqueados , Receptores de Superficie Celular/genética , Receptores de Superficie Celular/inmunología , Receptores de Interleucina/genética , Receptores de Interleucina/inmunología , Proteína bcl-X/genética , Proteína bcl-X/inmunologíaRESUMEN
Papain, a cysteine protease allergen with inherent adjuvant activity, induces potent IL-4 expression by T cells in the popliteal lymph nodes of mice following footpad immunization. In this study, we identify a novel, non-BCR-mediated capacity for B cells to rapidly bind and internalize papain. B cells subsequently regulate the adaptive immune response by enhancing ICOS expression on CD4(+) T cells and amplifying Th2 and follicular helper T cell induction. Ab blockade of ICOS ligand, expressed by popliteal lymph node B cells, but not dendritic cells, at the peak of the response inhibits IL-4 responses in wild-type mice but not B cell-deficient mice. Thus, B cells play a critical role in amplifying adjuvant-dependent Th2 polarization following noncanonical acquisition and internalization of the cysteine protease papain.
Asunto(s)
Linfocitos B/inmunología , Linfocitos T CD4-Positivos/inmunología , Papaína/inmunología , Receptores de Antígenos de Linfocitos B/inmunología , Animales , Linfocitos B/metabolismo , Linfocitos T CD4-Positivos/metabolismo , Células Cultivadas , Citometría de Flujo , Inmunización/métodos , Proteína Coestimuladora de Linfocitos T Inducibles/inmunología , Proteína Coestimuladora de Linfocitos T Inducibles/metabolismo , Interleucina-4/inmunología , Interleucina-4/metabolismo , Ganglios Linfáticos/citología , Ganglios Linfáticos/inmunología , Ganglios Linfáticos/metabolismo , Ratones , Ratones de la Cepa 129 , Ratones Endogámicos C57BL , Ratones Noqueados , Ratones Transgénicos , Microscopía Confocal , Microscopía de Fluorescencia por Excitación Multifotónica , Papaína/administración & dosificación , Papaína/metabolismo , Unión Proteica/inmunología , Receptores de Antígenos de Linfocitos B/metabolismo , Células Th2/inmunología , Células Th2/metabolismoRESUMEN
Allergic asthma is a complex disease with a strong genetic component where mast cells play a major role by the release of proinflammatory mediators. In the mouse, mast cell protease-6 (mMCP-6) closely resembles the human version of mast cell tryptase, ß-tryptase. The gene that encodes mMCP-6, Tpsb2, resides close by the H-2 complex (MHC gene) on chromosome 17. Thus, when the original mMCP-6 knockout mice were backcrossed to the BALB/c strain, these mice were carrying the 129/Sv haplotype of MHC (mMCP-6(-/-)/H-2bc). Further backcrossing yielded mMCP-6(-/-) mice with the BALB/c MHC locus. BALB/c mice were compared with mMCP-6(-/-) and mMCP-6(-/-)/H-2bc mice in a mouse model of experimental asthma. Although OVA-sensitized and challenged wild type mice displayed a striking airway hyperresponsiveness (AHR), mMCP-6(-/-) mice had less AHR that was comparable with that of mMCP-6(-/-)/H-2bc mice, suggesting that mMCP-6 is required for a full-blown AHR. The mMCP-6(-/-)/H-2bc mice had strikingly reduced lung inflammation, IgE responses, and Th2 cell responses upon sensitization and challenge, whereas the mMCP-6(-/-) mice responded similarly to the wild type mice but with a minor decrease in bronchoalveolar lavage eosinophils. These findings suggest that inflammatory Th2 responses are highly dependent on the MHC-haplotype and that they can develop essentially independently of mMCP-6, whereas mMCP-6 plays a key role in the development of AHR.
Asunto(s)
Asma/inmunología , Hiperreactividad Bronquial/inmunología , Complejo Mayor de Histocompatibilidad , Mastocitos/inmunología , Células Th2/inmunología , Triptasas/inmunología , Animales , Asma/inducido químicamente , Asma/genética , Asma/patología , Hiperreactividad Bronquial/inducido químicamente , Hiperreactividad Bronquial/genética , Hiperreactividad Bronquial/patología , Líquido del Lavado Bronquioalveolar/química , Líquido del Lavado Bronquioalveolar/citología , Cromosomas de los Mamíferos , Cruzamientos Genéticos , Eosinófilos/inmunología , Eosinófilos/patología , Femenino , Regulación de la Expresión Génica , Antígenos H-2/genética , Antígenos H-2/inmunología , Haplotipos , Inmunoglobulina E/genética , Masculino , Mastocitos/patología , Ratones , Ratones Endogámicos BALB C , Ratones Noqueados , Ovalbúmina , Transducción de Señal , Células Th2/patología , Triptasas/genéticaRESUMEN
We previously established a mast cell (MC)-dependent thermal injury model in mice with ulceration and scar formation that depended on nonredundant functions of mouse MC protease (mMCP)4 and mMCP5. We hypothesized that MC activation is an early event and now find by histology that exocytosis of granule contents occurred by 2 min after thermal injury in wild-type (WT) C57BL/6 mice and in the mMCP4- or mMCP5-deficient mice. The degranulation was equivalent for MCs in the dermis and hypodermis of all three strains, but only the WT mice showed an appreciable increase in epidermal thickness. There was no loss of total MCs, partially degranulated plus intact, during the 4 h of observation. By electron microscopy, MCs in all strains showed early zonal degranulation at 30 s with marked progression in magnitude by 120 s and no mitochondrial injury or cellular necrosis. Concomitantly there was an increase in intercellular spaces indicative of tight junction (TJ) disruption in WT mice but not in the mMCP4- or mMCP5-deficient strains. The desmosomes were intact in all strains. Immunodetection of the TJ protein claudin 4 in WT and mMCP5-deficient mice indicated a significant reduction after scald injury whereas mMCP4(-/-) mice showed no significant changes. Taken together, these findings reveal that a second-degree burn injury can initiate an immediate novel zonal degranulation of MCs throughout all skin layers and a disruption of the epidermal TJs dependent on the nonredundant presence of mMCP4 and mMCP5.
Asunto(s)
Quimasas/deficiencia , Epidermis/metabolismo , Serina Endopeptidasas/deficiencia , Uniones Estrechas/metabolismo , Animales , Quemaduras/genética , Quemaduras/metabolismo , Degranulación de la Célula , Quimasas/genética , Claudina-4/metabolismo , Epidermis/lesiones , Epidermis/ultraestructura , Exocitosis , Técnica del Anticuerpo Fluorescente , Mastocitos/metabolismo , Mastocitos/fisiología , Mastocitos/ultraestructura , Ratones Endogámicos C57BL , Ratones Noqueados , Microscopía Electrónica , Serina Endopeptidasas/genética , Temperatura , Uniones Estrechas/patología , Uniones Estrechas/ultraestructura , Factores de TiempoRESUMEN
Mast cells are derived from committed progenitors that originate in the BM. They mature into histochemically distinguishable, metachromatic mast cells containing numerous cytoplasmic secretory granules. Accumulating evidence demonstrates that mast cell granule maturation is very tightly regulated by many factors including different granule components such as proteoglycans. In this issue of the European Journal of Immunology, Nakazawa et al. [Eur. J. Immunol. 2014. 44: 204-214] highlight a role for mast cell derived histamine as another factor critical for mast cell maturation. Using histidine decarboxylase (HDC) deficient mice that are unable to make histamine, they show poorly formed secretory granules and decreased secretory granule protease expression in peritoneal mast cells. Co-culturing BM-derived mast cells with fibroblasts normally drives granule maturation, but HDC-deficient BM-derived mast cells fail to do so. Exogenously provided histamine partly restores granule differentiation as evidenced by increased tryptase and chymase activity, and this is histamine receptor type H4 -dependent. However, H4 -deficient mice have intact granule formation in peritoneal mast cells, suggesting that when HDC is functional, the intrinsic histamine production is sufficient for most granule maturation processes and H4 is dispensable. This study highlights the role of histamine in the regulation of mast cell maturation, although the cytosolic target remains unknown.
Asunto(s)
Gránulos Citoplasmáticos/metabolismo , Fibroblastos/inmunología , Histamina/biosíntesis , Mastocitos/inmunología , Vesículas Secretoras/metabolismo , Animales , Femenino , MasculinoRESUMEN
Mast cells (MC) and basophils share expression of the high-affinity receptor for IgE (FcεRI) but can be distinguished by their divergent expression of KIT and CD49b. In BALB/c mice, MC lineage cells expressing high levels of FcεRI by flow cytometry were seen only in bone marrow whereas those expressing intermediate levels of FcεRI were present in bone marrow and spleen of naive mice and in mesenteric lymph nodes (mLN) of Trichinella spiralis-infected mice. These FcεRI(+)KIT(+)CD49b(-) cells had a membrane phenotype similar to i.p. connective tissue-type MC, but were smaller and hypogranular by flow cytometry forward and side scatter profiles, respectively. Consistent with this, they lacked the prominent secretory granules identified by histochemistry and immunodetection for the MC-specific granule proteases that are readily seen in mature jejunal mucosal MC that also are induced by the infection and present at the same time. The concentration of these MC lineage cells in mLN determined by flow cytometry was comparable to that of MC progenitors (MCp) measured by limiting dilution and clonal expansion with maturation. We observed upregulation of IL-4 transcription by MCp in mLN and spleens of helminth-infected 4get mice, and we demonstrated by intracellular cytokine staining production of IL-4 and IL-6 by the mLN MCp in helminth-infected mice. Furthermore, treatment of helminth-infected mice with anti-FcεRI mAb, a protocol known to deplete basophils, also depleted mLN MCp. Thus, this study identifies a hypogranular subset of MCp recruited to mLN by helminth infection that may be an important unrecognized source of cytokines.
Asunto(s)
Gránulos Citoplasmáticos/inmunología , Interleucina-4/biosíntesis , Interleucina-6/biosíntesis , Ganglios Linfáticos/inmunología , Mastocitos/inmunología , Triquinelosis/inmunología , Animales , Movimiento Celular/genética , Movimiento Celular/inmunología , Células Cultivadas , Gránulos Citoplasmáticos/parasitología , Gránulos Citoplasmáticos/patología , Regulación hacia Abajo/inmunología , Genes Reporteros , Interleucina-4/genética , Ganglios Linfáticos/parasitología , Ganglios Linfáticos/patología , Mastocitos/parasitología , Mastocitos/patología , Mesenterio/inmunología , Mesenterio/patología , Ratones , Ratones de la Cepa 129 , Ratones Endogámicos BALB C , Ratones Noqueados , Ratones Transgénicos , ARN Mensajero/biosíntesis , Células Madre/inmunología , Células Madre/parasitología , Células Madre/patología , Trichinella spiralis , Triquinelosis/parasitología , Triquinelosis/patología , Regulación hacia Arriba/genética , Regulación hacia Arriba/inmunologíaRESUMEN
Mouse mast cells (MCs) express a large number of serine proteases including tryptases, mouse mast cell protease (mMCP)-6 and -7; chymases, mMCP-1, -2, and -4; and an elastase, mMCP-5; along with carboxypeptidase-A3 (CPA3). In helminth-infected mouse intestine, distinct protease phenotypes are observed for connective tissue MCs (CTMCs) (mMCP-4(+)-7(+), and CPA3(+)) and mucosal MCs (MMCs) (mMCP-1(+) and 2(+)). To determine whether the protease phenotype was regulated by the tissue, we compared the phenotype of constitutive CTMCs and induced MMCs in trachea and large airways in antigen-sensitized unchallenged and challenged mice to MCs in skin and helminthic-infected intestine. We found that in the trachea, unlike in skin and intestine, CTMCs and MMCs both express all six serine proteases and CPA3 (mMCP-1(+), -2(+), 4(+)-7(+), CPA3(+)). This phenotype also holds for the lung CTMCs in the proximal bronchi, whereas the induced MMCs express only four proteases, mMCP-1, -2, -6, and -7. Thus, the T-cell-dependent induction of MMCs in trachea, large bronchi, and small intestine provides numbers but does not determine the protease phenotype. Furthermore, the CTMCs, which are constitutive, also show striking differences at these tissue sites, supporting the view that the differences in expression are tissue directed and not dependent on inflammation.
Asunto(s)
Células del Tejido Conectivo/enzimología , Mastocitos/enzimología , Membrana Mucosa/citología , Especificidad de Órganos , Péptido Hidrolasas/metabolismo , Aerosoles/metabolismo , Compuestos de Alumbre , Animales , Especificidad de Anticuerpos/inmunología , Antígenos/inmunología , Bronquios/citología , Recuento de Células , Diferenciación Celular , Células del Tejido Conectivo/citología , Inmunización , Mastocitos/citología , Ratones , Ratones Endogámicos BALB C , Ovalbúmina/inmunología , Fenotipo , Células Madre/citología , Factores de Tiempo , Tráquea/citologíaRESUMEN
BACKGROUND: Sensitization to food antigen can occur through cutaneous exposure. OBJECTIVE: We sought to test the hypothesis that epicutaneous sensitization with food antigen predisposes to IgE-mediated anaphylaxis on oral allergen challenge. METHODS: BALB/c mice were epicutaneously sensitized by repeated application of ovalbumin (OVA) to tape-stripped skin over 7 weeks or orally immunized with OVA and cholera toxin (CT) weekly for 8 weeks and then orally challenged with OVA. Body temperature was monitored, and serum mouse mast cell protease 1 levels were determined after challenge. Tissue mast cell (MC) counts were examined by using chloroacetate esterase staining. Levels of serum OVA-specific IgE and IgG(1) antibodies and cytokines in supernatants of OVA-stimulated splenocytes were measured by means of ELISA. Serum IL-4 levels were measured by using an in vivo cytokine capture assay. RESULTS: Epicutaneously sensitized mice exhibited expansion of connective tissue MCs in the jejunum, increased serum IL-4 levels, and systemic anaphylaxis after oral challenge, as evidenced by decreased body temperature and increased serum mouse mast cell protease 1 levels. Intestinal MC expansion and anaphylaxis were IgE dependent because they did not occur in epicutaneously sensitized IgE(-/-) mice. Mice orally immunized with OVA plus CT did not have increased serum IL-4 levels, expanded intestinal MCs, or anaphylaxis after oral challenge, despite OVA-specific IgE levels and splenocyte cytokine production in response to OVA stimulation, which were comparable with those of epicutaneously sensitized mice. CONCLUSION: Epicutaneously sensitized mice, but not mice orally immunized with antigen plus CT, have expansion of intestinal MCs and IgE-mediated anaphylaxis after single oral antigen challenge. IgE is necessary but not sufficient for food anaphylaxis, and MC expansion in the gut can play an important role in the development of anaphylaxis.
Asunto(s)
Anafilaxia/inmunología , Hipersensibilidad a los Alimentos/inmunología , Inmunoglobulina E/inmunología , Yeyuno/inmunología , Mastocitos/inmunología , Piel/inmunología , Administración Cutánea , Alérgenos/inmunología , Animales , Anticuerpos/inmunología , Antígenos/inmunología , Temperatura Corporal/inmunología , Quimiocina CCL2/inmunología , Toxina del Cólera/inmunología , Inmunoglobulina G/inmunología , Interleucina-4/inmunología , Ratones , Ratones Endogámicos BALB C , Ovalbúmina/inmunologíaRESUMEN
The transcription factor T-bet was identified in CD4(+) T cells, and it controls interferon gamma production and T helper type 1 cell differentiation. T-bet is expressed in certain other leukocytes, and we recently showed (Lord, G.M., R.M. Rao, H. Choe, B.M. Sullivan, A.H. Lichtman, F.W. Luscinskas, and L.H. Glimcher. 2005. Blood. 106:3432-3439) that it regulates T cell trafficking. We examined whether T-bet influences homing of mast cell progenitors (MCp) to peripheral tissues. Surprisingly, we found that MCp homing to the lung or small intestine in T-bet(-/-) mice is reduced. This is reproduced in adhesion studies using bone marrow-derived MCs (BMMCs) from T-bet(-/-) mice, which showed diminished adhesion to mucosal addresin cellular adhesion molecule-1 (MAdCAM-1) and vascular cell adhesion molecule-1 (VCAM-1), endothelial ligands required for MCp intestinal homing. MCp, their precursors, and BMMCs do not express T-bet, suggesting that T-bet plays an indirect role in homing. However, adoptive transfer experiments revealed that T-bet expression by BM cells is required for MCp homing to the intestine. Furthermore, transfer of WT BM-derived dendritic cells (DCs) to T-bet(-/-) mice restores normal MCp intestinal homing in vivo and MCp adhesion to MAdCAM-1 and VCAM-1 in vitro. Nonetheless, T-bet(-/-) mice respond vigorously to intestinal infection with Trichinella spiralis, eliminating a role for T-bet in MC recruitment to sites of infection and their activation and function. Therefore, remarkably, T-bet expression by DCs indirectly controls MCp homing to mucosal tissues.
Asunto(s)
Células de la Médula Ósea/metabolismo , Adhesión Celular/fisiología , Movimiento Celular/fisiología , Células Dendríticas/metabolismo , Mastocitos/citología , Células Madre/metabolismo , Proteínas de Dominio T Box/metabolismo , Animales , Western Blotting , Moléculas de Adhesión Celular/metabolismo , Cartilla de ADN , Mucosa Gástrica/metabolismo , Inmunohistoquímica , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Ratones Noqueados , Mucoproteínas , Mucosa Respiratoria/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Células Madre/fisiología , Proteínas de Dominio T Box/genética , Molécula 1 de Adhesión Celular Vascular/metabolismoRESUMEN
The serine protease chymase has been reported to generate intracardiac angiotensin-II (Ang-II) from Ang-I as well as an intermediate precursor of endothelin-1 (ET-1), ET-1 (1-31) from Big-ET-1. Although humans possess only one chymase, several murine isoforms are documented, each with its own specific catalytic activity. Among these, mouse mast cell protease 4 (mMCP-4) is the isoform most similar to the human chymase for its activity. The aim of this study was to characterize the capacity of mMCP-4 to convert Big-ET-1 into its bioactive metabolite, ET-1, in vitro and in vivo in the mouse model. Basal mean arterial pressure did not differ between wild-type (WT) and mMCP-4(-/-) mice. Systemic administration of Big-ET-1 triggered pressor responses and increased blood levels of immunoreactive (IR) ET-1 (1-31) and ET-1 that were reduced by more than 50% in mMCP-4 knockout (-/-) mice compared with WT controls. Residual responses to Big-ET-1 in mMCP-4(-/-) mice were insensitive to the enkephalinase/neutral endopeptidase inhibitor thiorphan and the specific chymase inhibitor TY-51469 {2-[4-(5-fluoro-3-methylbenzo[b]thiophen-2-yl)sulfonamido-3-methanesulfonylphenyl]thiazole-4-carboxylic acid}. Soluble fractions from the lungs, left cardiac ventricle, aorta, and kidneys of WT but not mMCP-4(-/-) mice generated ET-1 (1-31) from exogenous Big-ET-1 in a TY-51469-sensitive fashion as detected by high-performance liquid chromatography/ matrix-assisted laser desorption/ionization-mass spectrometry. Finally, pulmonary endogenous levels of IR-ET-1 were reduced by more than 40% in tissues derived from mMCP-4(-/-) mice compared with WT mice. Our results show that mMCP-4 plays a pivotal role in the dynamic conversion of systemic Big-ET-1 to ET-1 in the mouse model.
Asunto(s)
Aorta/enzimología , Endotelina-1/metabolismo , Ventrículos Cardíacos/enzimología , Serina Endopeptidasas/metabolismo , Animales , Aorta/efectos de los fármacos , Aorta/metabolismo , Carboxipeptidasas A/biosíntesis , Carboxipeptidasas A/genética , Carboxipeptidasas A/metabolismo , Resistencia a Medicamentos , Endotelina-1/análogos & derivados , Endotelina-1/sangre , Regulación Enzimológica de la Expresión Génica , Ventrículos Cardíacos/efectos de los fármacos , Ventrículos Cardíacos/metabolismo , Hemodinámica/efectos de los fármacos , Isoenzimas/biosíntesis , Isoenzimas/genética , Isoenzimas/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Especificidad de Órganos , Fragmentos de Péptidos/sangre , Fragmentos de Péptidos/metabolismo , Procesamiento Proteico-Postraduccional/efectos de los fármacos , Proteolisis/efectos de los fármacos , ARN Mensajero/metabolismo , Serina Endopeptidasas/química , Serina Endopeptidasas/genética , Inhibidores de Serina Proteinasa/farmacología , Sulfonamidas/farmacología , Tiofenos/farmacología , Tiorfan/farmacologíaRESUMEN
Nuclear factor of activated T cell (NFAT) transcription factors are key regulators of gene transcription within immune cells. The NFAT-interacting protein, (NIP45), augments NFAT-driven IL-4 expression by a mechanism that relies on arginine methylation. To establish the function of NIP45 in vivo, we generated mice with a targeted deletion of the gene encoding this cofactor. NIP45-deficient T helper cells displayed profound defects in the expression of NFAT-regulated cytokine genes, including IL-4. Whereas NIP45 deficiency does not interfere with T helper cell NFAT activation or lineage-specific transcription-factor expression, NIP45 acts as an enhancer for the assembly of protein arginine methyltransferase 1 and the protein arginine methyltransferase 1-linked histone 4 arginine 3 methylation with the IL-4 promoter. Our study reveals an essential role for NIP45 in promoting robust cytokine expression in vivo, which is required for the efficient handling of parasites. We propose that NIP45 acts as a molecular rheostat serving to amplify the type-2 immune response.
Asunto(s)
Citocinas/genética , Péptidos y Proteínas de Señalización Intracelular/metabolismo , Proteínas Nucleares/metabolismo , Células Th2/inmunología , Animales , Arginina/metabolismo , Ensamble y Desensamble de Cromatina , Eliminación de Gen , Regulación de la Expresión Génica , Histonas/metabolismo , Interleucina-4/genética , Péptidos y Proteínas de Señalización Intracelular/genética , Metilación , Ratones , Ratones Mutantes , Factores de Transcripción NFATC/metabolismo , Proteínas Nucleares/genética , Regiones Promotoras Genéticas , Proteína-Arginina N-Metiltransferasas/metabolismo , Trichinella spiralis , Triquinelosis/inmunologíaRESUMEN
In C57BL/6 mice, the recruitment of mast cell progenitors (MCps) to the lung is a feature of Ag-induced pulmonary inflammation that requires sensitization and challenge and is totally inhibited by the administration of anti-CD4 at the time of challenge. When mAb to TGFbeta1 or to IL-10R was administered at the time of challenge, the recruitment of MCp/10(6) mononuclear cells (MNCs) to the lung was inhibited by 56.3 and 69.6%, respectively, whereas mAb to IL-4, IFN-gamma, IL-6, IL-17A, and IL-17F had no effect. In sensitized and challenged C57BL/6 mice lacking TGFbetaRII on CD4(+) cells, the recruitment of MCp/10(6) MNCs was reduced by 67.8%. The requirement for TGFbeta1 and IL-10 suggested a role for CD4(+)CD25(+) T regulatory cells. Mice treated with anti-CD25 at the time of Ag-challenge showed a reduction in the recruitment of MCp/10(6) MNCs by 77.2% without any reduction in MNC influx. These results reveal an unexpected role for T regulatory cells in promoting the recruitment of MCps to the lungs of C57BL/6 mice with Ag-induced pulmonary inflammation.
Asunto(s)
Movimiento Celular/inmunología , Epítopos de Linfocito T/administración & dosificación , Pulmón/inmunología , Mastocitos/inmunología , Células Madre/inmunología , Linfocitos T Reguladores/inmunología , Animales , Anticuerpos Bloqueadores/administración & dosificación , Anticuerpos Monoclonales/administración & dosificación , Antígenos CD4/inmunología , Desensibilización Inmunológica , Epítopos de Linfocito T/inmunología , Inyecciones Intraperitoneales , Pulmón/citología , Masculino , Mastocitos/citología , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Ratones Noqueados , Ratones Transgénicos , Ovalbúmina/administración & dosificación , Ovalbúmina/inmunología , Células Madre/citología , Linfocitos T Reguladores/metabolismoRESUMEN
A second-degree epidermal scald burn in mice elicits an inflammatory response mediated by natural IgM directed to nonmuscle myosin with complement activation that results in ulceration and scarring. We find that such burn injury is associated with early mast cell (MC) degranulation and is absent in WBB6F1-Kit(W)/Kit(Wv) mice, which lack MCs in a context of other defects due to a mutation of the Kit receptor. To address further an MC role, we used transgenic strains with normal lineage development and a deficiency in a specific secretory granule component. Mouse strains lacking the MC-restricted chymase, mouse MC protease (mMCP)-4, or elastase, mMCP-5, show decreased injury after a second-degree scald burn, whereas mice lacking the MC-restricted tryptases, mMCP-6 and mMCP-7, or MC-specific carboxypeptidase A3 activity are not protected. Histologic sections showed some disruption of the epidermis at the scald site in the protected strains suggesting the possibility of topical reconstitution of full injury. Topical application of recombinant mMCP-5 or human neutrophil elastase to the scalded area increases epidermal injury with subsequent ulceration and scarring, both clinically and morphologically, in mMCP-5-deficient mice. Restoration of injury requires that topical administration of recombinant mMCP-5 occurs within the first hour postburn. Importantly, topical application of human MC chymase restores burn injury to scalded mMCP-4-deficient mice but not to mMCP-5-deficient mice revealing nonredundant actions for these two MC proteases in a model of innate inflammatory injury with remodeling.
Asunto(s)
Quemaduras/inmunología , Quimasas/inmunología , Cicatriz/inmunología , Epidermis/inmunología , Mastocitos/inmunología , Modelos Inmunológicos , Serina Endopeptidasas/inmunología , Animales , Quemaduras/enzimología , Quemaduras/genética , Quemaduras/patología , Carboxipeptidasas A/genética , Carboxipeptidasas A/inmunología , Carboxipeptidasas A/metabolismo , Degranulación de la Célula/genética , Degranulación de la Célula/inmunología , Quimasas/genética , Quimasas/metabolismo , Quimasas/farmacología , Cicatriz/enzimología , Cicatriz/genética , Cicatriz/patología , Epidermis/enzimología , Epidermis/patología , Humanos , Inmunoglobulina M/genética , Inmunoglobulina M/inmunología , Inmunoglobulina M/metabolismo , Inflamación , Elastasa de Leucocito/genética , Elastasa de Leucocito/inmunología , Elastasa de Leucocito/metabolismo , Elastasa de Leucocito/farmacología , Mastocitos/enzimología , Mastocitos/patología , Ratones , Ratones Endogámicos BALB C , Ratones Mutantes , Miosinas/genética , Miosinas/inmunología , Miosinas/metabolismo , Proteínas Proto-Oncogénicas c-kit , Serina Endopeptidasas/genética , Serina Endopeptidasas/metabolismo , Serina Endopeptidasas/farmacología , Triptasas/genética , Triptasas/inmunología , Triptasas/metabolismo , Triptasas/farmacologíaRESUMEN
Tissue-resident mast cells (MCs) are important in allergic diseases. In a mouse model of allergic airways inflammation, an increase in peribronchiolar MCs was associated with increased concentrations of the chemokine CCL2 in lung lavage. MC progenitors (MCps) arising in bone marrow (BM) are recruited to tissues by transendothelial migration, and we found that CCL2 is chemotactic for MCps in freshly isolated BM in vitro. Immature, but not mature, BM-derived MCs migrated in response to CCL2 when cultured in IL-3+stem cell factor (SCF) but not when cultured in IL-3 alone. However, the cells under both culture conditions expressed mRNA for CCR2, the receptor for CCL2, and bound the radiolabeled chemokine with similar affinities, highlighting SCF as a key mediator in coupling CCR2 to downstream events, culminating in chemotaxis. Immature BM-derived MCs from IL-3 +SCF cultures, when administered i.v., accumulated at skin sites injected with CCL2 in vivo. MCp recruitment to the allergen-sensitized/challenged lung was significantly reduced in CCR2(-/-) and CCL2(-/-) mouse strains. However, reconstitution studies of sublethally irradiated and BM-reconstituted mice indicated that BM cells and stromal elements could provide CCL2, whereas the CCR2 function resided with stromal elements rather than BM cells. These experiments revealed a new function of SCF in chemokine receptor coupling, but they suggest a complex role of the CCL2/CCR2 axis in recruiting MCps during pulmonary inflammation.