RESUMEN
Breach of tolerance to gluten leads to the chronic small intestinal enteropathy celiac disease. A key event in celiac disease development is gluten-dependent infiltration of activated cytotoxic intraepithelial lymphocytes (IELs), which cytolyze epithelial cells causing crypt hyperplasia and villous atrophy. The mechanisms leading to gluten-dependent small intestinal IEL infiltration and activation remain elusive. We have demonstrated that under homeostatic conditions in mice, gluten drives the differentiation of anti-inflammatory T cells producing large amounts of the immunosuppressive cytokine interleukin-10 (IL-10). Here we addressed whether this dominant IL-10 axis prevents gluten-dependent infiltration of activated cytotoxic IEL and subsequent small intestinal enteropathy. We demonstrate that IL-10 regulation prevents gluten-induced cytotoxic inflammatory IEL infiltration. In particular, IL-10 suppresses gluten-induced accumulation of a specialized population of cytotoxic CD4+CD8αα+ IEL (CD4+ CTL) expressing Tbx21, Ifng, and Il21, and a disparate non-cytolytic CD4+CD8α- IEL population expressing Il17a, Il21, and Il10. Concomitantly, IL-10 suppresses gluten-dependent small intestinal epithelial hyperproliferation and upregulation of stress-induced molecules on epithelial cells. Remarkably, frequencies of granzyme B+CD4+CD8α+ IEL are increased in pediatric celiac disease patient biopsies. These findings demonstrate that IL-10 is pivotal to prevent gluten-induced small intestinal inflammation and epithelial damage, and imply that CD4+ CTL are potential new players into these processes.
Asunto(s)
Linfocitos T CD4-Positivos/inmunología , Enfermedad Celíaca/inmunología , Interleucina-10/metabolismo , Mucosa Intestinal/inmunología , Linfocitos Intraepiteliales/inmunología , Animales , Muerte Celular , Diferenciación Celular , Movimiento Celular , Niño , Citotoxicidad Inmunológica , Glútenes/inmunología , Granzimas/metabolismo , Homeostasis , Humanos , Tolerancia Inmunológica , Interleucina-10/genética , Activación de Linfocitos , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Transducción de Señal , Proteínas de Dominio T Box/genética , Proteínas de Dominio T Box/metabolismoRESUMEN
Tolerance to harmless exogenous antigens is the default immune response in the gastrointestinal tract. Although extensive studies have demonstrated the importance of the mesenteric lymph nodes (MLNs) and intestinal CD103(+) dendritic cells (DCs) in driving small intestinal tolerance to protein antigen, the structural and immunological basis of colonic tolerance remain poorly understood. We show here that the caudal and iliac lymph nodes (ILNs) are inductive sites for distal colonic immune responses and that colonic T cell-mediated tolerance induction to protein antigen is initiated in these draining lymph nodes and not in MLNs. In agreement, colonic tolerance induction was not altered by mesenteric lymphadenectomy. Despite tolerance development, CD103(+)CD11b(+) DCs, which are the major migratory DC population in the MLNs, and the tolerance-related retinoic acid-generating enzyme RALDH2 were virtually absent from the ILNs. Administration of ovalbumin (OVA) to the distal colon did increase the number of CD11c(+)MHCII(hi) migratory CD103(-)CD11b(+) and CD103(+)CD11b(-) DCs in the ILNs. Strikingly, colonic tolerance was intact in Batf3-deficient mice specifically lacking CD103(+)CD11b(-) DCs, suggesting that CD103(-) DCs in the ILNs are sufficient to drive tolerance induction after protein antigen encounter in the distal colon. Altogether, we identify different inductive sites for small intestinal and colonic T-cell responses and reveal that distinct cellular mechanisms are operative to maintain tolerance at these sites.
Asunto(s)
Colon/inmunología , Células Dendríticas/inmunología , Intestino Delgado/inmunología , Ganglios Linfáticos/inmunología , Linfocitos T/inmunología , Aldehído Oxidorreductasas/genética , Aldehído Oxidorreductasas/metabolismo , Animales , Antígenos CD/metabolismo , Factores de Transcripción con Cremalleras de Leucina de Carácter Básico/genética , Antígeno CD11b/metabolismo , Femenino , Vena Ilíaca/anatomía & histología , Tolerancia Inmunológica , Cadenas alfa de Integrinas/metabolismo , Escisión del Ganglio Linfático , Ganglios Linfáticos/anatomía & histología , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Ratones Noqueados , Ratones Transgénicos , Proteínas Represoras/genéticaRESUMEN
Intestinal lymphoid tissues have to simultaneously ensure protection against pathogens and tolerance toward commensals. Despite such vital functions, their development in the colon is poorly understood. Here, we show that the two distinct lymphoid tissues of the colon-colonic patches and colonic solitary intestinal lymphoid tissues (SILTs)-can easily be distinguished based on anatomical location, developmental timeframe, and cellular organization. Furthermore, whereas colonic patch development depended on CXCL13-mediated lymphoid tissue inducer (LTi) cell clustering followed by LTα-mediated consolidation, early LTi clustering at SILT anlagen did not require CXCL13, CCR6, or CXCR3. Subsequent dendritic cell recruitment to and gp38(+)VCAM-1(+) lymphoid stromal cell differentiation within SILTs required LTα; B-cell recruitment and follicular dendritic cell differentiation depended on MyD88-mediated signaling, but not the microflora. In conclusion, our data demonstrate that different mechanisms, mediated mainly by programmed stimuli, induce the formation of distinct colonic lymphoid tissues, therefore suggesting that these tissues may have different functions.
Asunto(s)
Linfocitos B/inmunología , Colon/inmunología , Células Dendríticas/inmunología , Tejido Linfoide/inmunología , Linfotoxina-alfa/metabolismo , Células del Estroma/inmunología , Animales , Diferenciación Celular , Movimiento Celular , Células Cultivadas , Quimiocina CXCL13/genética , Quimiocina CXCL13/metabolismo , Colon/anatomía & histología , Tejido Linfoide/citología , Tejido Linfoide/crecimiento & desarrollo , Linfotoxina-alfa/inmunología , Glicoproteínas de Membrana/metabolismo , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Factor 88 de Diferenciación Mieloide/metabolismo , Receptores CCR6/genética , Receptores CCR6/metabolismo , Receptores CXCR3/metabolismo , Transducción de Señal , Molécula 1 de Adhesión Celular Vascular/metabolismoRESUMEN
We identified an IL-7Ralpha(+)Sca-1(low)c-Kit(low) population in E14 fetal liver, which is the phenotypical analog of common lymphoid progenitors (CLP) in adult bone marrow. After transfer into newborn mice, the IL-7Ralpha(+)Sca-1(low)c-Kit(low) population rapidly differentiated into CD45(+)CD4(+)CD3(-) cells, which are candidate cells for initiating lymph node and Peyer's patch formation. In addition, this population also gave rise to B, T, NK, and CD8alpha(+) and CD8alpha(-) dendritic cells. The fetal liver precursors expressed a significantly lower level of the myeloid-suppressing transcription factor Pax-5, than adult CLP, and retained differentiation activity for macrophages in vitro. We propose that the transition from fetal liver IL-7Ralpha(+)Sca-1(low)c-Kit(low) cells to adult CLP involves a regulated restriction of their developmental potential, controlled, at least in part, by Pax-5 expression.
Asunto(s)
Complejo CD3/biosíntesis , Antígenos CD4/biosíntesis , Antígenos Comunes de Leucocito/biosíntesis , Hígado/embriología , Hígado/inmunología , Subgrupos Linfocitarios/citología , Macrófagos/citología , Células Madre/inmunología , Animales , Animales Recién Nacidos/inmunología , Antígenos Ly/biosíntesis , Linfocitos B/citología , Diferenciación Celular/inmunología , Linaje de la Célula/inmunología , Células Cultivadas , Células Dendríticas/trasplante , Células Precursoras Eritroides/citología , Feto/citología , Feto/inmunología , Regulación del Desarrollo de la Expresión Génica/inmunología , Inmunofenotipificación , Hígado/citología , Hígado/metabolismo , Trasplante de Hígado/inmunología , Ganglios Linfáticos/citología , Subgrupos Linfocitarios/metabolismo , Subgrupos Linfocitarios/trasplante , Macrófagos/metabolismo , Proteínas de la Membrana/biosíntesis , Ratones , Ratones Endogámicos C57BL , Células Mieloides/citología , Proteínas Proto-Oncogénicas c-kit/biosíntesis , Receptores de Interleucina-7/biosíntesis , Trasplante de Células Madre , Células Madre/citología , Células Madre/metabolismoRESUMEN
Proper lymph node (LN) development requires tumor necrosis factor-related activation-induced cytokine (TRANCE) expression. Here we demonstrate that the defective LN development in TRANCE(-/)- mice correlates with a significant reduction in lymphotoxin (LT)alphabeta(+)alpha(4)beta(7)(+)CD45(+)CD4(+)CD3(-) cells and their failure to form clusters in rudimentary mesenteric LNs. Transgenic TRANCE overexpression in TRANCE(-/)- mice results in selective restoration of this cell population into clusters, and results in full LN development. Transgenic TRANCE-mediated restoration of LN development requires LTalphabeta expression on CD45(+) CD4(+)CD3(-) cells, as LNs could not be induced in LTalpha(-/)- mice. LTalpha(-/)- mice also showed defects in the fate of CD45(+)CD4(+)CD3(-) cells similar to TRANCE(-/)- mice. Thus, we propose that both TRANCE and LTalphabeta regulate the colonization and cluster formation by CD45(+) CD4(+)CD3(-) cells in developing LNs, the degree of which appears to correlate with the state of LN organogenesis.