RESUMEN
Intestinal inflammation shifts microbiota composition and metabolism. How the host monitors and responds to such changes remains unclear. Here, we describe a protective mechanism by which mucosal-associated invariant T (MAIT) cells detect microbiota metabolites produced upon intestinal inflammation and promote tissue repair. At steady state, MAIT ligands derived from the riboflavin biosynthesis pathway were produced by aerotolerant bacteria residing in the colonic mucosa. Experimental colitis triggered luminal expansion of riboflavin-producing bacteria, leading to increased production of MAIT ligands. Modulation of intestinal oxygen levels suggested a role for oxygen in inducing MAIT ligand production. MAIT ligands produced in the colon rapidly crossed the intestinal barrier and activated MAIT cells, which expressed tissue-repair genes and produced barrier-promoting mediators during colitis. Mice lacking MAIT cells were more susceptible to colitis and colitis-driven colorectal cancer. Thus, MAIT cells are sensitive to a bacterial metabolic pathway indicative of intestinal inflammation.
Asunto(s)
Colitis , Disbiosis , Microbioma Gastrointestinal , Ratones Endogámicos C57BL , Células T Invariantes Asociadas a Mucosa , Animales , Células T Invariantes Asociadas a Mucosa/inmunología , Colitis/inmunología , Colitis/microbiología , Disbiosis/inmunología , Ratones , Microbioma Gastrointestinal/inmunología , Ratones Noqueados , Mucosa Intestinal/inmunología , Mucosa Intestinal/microbiología , Riboflavina/inmunologíaRESUMEN
The orf63 gene resides in a region of the lambda bacteriophage genome between the exo and xis genes and is among the earliest genes transcribed during infection. In lambda phage and Shiga toxin (Stx) producing phages found in enterohemorrhagic Escherichia coli (EHEC) associated with food poisoning, Orf63 expression reduces the host survival and hastens the period between infection and lysis thereby giving it pro-lytic qualities. The NMR structure of dimeric Orf63 reveals a fold consisting of two helices and one strand that all make extensive intermolecular contacts. Structure-based data mining failed to identify any Orf63 homolog beyond the family of temperate bacteriophages. A machine learning approach was used to design an amphipathic helical ligand that bound a hydrophobic cleft on Orf63 with micromolar affinity. This approach may open a new path towards designing therapeutics that antagonize the contributions of Stx phages in EHEC outbreaks.
Asunto(s)
Bacteriófago lambda , Escherichia coli Enterohemorrágica , Proteínas Virales , Bacteriófago lambda/genética , Bacteriófago lambda/metabolismo , Escherichia coli Enterohemorrágica/metabolismo , Escherichia coli Enterohemorrágica/virología , Toxina Shiga/genética , Proteínas Virales/metabolismoRESUMEN
In humans, MAIT cells represent the most abundant T cell subset reacting against bacteria. Their frequency in the blood is decreased in a large variety of infectious diseases of either bacterial or viral origin. MAIT cells accumulate at the site of bacterial infection and are protective in experimental infection models. Recent epidemiological evidence supports an implication of MAIT cells in protecting against tuberculosis. MAIT cells can be activated either through direct recognition of microbial ligands or by inflammatory cytokines such as IL-12 and IL-18. MAIT cells secrete IFN-γ, IL-17 and/or other effector molecules according to the context of triggering. MAIT cells can kill bacterially infected epithelial cells in vitro. Herein, we summarize and discuss the data suggesting a role for MAIT cells in infectious diseases.
Asunto(s)
Enfermedades Transmisibles/inmunología , Subgrupos de Linfocitos T/inmunología , Tuberculosis/inmunología , Animales , Apoptosis , Humanos , Interferón gamma/metabolismo , Interleucina-12/metabolismo , Interleucina-17/metabolismo , Interleucina-18/metabolismoRESUMEN
The MHC-related 1, MR1, molecule presents a new class of microbial antigens (derivatives of the riboflavin [Vitamin B2] biosynthesis pathway) to mucosal-associated invariant T (MAIT) cells. This raises many questions regarding antigens loading and intracellular trafficking of the MR1/ligand complexes. The MR1/MAIT field is also important because MAIT cells are very abundant in humans and their frequency is modified in many infectious and non-infectious diseases. Both MR1 and the invariant TCRα chain expressed by MAIT cells are strikingly conserved among species, indicating important functions. Riboflavin is synthesized by plants and most bacteria and yeasts but not animals, and its precursor derivatives activating MAIT cells are short-lived unless bound to MR1. The recognition of MR1 loaded with these compounds is therefore an exquisite manner to detect invasive bacteria. Herein, we provide an historical perspective of the field before describing the main characteristics of MR1, its ligands, and the few available data regarding its cellular biology. We then summarize the current knowledge of MAIT cell differentiation and discuss the definition of MAIT cells in comparison to related subsets. Finally, we describe the phenotype and effector activities of MAIT cells.
Asunto(s)
Antígenos de Histocompatibilidad Clase I/metabolismo , Inmunidad Mucosa , Antígenos de Histocompatibilidad Menor/metabolismo , Células T Invariantes Asociadas a Mucosa/inmunología , Riboflavina/inmunología , Subgrupos de Linfocitos T/inmunología , Animales , Diferenciación Celular , Antígenos de Histocompatibilidad/metabolismo , Humanos , Receptores de Antígenos de Linfocitos T alfa-beta/metabolismoRESUMEN
Mucosal-associated invariant T cells (MAITs) have potent antimicrobial activity and are abundant in humans (5%-10% in blood). Despite strong evolutionary conservation of the invariant TCR-α chain and restricting molecule MR1, this population is rare in laboratory mouse strains (≈0.1% in lymphoid organs), and lack of an appropriate mouse model has hampered the study of MAIT biology. Herein, we show that MAITs are 20 times more frequent in clean wild-derived inbred CAST/EiJ mice than in C57BL/6J mice. Increased MAIT frequency was linked to one CAST genetic trait that mapped to the TCR-α locus and led to higher usage of the distal Vα segments, including Vα19. We generated a MAIThi congenic strain that was then crossed to a transgenic Rorcgt-GFP reporter strain. Using this tool, we characterized polyclonal mouse MAITs as memory (CD44+) CD4-CD8lo/neg T cells with tissue-homing properties (CCR6+CCR7-). Similar to human MAITs, mouse MAITs expressed the cytokine receptors IL-7R, IL-18Rα, and IL-12Rß and the transcription factors promyelocytic leukemia zinc finger (PLZF) and RAR-related orphan receptor γ (RORγt). Mouse MAITs produced Th1/2/17 cytokines upon TCR stimulation and recognized a bacterial compound in an MR1-dependent manner. During experimental urinary tract infection, MAITs migrated to the bladder and decreased bacterial load. Our study demonstrates that the MAIThi congenic strain allows phenotypic and functional characterization of naturally occurring mouse MAITs in health and disease.
Asunto(s)
Ratones Congénicos/inmunología , Células T Asesinas Naturales/inmunología , Animales , Quimiotaxis de Leucocito , Cruzamientos Genéticos , Modelos Animales de Enfermedad , Femenino , Reordenamiento Génico de la Cadena alfa de los Receptores de Antígenos de los Linfocitos T , Vida Libre de Gérmenes , Antígenos de Histocompatibilidad Clase I/inmunología , Humanos , Memoria Inmunológica , Factores de Transcripción de Tipo Kruppel/análisis , Activación de Linfocitos , Recuento de Linfocitos , Tejido Linfoide/citología , Linfocinas/metabolismo , Ratones , Ratones Congénicos/genética , Ratones Congénicos/microbiología , Ratones Endogámicos C57BL , Ratones Noqueados , Ratones Transgénicos , Microbiota , Antígenos de Histocompatibilidad Menor , Células T Asesinas Naturales/metabolismo , Miembro 3 del Grupo F de la Subfamilia 1 de Receptores Nucleares/análisis , Fenotipo , Polimorfismo de Nucleótido Simple , Proteína de la Leucemia Promielocítica con Dedos de Zinc , Quimera por Radiación , Receptores de Antígenos de Linfocitos T alfa-beta/genética , Receptores de Citocinas/análisis , Infecciones Urinarias/inmunología , Infecciones Urinarias/microbiologíaRESUMEN
The lytic function of CTL relies on the polarized release of cytotoxic granules (CG) at the immune synapse (IS) with target cells. CTL also contain CCL5 in cytoplasmic storage vesicles (CCL5V) distinct from CG, the role of which, in regulating T cell effector functions, is not understood. Using human CD8(+) T cells specific to a lung tumor-associated Ag, we show in this article that CTL release both secretory compartments into the immune synapse with autologous tumor cells. Moreover, we demonstrate that disorganization of the T cell microtubule cytoskeleton and defects in hMunc13-4 or Rab27a abrogate CG exocytosis and synaptic secretion of the chemokine. Mechanistically, synaptic release of CCL5 cytoplasmic storage vesicles likely occurs upon their coalescence with the Rab27a-hMunc13-4 compartment and results in autocrine, CCR5-dependent induction of CXCR4 cell surface expression, thereby promoting T cell migration in response to CXCL12. We propose that CCL5 polarized delivery represents a mechanism by which CTL control immune synapse duration.
Asunto(s)
Antígenos de Neoplasias/inmunología , Quimiocina CCL5/inmunología , Quimiocina CXCL12/inmunología , Citotoxicidad Inmunológica , Receptores CXCR4/inmunología , Linfocitos T Citotóxicos/inmunología , Antígenos de Neoplasias/genética , Línea Celular Tumoral , Quimiocina CCL5/genética , Quimiocina CCL5/metabolismo , Quimiocina CXCL12/genética , Quimiotaxis , Gránulos Citoplasmáticos/química , Gránulos Citoplasmáticos/inmunología , Exocitosis/inmunología , Regulación de la Expresión Génica , Humanos , Sinapsis Inmunológicas , Microtúbulos/inmunología , Proteínas del Tejido Nervioso/deficiencia , Proteínas del Tejido Nervioso/genética , Proteínas del Tejido Nervioso/inmunología , Receptores CXCR4/genética , Transducción de Señal , Linfocitos T Citotóxicos/patología , Proteínas de Unión al GTP rab/deficiencia , Proteínas de Unión al GTP rab/genética , Proteínas de Unión al GTP rab/inmunología , Proteínas rab27 de Unión a GTPRESUMEN
T-cell adhesion/costimulatory molecules and their cognate receptors on target cells play a major role in T-cell receptor (TCR)-mediated activities. Here, we compared the involvement of CD103 and LFA-1, and their respective ligands, in the maturation of the cytotoxic immune synapse (cIS) and in the activation of CTL effector functions. Our results indicate that cytotoxicity toward cancer cells and, to a lesser extent, cytokine production by specific CTL require, together with TCR engagement, the interaction of either CD103 with E-cadherin or LFA-1 with ICAM-1. Flow-based adhesion assay showed that engagement of CD103 or LFA-1, together with TCR, enhances the strength of the T-cell/target cell interaction. Moreover, electron microscopic analyses showed that integrin-dependent mature cIS (mcIS) displays a cohesive ultrastructure, with tight membrane contacts separated by extensive clefts. In contrast, immature cIS (icIS), which is unable to trigger target cell lysis, is loose, with multiple protrusions in the effector cell membrane. Experiments using confocal microscopy revealed polarized cytokine release and degranulation at the mcIS associated with target cell killing, whereas icIS is characterized by failure of IFN-γ and granzyme B relocalization. Thus, interactive forces between CTL and epithelial tumor cells, mainly regulated by integrin engagement, correlate with maturity and the ultrastructure of the cIS and influence CTL effector functions. These results provide new insights into molecular mechanisms regulating antitumor CTL responses and may lead to the development of more efficient cancer immunotherapy strategies.
Asunto(s)
Antígenos CD/metabolismo , Citotoxicidad Inmunológica , Cadenas alfa de Integrinas/metabolismo , Neoplasias Pulmonares/inmunología , Antígeno-1 Asociado a Función de Linfocito/metabolismo , Linfocitos T Citotóxicos/inmunología , Cadherinas/metabolismo , Línea Celular Tumoral , Humanos , Molécula 1 de Adhesión Intercelular/metabolismo , Receptores de Antígenos de Linfocitos T/inmunologíaRESUMEN
Immune system-mediated eradication of neoplastic cells requires induction of a strong long-lasting antitumor T-cell response. However, generation of tumor-specific effector T cells does not necessarily result in tumor clearance. CTL must first be able to migrate to the tumor site, infiltrate the tumor tissue, and interact with the target to finally trigger effector functions indispensable for tumor destruction. Chemokines are involved in circulation, homing, retention, and activation of immunocompetent cells. Although some of them are known to contribute to tumor growth and metastasis, others are responsible for changes in the tumor microenvironment that lead to extensive infiltration of lymphocytes, resulting in tumor eradication. Given their chemoattractive and activating properties, a role for chemokines in the development of the effector phase of the antitumor immune response has been suggested. Here, we emphasize the role of the chemokine-chemokine receptor network at multiple levels of the T-cell-mediated antitumor immune response. The identification of chemokine-dependent molecular mechanisms implicated in tumor-specific CTL trafficking, retention, and regulation of their in situ effector functions may offer new perspectives for development of innovative immunotherapeutic approaches to cancer treatment.
Asunto(s)
Quimiocinas/fisiología , Inmunidad Celular , Neoplasias/inmunología , Receptores de Quimiocina/fisiología , Animales , Quimiocinas/genética , Quimiocinas/metabolismo , Quimiotaxis de Leucocito/genética , Quimiotaxis de Leucocito/inmunología , Citotoxicidad Inmunológica/genética , Citotoxicidad Inmunológica/fisiología , Humanos , Inmunidad Celular/genética , Inmunidad Celular/inmunología , Inmunidad Celular/fisiología , Modelos Biológicos , Receptores de Quimiocina/genética , Receptores de Quimiocina/metabolismo , Linfocitos T Citotóxicos/metabolismo , Linfocitos T Citotóxicos/fisiologíaRESUMEN
Appropriate localization and migration of T cells is a prerequisite for antitumor immune surveillance. Studies using fixed tumor samples from human patients have shown that T cells accumulate more efficiently in the stroma than in tumor islets, but the mechanisms by which this occurs are unknown. By combining immunostaining and real-time imaging in viable slices of human lung tumors, we revealed that the density and the orientation of the stromal extracellular matrix likely play key roles in controlling the migration of T cells. Active T cell motility, dependent on chemokines but not on ß1 or ß2 integrins, was observed in loose fibronectin and collagen regions, whereas T cells migrated poorly in dense matrix areas. Aligned fibers in perivascular regions and around tumor epithelial cell regions dictated the migratory trajectory of T cells and restricted them from entering tumor islets. Consistently, matrix reduction with collagenase increased the ability of T cells to contact cancer cells. Thus, the stromal extracellular matrix influences antitumor immunity by controlling the positioning and migration of T cells. Understanding the mechanisms by which this collagen network is generated has the potential to aid in the development of new therapeutics.
Asunto(s)
Neoplasias Pulmonares/patología , Linfocitos T/citología , Animales , Vasos Sanguíneos/patología , Antígenos CD18/metabolismo , Movimiento Celular , Colagenasas/metabolismo , Matriz Extracelular/metabolismo , Humanos , Sistema Inmunológico , Integrina beta1/metabolismo , Ligandos , Pulmón/metabolismo , Ratones , Ratones SCID , Modelos Biológicos , Trasplante de NeoplasiasRESUMEN
The CD5 coreceptor is expressed on all T cells and on the B1a B cell subset. It is associated with TCR and BCR, and modulates intracellular signals initiated by both Ag receptor complexes. Human CD5 contributes to regulation of the antitumor immune response and susceptibility of specific CTL to activation-induced cell death (AICD) triggered by the tumor. In this study, we compared the T cell response to the B16F10 melanoma engrafted into CD5-deficient and wild-type C57BL/6 mice. Compared with wild-type mice, CD5 knockout animals displayed delayed tumor growth, associated with tumor infiltration by T cell populations exhibiting a more activated phenotype and enhanced antitumor effector functions. However, control of tumor progression in CD5(-/-) mice was transient due to increased AICD of CD8(+) tumor-infiltrating T lymphocytes. Remarkably, in vivo protection of T cells from TCR-mediated apoptosis by an adenovirus engineered to produce soluble Fas resulted in a dramatic reduction in tumor growth. Our data suggest that recruitment of tumor-specific T cells in the tumor microenvironment occurs at early stages of cancer development and that tumor-mediated AICD of tumor-infiltrating T lymphocytes is most likely involved in tumor escape from the immune system.
Asunto(s)
Antígenos CD5/genética , Activación de Linfocitos/inmunología , Linfocitos Infiltrantes de Tumor/inmunología , Linfocitos Infiltrantes de Tumor/patología , Melanoma Experimental/prevención & control , Linfocitos T Citotóxicos/inmunología , Linfocitos T Citotóxicos/patología , Regulación hacia Arriba/inmunología , Animales , Antígenos CD5/metabolismo , Carcinoma Pulmonar de Lewis/genética , Carcinoma Pulmonar de Lewis/patología , Carcinoma Pulmonar de Lewis/prevención & control , Muerte Celular/genética , Muerte Celular/inmunología , Línea Celular Tumoral , Citotoxicidad Inmunológica/genética , Humanos , Tolerancia Inmunológica/genética , Activación de Linfocitos/genética , Melanoma Experimental/genética , Melanoma Experimental/patología , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Receptores de Antígenos de Linfocitos T/biosíntesis , Receptores de Antígenos de Linfocitos T/genética , Linfocitos T Citotóxicos/metabolismo , Regulación hacia Arriba/genéticaRESUMEN
Interaction of the integrin αE(CD103)ß7 expressed on tumor-infiltrating lymphocytes (TIL) with E-cadherin on epithelial tumor cells is required to trigger polarized exocytosis of cytotoxic granules in TIL that elicit tumor cell lysis. In this study, we investigated the functional and signaling properties of CD103 and its individual contribution to T-cell-mediated cancer-cell killing. Our results indicated that the binding of CD103 on tumor-specific CTL to immobilized recombinant E-cadherin-Fc is sufficient to induce the polarization of cytolytic granules, whereas the degranulation of cytolytic granules also requires the coengagement of the T-cell receptor. Moreover, minimal CD103 triggering promotes the phosphorylation of the ERK1/2 kinases and phospholipase Cγ1 (PLCγ1). Inhibiting PLCγ blocks granule relocalization, decreasing T-cell receptor-mediated cytotoxicity. Thus, our results emphasize a unique costimulatory role of CD103 in tumor-specific CTL activation by providing signals that promote T-cell effector functions needed to specifically target and lyse cancer cells.
Asunto(s)
Antígenos CD/inmunología , Cadherinas/inmunología , Fragmentos Fc de Inmunoglobulinas/inmunología , Cadenas alfa de Integrinas/inmunología , Neoplasias Pulmonares/inmunología , Fosfolipasa C gamma/inmunología , Linfocitos T Citotóxicos/inmunología , Antígenos CD/metabolismo , Cadherinas/química , Adhesión Celular/inmunología , Línea Celular Tumoral , Gránulos Citoplasmáticos/inmunología , Gránulos Citoplasmáticos/metabolismo , Citotoxicidad Inmunológica , Humanos , Proteínas Inmovilizadas/química , Proteínas Inmovilizadas/inmunología , Fragmentos Fc de Inmunoglobulinas/química , Cadenas alfa de Integrinas/metabolismo , Neoplasias Pulmonares/metabolismo , Proteína Quinasa 1 Activada por Mitógenos/inmunología , Proteína Quinasa 1 Activada por Mitógenos/metabolismo , Proteína Quinasa 3 Activada por Mitógenos/inmunología , Proteína Quinasa 3 Activada por Mitógenos/metabolismo , Fosfatidilinositol 3-Quinasas/inmunología , Fosfatidilinositol 3-Quinasas/metabolismo , Fosfolipasa C gamma/antagonistas & inhibidores , Fosfolipasa C gamma/metabolismo , Fosforilación , Proteína Quinasa C/inmunología , Proteína Quinasa C/metabolismo , Proteínas Recombinantes de Fusión/química , Proteínas Recombinantes de Fusión/inmunología , Linfocitos T Citotóxicos/metabolismoRESUMEN
We have reported previously that the interaction of alpha(E)(CD103)beta(7) integrin, expressed on a CD8(+) tumor-infiltrating lymphocyte (TIL) clone but not on a peripheral blood lymphocyte (PBL) counterpart, with the epithelial marker E-cadherin on human lung tumor cells plays a crucial role in T-cell receptor-mediated cytotoxicity. We show here that both TIL and PBL clones are able to migrate toward autologous tumor cells and that chemokine receptor CCR5 is involved in this process. Adoptive transfer of the PBL clone in the cognate tumor engrafted in nonobese diabetic/severe combined immunodeficient mice and subsequent coengagement of T-cell receptor and transforming growth factor-beta1 receptor triggers CD103 expression on T-cell surface resulting in strong potentiation of antitumor lytic function. Moreover, interaction of alpha(E)beta(7) integrin with E-cadherin, but not lymphocyte function-associated antigen-1 with intercellular adhesion molecule-1, promotes CCR5 recruitment at the immunologic synapse formed between TIL and tumor cells, leading to inhibition of T-cell sensitivity to CCL5 chemotactic gradient. These results provide evidence for a role of tumor microenvironment, namely MHC class I-restricted antigen presentation and transforming growth factor-beta1 secretion, in regulating the effector phase of tumor-specific CTL response. They also suggest a unique role of CD103 in T-cell retention at the tumor site by a CCR5-dependent mechanism.
Asunto(s)
Antígenos CD/biosíntesis , Carcinoma de Pulmón de Células no Pequeñas/inmunología , Cadenas alfa de Integrinas/biosíntesis , Neoplasias Pulmonares/inmunología , Receptores CCR5/inmunología , Linfocitos T Citotóxicos/inmunología , Animales , Antígenos CD/inmunología , Cadherinas/inmunología , Carcinoma de Pulmón de Células no Pequeñas/sangre , Línea Celular Tumoral , Movimiento Celular/inmunología , Humanos , Cadenas alfa de Integrinas/inmunología , Molécula 1 de Adhesión Intercelular/inmunología , Neoplasias Pulmonares/sangre , Antígeno-1 Asociado a Función de Linfocito/inmunología , Linfocitos/inmunología , Linfocitos Infiltrantes de Tumor/inmunología , Ratones , Ratones Endogámicos NOD , Ratones Desnudos , Ratones SCID , Receptores de Antígenos de Linfocitos T/biosíntesis , Receptores de Antígenos de Linfocitos T/inmunología , Receptores CCR5/biosíntesisRESUMEN
Chemokine receptors (CCR2 and CXCR4) are used as coreceptors for entry of human immunodeficiency virus (HIV) into the target cells. Mutations in CCR2 (CCR2-64I) and stromal-derived factor SDF1 (SDF1-3'A), the primary ligand for CXCR4, exhibited a protective effect against the onset of acquired immune deficiency syndrome (AIDS). The frequency of the SDF1-3'A and CCR2-64I alleles were determined in blood donors from 16 provinces, covering the entire territory of Poland. Of 1063 individuals, 274 (25.8%) were carriers of the SDF1-3'A allele; 36 of them (3.4%) were homozygotes (SDF-3'A/A) while 238 (22.4%) were heterozygotes (SDF-3'G/A), resulting in a 14.6% frequency of the SDF1-3'A allele. Moreover, in the same group of individuals, 234 (22.0%) carried the CCR2-64I allele; 6 of them (0.6%) were homozygotes (CCR2-64I/I), and 228 (21.4%) were heterozygotes (CCR2-64V/I), resulting in an 11.3% frequency of the CCR2-64I allele. The highest frequencies of the SDF1-3'A allele were found in the northeastern provinces and in one of the western provinces of Poland. In contrast, allelic frequencies of CCR2-64I varied slightly among different provinces. The different pattern of prevalence of the SDF1-3'A and CCR2-64I alleles in Poland might suggest that the CCR2-64I allele was spread much earlier than the SDF1-3'A allele in the population of Poland.