RESUMEN
The study of human macrophages and their ontogeny is an important unresolved issue. Here, we use a humanized mouse model expressing human cytokines to dissect the development of lung macrophages from human hematopoiesis in vivo. Human CD34+ hematopoietic stem and progenitor cells (HSPCs) generated three macrophage populations, occupying separate anatomical niches in the lung. Intravascular cell labeling, cell transplantation, and fate-mapping studies established that classical CD14+ blood monocytes derived from HSPCs migrated into lung tissue and gave rise to human interstitial and alveolar macrophages. In contrast, non-classical CD16+ blood monocytes preferentially generated macrophages resident in the lung vasculature (pulmonary intravascular macrophages). Finally, single-cell RNA sequencing defined intermediate differentiation stages in human lung macrophage development from blood monocytes. This study identifies distinct developmental pathways from circulating monocytes to lung macrophages and reveals how cellular origin contributes to human macrophage identity, diversity, and localization in vivo.
Asunto(s)
Células Madre Hematopoyéticas/inmunología , Pulmón/inmunología , Macrófagos Alveolares/inmunología , Monocitos/inmunología , Antígenos CD34/metabolismo , Biodiversidad , Diferenciación Celular , Movimiento Celular , Células Cultivadas , Sangre Fetal/citología , Humanos , Receptores de Lipopolisacáridos/metabolismo , Pulmón/irrigación sanguínea , Receptores de IgG/metabolismo , Análisis de Secuencia de ARN , Análisis de la Célula Individual , Nicho de Células MadreRESUMEN
The innate responsiveness of the immune system is important not only for quick responses to pathogens but also for the initiation and shaping of the subsequent adaptive immune response. Activation via the cytokine IL-18, a product of inflammasomes, gives rise to a rapid response that includes the production of self-reactive antibodies. As increased concentrations of this cytokine are found in inflammatory diseases, we investigated the origin of the B cell response and its regulation. We identified an accumulation of B cell-helper neutrophils in the spleen that interacted with innate-type invariant natural killer T cells (iNKT cells) to regulate B cell responses. We found that neutrophil-dependent expression of the death-receptor ligand FasL by iNKT cells was needed to restrict autoantibody production. Neutrophils can thus license iNKT cells to regulate potentially harmful autoreactive B cell responses during inflammasome-driven inflammation.
Asunto(s)
Linfocitos B/inmunología , Proteína Ligando Fas/metabolismo , Inflamación/inmunología , Células T Asesinas Naturales/inmunología , Neutrófilos/inmunología , Inmunidad Adaptativa , Animales , Autoanticuerpos/biosíntesis , Células Cultivadas , Proteína Ligando Fas/genética , Inmunidad Innata , Inflamasomas/metabolismo , Interleucina-18/metabolismo , Ratones , Ratones Endogámicos C57BL , Ratones NoqueadosRESUMEN
Invariant natural killer T (iNKT) cells mediate rapid immune responses which bridge the gap between innate and adaptive responses to pathogens while also providing key regulation to maintain immune homeostasis. Both types of important iNKT immune responses are mediated through interactions with innate and adaptive B cells. As such, iNKT cells sit at the decision-making fulcrum between regulating inflammatory or autoreactive B cells and supporting protective or regulatory B cell populations. iNKT cells interpret the signals in their environment to set the tone for subsequent adaptive responses, with outcomes ranging from getting licensed to maintain homeostasis as an iNKT regulatory cell (iNKTreg ) or being activated to become an iNKT follicular helper (iNKTFH ) cell supporting pathogen-specific effector B cells. Here we review iNKT and B cell cooperation across the spectrum of immune outcomes, including during allergy and autoimmune disease, tumor surveillance and immunotherapy, or pathogen defense and vaccine responses. Because of their key role as influencers, iNKT cells provide a valuable target for therapeutic interventions. Understanding the nature of the interactions between iNKT and B cells will enable the development of clinical interventions to strategically target regulatory iNKT and B cell populations or inflammatory ones, depending on the circumstance.
Asunto(s)
Linfocitos B Reguladores , Células T Asesinas Naturales , Inmunidad Innata , Inmunoterapia , Activación de LinfocitosRESUMEN
The transcription factor Blimp1 is not only an essential regulator of plasma cells, but also a risk factor for the development of autoimmune disease in humans. Here, we demonstrate in the mouse that the Prdm1 (Blimp1) gene was partially activated at the chromatin and transcription level in early B cell development, although mature Prdm1 mRNA did not accumulate due to posttranscriptional regulation. By analyzing a mouse model that facilitated ectopic Blimp1 protein expression throughout B lymphopoiesis, we could demonstrate that Blimp1 impaired B cell development by interfering with the B cell gene expression program, while leading to an increased abundance of plasma cells by promoting premature plasmablast differentiation of immature and mature B cells. With progressing age, these mice developed an autoimmune disease characterized by the presence of autoantibodies and glomerulonephritis. Hence, these data identified ectopic Blimp1 expression as a novel mechanism, through which Blimp1 can act as a risk factor in the development of autoimmune disease.
Asunto(s)
Linfocitos B/metabolismo , Glomerulonefritis/metabolismo , Factor 1 de Unión al Dominio 1 de Regulación Positiva/genética , Factor 1 de Unión al Dominio 1 de Regulación Positiva/metabolismo , Animales , Autoanticuerpos/metabolismo , Linfocitos B/citología , Diferenciación Celular , Modelos Animales de Enfermedad , Femenino , Redes Reguladoras de Genes , Glomerulonefritis/genética , Humanos , Masculino , Ratones , Activación TranscripcionalRESUMEN
Invariant natural killer T (iNKT) cells serve as early rapid responders in the innate immune response to self-derived autoantigens and pathogen-derived danger signals and antigens. iNKT cells can serve both as helpers for effector B cells and negatively regulate autoreactive B cells. Specifically, iNKT cells drive B cell proliferation, class switch, and antibody production to induce primary antigen-specific immune responses. On the other hand, inflammasome-mediated activation drives accumulation of neutrophils, which license iNKT cells to negatively regulate autoreactive B cells via Fas ligand (FasL). This positions iNKT cells at an apex to support or inhibit B cell responses in inflammation. However, it is unknown which effector mechanism dominates in the face of cognate glycolipid activation during chronic inflammation, as might result from glycolipid vaccination or infection during chronic autoimmune disease. We stimulated iNKT cells by cognate glycolipid antigen α-galactosylceramide (αGalCer) and measured B cell activation during interleukin 18 (IL-18)-induced chronic inflammation. Moreover, glycolipid-activated iNKT cells increased the serum concentration of autoantibodies, frequency of germinal center (GC) B cells, and antigen-specific plasma cells induced during chronic IL-18-mediated inflammation, as compared with IL-18 alone. Further, activation of iNKT cells via cognate glycolipid during IL-18-mediated inflammation overrides the licensing function of neutrophils, instead inducing iNKT follicular helper (iNKTfh) cells that in turn promote autoimmunity. Thus, our data demonstrate that glycolipids which engage iNKT cells support antigen-specific B cell help during inflammasome-mediated inflammation.
Asunto(s)
Anticuerpos Antinucleares/inmunología , Autoinmunidad , Galactosilceramidas/inmunología , Inflamación/inmunología , Células T Asesinas Naturales/inmunología , Animales , Anticuerpos Antinucleares/sangre , Linfocitos B/inmunología , Enfermedad Crónica , Modelos Animales de Enfermedad , Femenino , Humanos , Inflamación/sangre , Inyecciones Intraperitoneales , Interleucina-18/administración & dosificación , Interleucina-18/inmunología , Masculino , Ratones , Ratones Transgénicos , Proteínas Recombinantes/administración & dosificación , Proteínas Recombinantes/inmunologíaRESUMEN
Tumor-associated macrophages (TAMs) can have protumor properties, including suppressing immune responses, promoting vascularization and, consequently, augmenting tumor progression. To stop TAM-mediated immunosuppression, we use a novel treatment by injecting antibodies specific for scavenger receptor MARCO, which is expressed on a specific subpopulation of TAMs in the tumor. We now report the location of this TAM as well as the pleiotropic mechanism of action of anti-MARCO antibody treatment on tumor progression and further show that this is potentially relevant to humans. Using specific targeting, we observed decreased tumor vascularization, a switch in the metabolic program of MARCO-expressing macrophages, and activation of natural killer (NK) cell killing through TNF-related apoptosis-inducing ligand (TRAIL). This latter activity reverses the effect of melanoma cell-conditioned macrophages in blocking NK activation and synergizes with T cell-directed immunotherapy, such as antibodies to PD-1 or PD-L1, to enhance tumor killing. Our study thus reveals an approach to targeting the immunosuppressive tumor microenvironment with monoclonal antibodies to enhance NK cell activation and NK cell-mediated killing. This can complement existing T cell-directed immunotherapy, providing a promising approach to combinatorial immunotherapy for cancer.
Asunto(s)
Protocolos de Quimioterapia Combinada Antineoplásica/farmacología , Células Asesinas Naturales/inmunología , Melanoma/tratamiento farmacológico , Receptores Inmunológicos/antagonistas & inhibidores , Macrófagos Asociados a Tumores/efectos de los fármacos , Animales , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Línea Celular Tumoral , Modelos Animales de Enfermedad , Femenino , Humanos , Inhibidores de Puntos de Control Inmunológico/farmacología , Inhibidores de Puntos de Control Inmunológico/uso terapéutico , Células Asesinas Naturales/metabolismo , Masculino , Melanoma/inmunología , Melanoma/patología , Ratones , Ratones Noqueados , Cultivo Primario de Células , Receptores Inmunológicos/genética , Receptores Inmunológicos/metabolismo , Ligando Inductor de Apoptosis Relacionado con TNF/metabolismo , Microambiente Tumoral/efectos de los fármacos , Microambiente Tumoral/inmunología , Macrófagos Asociados a Tumores/inmunología , Macrófagos Asociados a Tumores/metabolismoRESUMEN
BACKGROUND: Acute infection is a well-established risk factor of cardiovascular inflammation increasing the risk for a cardiovascular complication within the first weeks after infection. However, the nature of the processes underlying such aggravation remains unclear. Lipopolysaccharide derived from Gram-negative bacteria is a potent activator of circulating immune cells including neutrophils, which foster inflammation through discharge of neutrophil extracellular traps (NETs). Here, we use a model of endotoxinemia to link acute infection and subsequent neutrophil activation with acceleration of vascular inflammation Methods: Acute infection was mimicked by injection of a single dose of lipopolysaccharide into hypercholesterolemic mice. Atherosclerosis burden was studied by histomorphometric analysis of the aortic root. Arterial myeloid cell adhesion was quantified by intravital microscopy. RESULTS: Lipopolysaccharide treatment rapidly enhanced atherosclerotic lesion size by expansion of the lesional myeloid cell accumulation. Lipopolysaccharide treatment led to the deposition of NETs along the arterial lumen, and inhibition of NET release annulled lesion expansion during endotoxinemia, thus suggesting that NETs regulate myeloid cell recruitment. To study the mechanism of monocyte adhesion to NETs, we used in vitro adhesion assays and biophysical approaches. In these experiments, NET-resident histone H2a attracted monocytes in a receptor-independent, surface charge-dependent fashion. Therapeutic neutralization of histone H2a by antibodies or by in silico designed cyclic peptides enables us to reduce luminal monocyte adhesion and lesion expansion during endotoxinemia. CONCLUSIONS: Our study shows that NET-associated histone H2a mediates charge-dependent monocyte adhesion to NETs and accelerates atherosclerosis during endotoxinemia.
Asunto(s)
Aterosclerosis/metabolismo , Adhesión Celular/fisiología , Endotoxemia/metabolismo , Monocitos/metabolismo , Electricidad Estática , Animales , Aterosclerosis/inducido químicamente , Aterosclerosis/patología , Adhesión Celular/efectos de los fármacos , Endotoxemia/inducido químicamente , Endotoxemia/patología , Trampas Extracelulares/metabolismo , Humanos , Lipopolisacáridos/toxicidad , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Monocitos/efectos de los fármacos , Monocitos/patologíaRESUMEN
Vortex phase masks have been shown to be an efficient means to reduce the blinding stellar light in high-contrast imaging instruments. Once placed at the focal plane of the telescope, the helical phase ramp of a vortex phase mask diffracts the light of a bright on-axis source outside the re-imaged telescope pupil, while transmitting the light of a faint off-axis companion nearly unaffected. The Annular Groove Phase Mask (AGPM) is a broadband metasurface implementation of a vector vortex phase mask using the artificial birefringence of a circular subwavelength grating etched onto a diamond substrate. To date, the AGPM design has been optimized using rigorous coupled-wave analysis (RCWA), which is a valid tool to simulate periodic straight gratings. However, we have now reached a performance level where the curvature of the grating lines at the center becomes a limiting factor. Here, we use a finite-difference time-domain (FDTD) method to correctly describe the AGPM performance, including the effect of the curved grating close to its center. We confirm the validity of this simulation framework by comparing its predictions with experimental results obtained on our infrared coronagraphic test bench, and we show that RCWA fails at reproducing correctly the central AGPM performance, confirming the need for a full 3d simulation tool such as FDTD. Finally, we use FDTD to optimize the grating parameters at the AGPM center, and conclude with a new optimal design.
RESUMEN
The field of dielectric laser accelerators (DLA) garnered a considerable interest in the past six years as it offers novel opportunities in accelerator science and potentially transformative applications. Currently, the most widespread approach considers silicon-based structures due to their low absorption and high refractive index in the infrared spectral region and the well-developed silicon processing technology. In this paper we investigate a diamond as an alternative to silicon, mainly due to its considerably higher damage threshold. In particular, we find that our diamond grating allows a three times higher acceleration gradient (60â MeV/m) compared to silicon gratings designed for a similar electron energy. Using more complex geometries, GeV/m acceleration gradients are within reach for subrelativistic electrons.
RESUMEN
Marginal zone (MZ) B cells are innate-like B cells that produce polyreactive antibodies with an affinity for microbial molecular patterns and carbohydrate ligands. MZ B cells have been shown to be important in mediating immunity to various bacteria including Streptococcus pneumoniae and are also implicated in inflammatory syndromes including lupus erythematosus. The intestinal microbiota is responsible for producing short-chain fatty acids, which can regulate immune cell function by several mechanisms including ligation of the G-protein-coupled receptor (GPR)43. Herein, we show that MZ B cells express Gpr43 messenger RNA and that the absence of this receptor impacts on MZ B-cell surface marker expression and antibody production. In T-cell-independent responses to the hapten 4-hydroxy-3-nitrophenylacetic acid (NP), mice deficient in GPR43 displayed higher serum titers of NP-specific antibodies. Moreover, in response to a pneumococcal polysaccharide vaccine, GPR43-deficient mice developed robust serum antibody responses and had markedly increased numbers of splenic antibody-secreting cells, compared with control mice. Finally, serum immunoglobulin M autoantibodies to double-stranded DNA and phosphatidylcholine were increased in resting 10-15-week-old mice lacking GPR43. Taken together, mice lacking GPR43 have heightened antibody responses to T-cell-independent antigens, which may be a result of impaired regulation of MZ B cells.
Asunto(s)
Linfocitos B , Ácidos Grasos Volátiles , Animales , Células Productoras de Anticuerpos , Ratones , Ratones Endogámicos C57BL , Ratones NoqueadosRESUMEN
In systemic autoimmune diseases such as systemic lupus erythematosus (SLE), B cell tolerance is lost and there is a production of autoantibodies that drive pathology. The specificities of these antibodies are towards a wide range of autoantigens including proteins such as serum factors including cytokines as well as towards nucleic acids and modified glycolipids. It is known that endosomal pattern recognition receptors are involved in specific responses but if they drive specificity towards a specific group of autoantigens is not known. Here, we used syngeneic apoptotic cells alone to break B cell tolerance and investigated the antibody response in Unc93b1 mutant mice that lack signalling from the TLR3, TLR7 and TLR9 receptors. We found that specific B cell responses known from patients with SLE including antibodies towards Ro-52/60, La, cardiolipin as well as DNA were all significantly lower in the knockout mice. Thus, we found that endosomal TLR receptors were involved in break of tolerance and drive B cell responses for protein, nucleic acid and modified lipid antigens. This pinpoints these receptors as key drivers for the full range of antibody driven pathology in SLE and suggests that targeting of endosomal TLR driven responses will quench all B cell driven autoreactivity.
Asunto(s)
Apoptosis/inmunología , Autoantígenos/inmunología , Autoinmunidad , Linfocitos B/inmunología , Linfocitos B/metabolismo , Endosomas/metabolismo , Receptores de Reconocimiento de Patrones/metabolismo , Animales , Autoanticuerpos/inmunología , Biomarcadores , Modelos Animales de Enfermedad , Susceptibilidad a Enfermedades , Humanos , Lupus Eritematoso Sistémico/etiología , Lupus Eritematoso Sistémico/metabolismo , Lupus Eritematoso Sistémico/patología , RatonesRESUMEN
The role of diet in sarcopenia is unclear, and results from studies using dietary patterns (DP) are inconsistent. We assessed how adherences to a posteriori DP are associated with the prevalence of sarcopenia and its components 16 years later. Four DP were defined in the Uppsala Longitudinal Study of Adult Men at baseline (n 1133, average age 71 years). Among 257 men with information at follow-up, 19 % (n 50) had sarcopenia according to the European Working Group on sarcopenia in Older People 2 definition. Adherence to DP2 (mainly characterised by high intake of vegetables, green salad, fruit, poultry, rice and pasta) was non-linearly associated with sarcopenia; adjusted OR and 95 % CI for medium and high v. low adherence: 0·41 (0·17, 0·98) and 0·40 (0·17, 0·94). The OR per standard deviation (sd) higher adherence to DP2 was 0·70 (0·48, 1·03). Adjusted OR (95 % CI) for 1 sd higher adherence to DP1 (mainly characterised by high consumption of milk and cereals), DP3 (mainly characterised by high consumption of bread, cheese, marmalade, jam and sugar) and DP4 (mainly characterised by high consumption of potatoes, meat and egg and low consumption of fermented milk) were 1·04 (0·74, 1·46), 1·19 (0·71, 2·00) and 1·08 (0·77, 1·53), respectively. There were no clear associations between adherence to the DP and muscle strength, muscle mass, physical performance or sarcopenia using EWGSOP1 (sarcopenia n 54). Our results indicate that diet may be a potentially modifiable risk factor for sarcopenia in old Swedish men.
RESUMEN
Mid-infrared waveguide spectroscopy promises highly sensitive detection and characterization of organic molecules. Different material combinations for waveguides and cladding have been demonstrated with promising results, each with its own strengths and weaknesses in terms of sensitivity, transmission window and robustness. In this article we present a 5 µm thick diamond planar waveguide on aluminium nitride cladding, using a new fabrication and polishing method. Diamond has a very wide transmission window in the infrared, and its hardness and high chemical stability allows for chemistries and cleaning protocols that may damage other materials. With an aluminium nitride cladding the waveguide has a useable range between 1000 and 1900 cm-1, which we demonstrate using a tunable quantum cascade laser (QCL). This is a large improvement over silicon dioxide cladding. Compared to previously demonstrated free-standing diamond waveguides, the robustness of the sensor is greatly improved, which allows for a thinner diamond layer and increased sensitivity. The new waveguide was used in a QCL-based optical setup to detect acetone in deuterium oxide and isopropyl alcohol in water. The measurements showed higher sensitivity and lower noise level than previous demonstrations of mid-infrared diamond waveguides, resulting in a two orders of magnitude lower detectable concentration.
RESUMEN
Biodiversity offsetting is an increasingly applied tool aiming to compensate for environmental damage caused by exploitation projects. Critics, however, raise concerns over the purported effectiveness of offsetting and question the ethical underpinnings and implications of offsetting. These ethical dimensions have largely been overlooked in research, which may lead to offsetting systems that fail to respect the values intended to be safeguarded. To address these dimensions, 5 ethical objections in the scientific literature were identified: offsetting violates nature's intrinsic value; losses of nature cannot be compensated for by human interventions; too little is known to make adequate trades; offsetting impedes virtuous dispositions toward nature; and offsetting has negative justice implications. We examined these objections and arguments against them based on the ethical concepts of intrinsic and instrumental values, anthropocentrism, nonanthropocentrism, and deontological, consequentialist, and virtue-ethical paradigms. Both nonanthropocentric and anthropocentric concerns were expressed in deontological, consequential, and virtue-ethical framings. Objections mostly had a deontological or virtue-ethical basis, whereas counterarguments were based on consequential reasoning, but common ground in practice is often conceivable. Based on our findings, we formulated 10 recommendations for policy makers and 5 questions for practitioners to consider. We propose, for example, that policy makers clarify aims, legislate on no-go areas, and govern the use of multipliers. We suggest that practitioners consider, for instance, how to improve case-specific knowledge and promote learning and stakeholder engagement. We hope these recommendations and questions will encourage further discussion of the ethics of biodiversity offsets and ultimately strengthen the respect for biodiversity and human-welfare values at stake in offsetting projects.
Ética y Compensación de Biodiversidad Resumen La compensación de la biodiversidad es una herramienta que cada vez se aplica más a la búsqueda de la remuneración por el daño ambiental causado por proyectos de explotación. Sin embargo, hay quienes la critican basándose en la presunta efectividad de las compensaciones y cuestionan los sustentos éticos y las implicaciones de las compensaciones. Estas dimensiones éticas han sido ignoradas en la investigación, lo cual puede resultar en sistemas que no respetan los valores que se pretende salvaguardar. Para resolver estas dimensiones identificamos cinco objeciones éticas en la literatura: las compensaciones violan el valor intrínseco de la naturaleza; las pérdidas de la naturaleza no pueden compensarse mediante intervenciones humanas; se conoce muy poco para realizar intercambios adecuados; las compensaciones impiden las disposiciones virtuosas hacia la naturaleza; y las compensaciones tienen implicaciones de justicia negativa. Analizamos estas objeciones y los argumentos en su contra basados en los conceptos éticos de los valores intrínsecos e instrumentales, el antropocentrismo, el no antropocentrismo y los paradigmas deontológico, consecuencialista y de virtud ética. Tanto las preocupaciones antropocéntricas como las no antropocéntricas se expresaron mediante encuadres deontológicos, consecuencialistas y de virtud ética. Las objeciones principalmente tuvieron una base deontológica o de virtud ética, mientras que los contraargumentos estuvieron basados en el razonamiento consecuencial pero los puntos de coincidencia en la práctica con frecuencia son concebibles. Con base en nuestros hallazgos formulamos diez recomendaciones para los formuladores de políticas y cinco preguntas para los practicantes de la conservación para que las tomen a consideración. Por ejemplo, proponemos que los formuladores de políticas clarifiquen los objetivos, legislen las áreas prohibidas y determinen el uso de multiplicadores. Sugerimos que los practicantes consideren cómo mejorar el conocimiento específico por caso y promover el aprendizaje y la participación de los actores sociales. Esperamos que estas recomendaciones y preguntas fomenten una mayor discusión sobre la ética de las compensaciones de biodiversidad y, finalmente, fortalezcan el respeto por la biodiversidad y los valores de bienestar humano en juego en los proyectos de compensaciones.
Asunto(s)
Biodiversidad , Conservación de los Recursos Naturales , Humanos , Principios Morales , Justicia SocialRESUMEN
Extracellular vesicles (EV) are candidates for cancer immunotherapy because of their capacity to stimulate tumor-specific activity in vivo. However, clinical trials using peptide-loaded autologous EVs have so far only showed moderate T cell responses, suggesting a need for optimization of EV-induced immunity in humans. We previously demonstrated that induction of Ag-specific CD8+ T cells and antitumor responses to whole Ag were independent of MHC class I on EVs and hypothesized that multiple injections of allogeneic EVs could potentiate Ag-specific responses. In this study, we show that the allogeneic EV from mouse bone marrow-derived dendritic cells enhances Ag-specific CD8+ T cell, follicular helper T cell, and Ag-specific Ab responses. EV-injected mice demonstrated Ag-specific memory after 4 mo, with the highest Ab avidity in mice receiving double allogeneic EV injections. Reduced B16mOVA melanoma tumor growth was shown in all EV-injected groups. Our findings support the application of allogeneic EVs for therapeutic use in clinical studies in which an adaptive immune response is desired.
Asunto(s)
Vesículas Extracelulares/trasplante , Memoria Inmunológica/inmunología , Inmunoterapia/métodos , Melanoma Experimental/inmunología , Aloinjertos , Animales , Células de la Médula Ósea/inmunología , Células Dendríticas/inmunología , Vesículas Extracelulares/inmunología , Isoinjertos , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BLRESUMEN
Macrophages exist as innate immune subsets that exhibit phenotypic heterogeneity and functional plasticity. Their phenotypes are dictated by inputs from the tissue microenvironment. G-protein-coupled receptors are essential in transducing signals from the microenvironment, and heterotrimeric Gα signaling links these receptors to downstream effectors. Several Gαi-coupled G-protein-coupled receptors have been implicated in macrophage polarization. In this study, we use genetically modified mice to investigate the role of Gαi2 on inflammasome activity and macrophage polarization. We report that Gαi2 in murine bone marrow-derived macrophages (BMDMs) regulates IL-1ß release after activation of the NLRP3, AIM2, and NLRC4 inflammasomes. We show this regulation stems from the biased polarity of Gαi2 deficient (Gnai2 -/-) and RGS-insensitive Gαi2 (Gnai2 G184S/G184S) BMDMs. We determined that although Gnai2 G184S/G184S BMDMs (excess Gαi2 signaling) have a tendency toward classically activated proinflammatory (M1) phenotype, Gnai2-/- BMDMs (Gαi2 deficient) are biased toward alternatively activated anti-inflammatory (M2) phenotype. Finally, we find that Gαi2-deficient macrophages have increased Akt activation and IFN-ß production but defects in ERK1/2 and STAT3 activation after LPS stimulation. Gαi2-deficient macrophages also exhibit increased STAT6 activation after IL-4 stimulation. In summary, our data indicates that excess Gαi2 signaling promotes an M1 macrophage phenotype, whereas Gαi2 signaling deficiency promotes an M2 phenotype. Understanding Gαi2-mediated effects on macrophage polarization may bring to light insights regarding disease pathogenesis and the reprogramming of macrophages for the development of novel therapeutics.
Asunto(s)
Citocinas/biosíntesis , Subunidad alfa de la Proteína de Unión al GTP Gi2/inmunología , Inflamasomas/inmunología , Macrófagos/inmunología , Transducción de Señal/inmunología , Animales , Células Cultivadas , Subunidad alfa de la Proteína de Unión al GTP Gi2/deficiencia , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , FenotipoRESUMEN
Phosphorylcholine (PC) is an epitope on oxidized low-density lipoprotein (oxLDL), apoptotic cells and several pathogens like Streptococcus pneumoniae. Immunoglobulin M against PC (IgM anti-PC) has the ability to inhibit uptake of oxLDL by macrophages and increase clearance of apoptotic cells. From our genome-wide association studies (GWASs) in four European-ancestry cohorts, six single nucleotide polymorphisms (SNPs) in 11q24.1 were discovered (in 3002 individuals) and replicated (in 646 individuals) to be associated with serum level of IgM anti-PC (the leading SNP rs35923643-G, combined ß = 0.19, 95% confidence interval 0.13-0.24, P = 4.3 × 10-11). The haplotype tagged by rs35923643-G (or its proxy SNP rs735665-A) is also known as the top risk allele for chronic lymphocytic leukemia (CLL), and a main increasing allele for general IgM. By using summary GWAS results of IgM anti-PC and CLL in the polygenic risk score (PRS) analysis, PRS on the basis of IgM anti-PC risk alleles positively associated with CLL risk (explained 0.6% of CLL variance, P = 1.2 × 10-15). Functional prediction suggested that rs35923643-G might impede the binding of Runt-related transcription factor 3, a tumor suppressor playing a central role in the immune regulation of cancers. Contrary to the expectations from the shared genetics between IgM anti-PC and CLL, an inverse relationship at the phenotypic level was found in a nested case-control study (30 CLL cases with 90 age- and sex-matched controls), potentially reflecting reverse causation. The suggested function of the top variant as well as the phenotypic association between IgM anti-PC and CLL risk needs replication and motivates further studies.
Asunto(s)
Anticuerpos/sangre , Subunidad alfa 3 del Factor de Unión al Sitio Principal/genética , Inmunoglobulina M/genética , Leucemia Linfocítica Crónica de Células B/genética , Fosforilcolina/sangre , Adulto , Anciano , Anticuerpos/genética , Apoptosis/genética , Epítopos/sangre , Epítopos/genética , Epítopos/inmunología , Femenino , Estudio de Asociación del Genoma Completo , Haplotipos , Humanos , Inmunoglobulina M/sangre , Inmunoglobulina M/inmunología , Leucemia Linfocítica Crónica de Células B/sangre , Leucemia Linfocítica Crónica de Células B/inmunología , Lipoproteínas LDL/sangre , Lipoproteínas LDL/genética , Lipoproteínas LDL/inmunología , Macrófagos/inmunología , Masculino , Persona de Mediana Edad , Fosforilcolina/inmunología , Polimorfismo de Nucleótido Simple/genéticaRESUMEN
Natural killer T (NKT) cells recognize glycolipids presented on CD1d. They share features of adaptive T lymphocytes and innate NK cells, and mediate immunoregulatory functions via rapid production of cytokines. Invariant (iNKT) and diverse (dNKT) NKT cell subsets are defined by their TCR. The immunological role of dNKT cells, that do not express the invariant TCRα-chain used by iNKT cells, is less well explored than that of iNKT cells. Here, we investigated signals driving Toll-like receptor (TLR) ligand activation of TCR-transgenic murine dNKT cells. IFN-γ production by dNKT cells required dendritic cells (DC), cell-to-cell contact and presence of TLR ligands. TLR-stimulated DC activated dNKT cells to secrete IFN-γ in a CD1d-, CD80/86- and type I IFN-independent manner. In contrast, a requirement for IL-12p40, and a TLR ligand-selective dependence on IL-18 or IL-15 was observed. TLR ligand/DC stimulation provoked early secretion of pro-inflammatory cytokines by both CD62L+ and CD62L- dNKT cells. However, proliferation was limited. In contrast, TCR/co-receptor-mediated activation resulted in proliferation and delayed production of a broader cytokine spectrum preferentially in CD62L- dNKT cells. Thus, innate (TLR ligand/DC) and adaptive (TCR/co-receptor) stimulation of dNKT cells resulted in distinct cellular responses that may contribute differently to the formation of immune memory.
Asunto(s)
Inmunidad Adaptativa/inmunología , Citocinas/inmunología , Inmunidad Innata/inmunología , Linfocitos/inmunología , Células T Asesinas Naturales/inmunología , Animales , Antígenos CD1d/genética , Antígenos CD1d/inmunología , Antígenos CD1d/metabolismo , Proliferación Celular/genética , Células Cultivadas , Citocinas/metabolismo , Inmunidad Celular/inmunología , Ligandos , Linfocitos/metabolismo , Ratones Endogámicos C57BL , Ratones Noqueados , Ratones Transgénicos , Células T Asesinas Naturales/metabolismo , Receptores Toll-Like/inmunología , Receptores Toll-Like/metabolismoRESUMEN
Marginal zone (MZ) B cells reside in the splenic MZ and play important roles in T cell-independent humoral immune responses against blood-borne pathogens. IκBNS-deficient bumble mice exhibit a severe reduction in the MZ B compartment but regain an MZ B population with age and, thus, represent a valuable model to examine the biology of MZ B cells. In this article, we characterized the MZ B cell defect in further detail and investigated the nature of the B cells that appear in the MZ of aged bumble mice. Flow cytometry analysis of the splenic transitional B cell subsets demonstrated that MZ B cell development was blocked at the transitional-1 to transitional-2-MZ precursor stage in the absence of functional IκBNS. Immunohistochemical analysis of spleen sections from wild-type and bumble mice revealed no alteration in the cellular MZ microenvironment, and analysis of bone marrow chimeras indicated that the MZ B cell development defect in bumble mice was B cell intrinsic. Further, we demonstrate that the B cells that repopulate the MZ in aged bumble mice were distinct from age-matched wild-type MZ B cells. Specifically, the expression of surface markers characteristic for MZ B cells was altered and the L chain Igλ+ repertoire was reduced in bumble mice. Finally, plasma cell differentiation of sorted LPS-stimulated MZ B cells was impaired, and aged bumble mice were unable to respond to NP-Ficoll immunization. These results demonstrate that IκBNS is required for an intact MZ B cell compartment in C57BL/6 mice.
Asunto(s)
Subgrupos de Linfocitos B/inmunología , Subgrupos de Linfocitos B/metabolismo , Selección Clonal Mediada por Antígenos , Quinasa I-kappa B/deficiencia , Bazo/inmunología , Bazo/metabolismo , Factores de Edad , Animales , Antígenos T-Independientes/inmunología , Subgrupos de Linfocitos B/citología , Biomarcadores , Diferenciación Celular , Inmunoglobulina G/genética , Inmunoglobulina G/inmunología , Inmunofenotipificación , Lipopolisacáridos/inmunología , Ratones , Ratones Noqueados , FenotipoRESUMEN
Apoptotic cells drive innate regulatory responses that result in tolerogenic immunity. This is a critical aspect of cell physiology as apoptotic cells expose potentially dangerous nuclear antigens on the surface in apoptotic blebs, and failure in their recognition, phagocytosis, or destruction can cause dramatic autoimmunity in experimental models and is linked to development and progression of systemic pathology in human. The marginal zone is a specialized splenic environment that serves as a transitional site from circulation to peripheral lymphoid structures. The marginal zone serves a key role in trapping of particulates and initiation of innate responses against systemic microbial pathogens. However in recent years, it has become clear the marginal zone is also important for initiation of immune tolerance to apoptotic cells, driving a coordinated response involving multiple phagocyte and lymphocyte subsets. Recent reports linking defects in splenic macrophage function to systemic lupus erythematosus in a manner analogous to marginal zone macrophages in lupus-prone mice provide an impetus to better understand the mechanistic basis of the apoptotic cell response in the marginal zone and its general applicability to apoptotic cell-driven tolerance at other tissue sites. In this review, we discuss immune responses to apoptotic cells in the spleen in general and the marginal zone in particular, the relationship of these responses to autoimmune disease, and comparisons to apoptotic cell immunity in humans.