RESUMEN
Deposition of immune complexes (ICs) in tissues triggers acute inflammatory pathology characterized by massive neutrophil influx leading to edema and hemorrhage, and is especially associated with vasculitis of the skin, but the mechanisms that regulate this type III hypersensitivity process remain poorly understood. Here, using a combination of multiphoton intravital microscopy and genomic approaches, we re-examined the cutaneous reverse passive Arthus reaction and observed that IC-activated neutrophils underwent transmigration, triggered further IC formation, and transported these ICs into the interstitium, whereas neutrophil depletion drastically reduced IC formation and ameliorated vascular leakage in vivo. Thereafter, we show that these neutrophils expressed high levels of CXCL2, which further amplified neutrophil recruitment and activation in an autocrine and/or paracrine manner. Notably, CXCL1 expression was restricted to tissue-resident cell types, but IC-activated neutrophils may also indirectly, via soluble factors, modulate macrophage CXCL1 expression. Consistent with their distinct cellular origins and localization, only neutralization of CXCL2 but not CXCL1 in the interstitium effectively reduced neutrophil recruitment. In summary, our study establishes that neutrophils are able to self-regulate their own recruitment and responses during IC-mediated inflammation through a CXCL2-driven feed forward loop.
Asunto(s)
Complejo Antígeno-Anticuerpo/inmunología , Quimiocina CXCL2/metabolismo , Dermatitis/inmunología , Enfermedades del Complejo Inmune/inmunología , Neutrófilos/inmunología , Animales , Células Cultivadas , Quimiocina CXCL2/inmunología , Dermatitis/metabolismo , Modelos Animales de Enfermedad , Ensayo de Inmunoadsorción Enzimática , Femenino , Enfermedades del Complejo Inmune/fisiopatología , Mediadores de Inflamación/metabolismo , Macrófagos/inmunología , Masculino , Mastocitos/inmunología , Ratones , Ratones Endogámicos C57BL , Infiltración Neutrófila/inmunología , Neutrófilos/metabolismo , ARN Mensajero/análisisAsunto(s)
Células de Langerhans/citología , Piel/lesiones , Animales , Proteínas Bacterianas/metabolismo , Antígeno CD11c/metabolismo , Movimiento Celular , Dermis/citología , Dermis/metabolismo , Homeostasis , Inflamación , Rayos Láser , Proteínas Luminiscentes/metabolismo , Ganglios Linfáticos/patología , Ratones , Microscopía/métodos , Neutrófilos/citología , Neutrófilos/metabolismo , Piel/metabolismo , Piel/patologíaRESUMEN
Pivotal to brain development and function is an intact blood-brain barrier (BBB), which acts as a gatekeeper to control the passage and exchange of molecules and nutrients between the circulatory system and the brain parenchyma. The BBB also ensures homeostasis of the central nervous system (CNS). We report that germ-free mice, beginning with intrauterine life, displayed increased BBB permeability compared to pathogen-free mice with a normal gut flora. The increased BBB permeability was maintained in germ-free mice after birth and during adulthood and was associated with reduced expression of the tight junction proteins occludin and claudin-5, which are known to regulate barrier function in endothelial tissues. Exposure of germ-free adult mice to a pathogen-free gut microbiota decreased BBB permeability and up-regulated the expression of tight junction proteins. Our results suggest that gut microbiota-BBB communication is initiated during gestation and propagated throughout life.
Asunto(s)
Barrera Hematoencefálica , Intestinos/microbiología , Microbiota , Animales , Femenino , Ratones , Permeabilidad , Embarazo , Uniones Estrechas/metabolismoRESUMEN
Basophils, a rare leukocyte population in peripheral circulation, are conventionally identified as CD45(int) CD49b(+) FcεRI(+) cells. Here, we show that basophils from blood and several organs of naïve wild-type mice express CD41, the α subunit of α(IIb)ß3 integrin. CD41 expression on basophils is upregulated after in vivo IL-3 treatment and during infection with Nippostrongylus brasiliensis (Nb). Moreover, CD41 can be used as a reliable marker for basophils, circumventing technical difficulties associated with FcεRI for basophil identification in a Nb infection model. In vitro anti-IgE cross-linking and IL-3 basophil stimulation showed that CD41 upregulation positively correlates with augmented surface expression of CD200R and increased production of IL-4/IL-13, indicating that CD41 is a basophil activation marker. Furthermore, we found that infection with Plasmodium yoelii 17X (Py17x) induced a profound basophilia and using Mcpt8(DTR) reporter mice as a basophil-specific depletion model, we verified that CD41 can be used as a marker to track basophils in the steady state and during infection. During malarial infection, CD41 expression on basophils is negatively regulated by IFN-γ and positively correlates with increased basophil IL-4 production. In conclusion, we provide evidence that CD41 can be used as both an identification and activation marker for basophils during homeostasis and immune challenge.
Asunto(s)
Basófilos/inmunología , Malaria/inmunología , Nippostrongylus/inmunología , Plasmodium yoelii/inmunología , Glicoproteína IIb de Membrana Plaquetaria/inmunología , Infecciones por Strongylida/inmunología , Animales , Anticuerpos Antihelmínticos/inmunología , Basófilos/patología , Femenino , Inmunoglobulina E/inmunología , Interleucina-3/inmunología , Interleucina-4/inmunología , Malaria/patología , Glicoproteínas de Membrana/inmunología , Ratones , Ratones Endogámicos BALB C , Ratones Noqueados , Infecciones por Strongylida/patologíaRESUMEN
Malaria, a disease caused by the Plasmodium parasite, remains one of the most deadly infectious diseases known to mankind. The parasite has a complex life cycle, of which only the erythrocytic stage is responsible for the diverse pathologies induced during infection. To date, the disease mechanisms that underlie these pathologies are still poorly understood. In the case of infections caused by Plasmodium falciparum, the species responsible for most malaria related deaths, pathogenesis is thought to be due to the sequestration of infected red blood cells (IRBCs) in deep tissues. Other human and rodent malaria parasite species are also known to exhibit sequestration. Here, we review the different techniques that allow researchers to study how rodent malaria parasites modify their host cells, the distribution of IRBCs in vivo as well as the interactions between IRBCs and host tissues.
Asunto(s)
Mediciones Luminiscentes/métodos , Imagen por Resonancia Magnética/métodos , Microscopía/métodos , Plasmodium/citología , Tomografía de Emisión de Positrones/métodos , Animales , Plasmodium/fisiología , RoedoresRESUMEN
Blood neutrophil homeostasis is essential for successful host defense against invading pathogens. Circulating neutrophil counts are positively regulated by CXCR2 signaling and negatively regulated by the CXCR4-CXCL12 axis. In particular, G-CSF, a known CXCR2 signaler, and plerixafor, a CXCR4 antagonist, have both been shown to correct neutropenia in human patients. G-CSF directly induces neutrophil mobilization from the bone marrow (BM) into the blood, but the mechanisms underlying plerixafor-induced neutrophilia remain poorly defined. Using a combination of intravital multiphoton microscopy, genetically modified mice and novel in vivo homing assays, we demonstrate that G-CSF and plerixafor work through distinct mechanisms. In contrast to G-CSF, CXCR4 inhibition via plerixafor does not result in neutrophil mobilization from the BM. Instead, plerixafor augments the frequency of circulating neutrophils through their release from the marginated pool present in the lung, while simultaneously preventing neutrophil return to the BM. Our study demonstrates for the first time that drastic changes in blood neutrophils can originate from alternative reservoirs other than the BM, while implicating a role for CXCR4-CXCL12 interactions in regulating lung neutrophil margination. Collectively, our data provides valuable insights into the fundamental regulation of neutrophil homeostasis, which may lead to the development of improved treatment regimens for neutropenic patients.
Asunto(s)
Médula Ósea/metabolismo , Movimiento Celular/efectos de los fármacos , Compuestos Heterocíclicos/farmacología , Pulmón/citología , Neutrófilos/citología , Receptores CXCR4/antagonistas & inhibidores , Animales , Bencilaminas , Médula Ósea/efectos de los fármacos , Ciclamas , Factor Estimulante de Colonias de Granulocitos/farmacología , Proteínas Fluorescentes Verdes/metabolismo , Humanos , Recuento de Leucocitos , Macaca fascicularis , Ratones , Microscopía de Fluorescencia por Excitación Multifotónica , Muramidasa/metabolismo , Músculo Esquelético/citología , Mutación/genética , Circulación Pulmonar , Receptores CXCR4/genética , Receptores CXCR4/metabolismo , Receptores de Interleucina-8B/metabolismoRESUMEN
Dendritic cells (DCs) residing in the lung are known to acquire inhaled Ag and, after migration to the draining bronchial lymph node (brLN), to present it to naive T cells in an either tolerogenic or immunogenic context. To visualize endogenous lung-derived DCs, we applied fluorescent latex beads (LXs) intratracheally, thereby in vivo labeling the majority of phagocytic cells within the lung. Of note, LX-bearing cells subsequently arriving in the draining brLN were found to represent lung-derived migratory DCs. Imaging explanted brLN by two-photon laser-scanning microscopy, we quantitatively analyzed the migration and interaction behavior of naive CD4(+) T cells and endogenous, lung-derived DC presenting airway-delivered Ag under inflammatory or noninflammatory conditions. Ag-specific naive CD4(+) T cells engaged in stable as well as transient contacts with LX-bearing DCs in both situations and displayed similar overall motility kinetics, including a pronounced decrease in motility at 16-20 h after antigenic challenge. In contrast, the comparative analysis of T cell-DC cluster sizes as well as contact durations strongly suggests that lung-derived migratory DCs and naive CD4(+) T cells form more stable, long-lasting contacts under inflammatory conditions favoring the induction of respiratory immunity.
Asunto(s)
Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD4-Positivos/metabolismo , Comunicación Celular/inmunología , Células Dendríticas/inmunología , Células Dendríticas/metabolismo , Tolerancia Inmunológica , Inmunidad Innata , Pulmón/inmunología , Animales , Bronquios/inmunología , Bronquios/patología , Linfocitos T CD4-Positivos/patología , Comunicación Celular/genética , Inhibición de Migración Celular/genética , Inhibición de Migración Celular/inmunología , Células Dendríticas/patología , Modelos Animales de Enfermedad , Tolerancia Inmunológica/genética , Inmunidad Innata/genética , Inflamación/genética , Inflamación/inmunología , Inflamación/patología , Pulmón/citología , Pulmón/patología , Ganglios Linfáticos/inmunología , Ganglios Linfáticos/patología , Ratones , Ratones Endogámicos BALB C , Ratones Noqueados , Ratones Transgénicos , Ovalbúmina/administración & dosificación , Ovalbúmina/inmunologíaRESUMEN
Mucosal vaccination via the respiratory tract can elicit protective immunity in animal infection models, but the underlying mechanisms are still poorly understood. We show that a single intranasal application of the replication-deficient modified vaccinia virus Ankara, which is widely used as a recombinant vaccination vector, results in prominent induction of bronchus-associated lymphoid tissue (BALT). Although initial peribronchiolar infiltrations, characterized by the presence of dendritic cells (DCs) and few lymphocytes, can be found 4 d after virus application, organized lymphoid structures with segregated B and T cell zones are first observed at day 8. After intratracheal application, in vitro-differentiated, antigen-loaded DCs rapidly migrate into preformed BALT and efficiently activate antigen-specific T cells, as revealed by two-photon microscopy. Furthermore, the lung-specific depletion of DCs in mice that express the diphtheria toxin receptor under the control of the CD11c promoter interferes with BALT maintenance. Collectively, these data identify BALT as tertiary lymphoid structures supporting the efficient priming of T cell responses directed against unrelated airborne antigens while crucially requiring DCs for its sustained presence.