RESUMO
Neurokinin-1 receptor (NK1R) signaling can be immunomodulatory and it can lead to preferential transmigration of CD14+CD16+ monocytes across the blood brain barrier, potentially promoting the development of inflammatory neurological diseases, such as neuroHIV. To evaluate how NK1R signaling alters monocyte biology, RNA sequencing was used to define NK1R-mediated transcriptional changes in different monocyte subsets. The data show that NK1R activation induces a greater number of changes in CD14+CD16+ monocytes (152 differentially expressed genes), than in CD14+CD16- monocytes (36 genes), including increases in the expression of NF-κB and components of the NLRP3 inflammasome pathway. These results suggest that NK1R may alter the inflammatory state of CD14+CD16+ monocytes, influencing the development of neuroinflammation.
Assuntos
Inflamação/imunologia , Monócitos/imunologia , Receptores da Neurocinina-1/imunologia , Transdução de Sinais/imunologia , Adulto , Feminino , Proteínas Ligadas por GPI/imunologia , Humanos , Masculino , Pessoa de Meia-Idade , Monócitos/metabolismo , Receptores de IgG/imunologia , Receptores da Neurocinina-1/metabolismo , TranscriptomaRESUMO
Atherosclerosis is the main cause of many cardiovascular diseases. Endothelial dysfunction is recognized as an early event in the development of atherosclerosis. Many drugs have been studied to mitigate hyperlipidemia-induced endothelial injury. Studies have demonstrated that neuropeptide substance P (SP) and its preferred receptor neurokinin receptor 1 (NK-1R) are involved in the pathological progression of cardiovascular disease. In this study, we show that aprepitant, a selective NK-1R antagonist, possesses beneficial effects that protect endothelial cells from oxidized low-density lipoprotein (ox-LDL)-induced inflammatory response and injury. Our data demonstrate that NK-1R is expressed in both aortic and vein-originated endothelial cells and that ox-LDL treatment induces NK-1R expression. Treatment with aprepitant suppresses induction of endothelial vascular adhesion molecule (VCAM-1 and E-selectin) and cytokine by ox-LDL. The presence of aprepitant mitigates adhesion of monocytes to endothelial cells and the reduction in eNOS/NO triggered by ox-LDL. Mechanistically, we demonstrate that aprepitant suppresses ERK5-KLF2 axis activation. Silencing of KLF2 abolishes the inhibitory role of aprepitant on ox-LDL-induced inflammatory response, suggesting that its action is dependent on KLF2. Collectively, our data support that aprepitant exerts an anti-inflammatory effect. Further research is required to investigate the therapeutic potential of aprepitant in vascular inflammation resulting from atherosclerosis.
Assuntos
Células Endoteliais da Veia Umbilical Humana/imunologia , Fatores de Transcrição Kruppel-Like/imunologia , Lipoproteínas LDL/imunologia , Receptores da Neurocinina-1/imunologia , Adesão Celular/imunologia , Selectina E/imunologia , Células Endoteliais da Veia Umbilical Humana/patologia , Humanos , Inflamação/imunologia , Inflamação/patologia , Monócitos/imunologia , Monócitos/patologia , Células U937 , Molécula 1 de Adesão de Célula Vascular/imunologiaRESUMO
BACKGROUND: The tachykinin substance P (SP) is recognized to exacerbate inflammation at peripheral sites via its target receptor, neurokinin 1 receptor (NK-1R), expressed by leukocytes. More recently, SP/NK-1R interactions have been associated with severe neuroinflammation and neuronal damage. We have previously demonstrated that NK-1R antagonists can limit neuroinflammatory damage in a mouse model of bacterial meningitis. Furthermore, we have since shown that these agents can attenuate bacteria-induced neuronal and glial inflammatory mediator production in nonhuman primate (NHP) brain explants and isolated neuronal cells, and following in vivo infection. METHODS: In the present study, we have assessed the ability of NHP brain explants, primary human microglia and astrocytes, and immortalized human glial cell lines to express NK-1R isoforms. We have utilized RT-PCR, immunoblot analysis, immunofluorescent microscopy, and/or flow cytometric analysis, to quantify NK-1R expression in each, at rest, or following bacterial challenge. Furthermore, we have assessed the ability of human microglia to respond to SP by immunoblot analysis of NF-kB nuclear translocation and determined the ability of this neuropeptide to augment inflammatory cytokine release and neurotoxic mediator production by human astrocytes using an ELISA and a neuronal cell toxicity assay, respectively. RESULTS: We demonstrate that human microglial and astrocytic cells as well as NHP brain tissue constitutively express robust levels of the full-length NK-1R isoform. In addition, we demonstrate that the expression of NK-1R by human astrocytes can be further elevated following exposure to disparate bacterial pathogens or their components. Importantly, we have demonstrated that NK-1R is functional in both human microglia and astrocytes and show that SP can augment the inflammatory and/or neurotoxic immune responses of glial cells to disparate and clinically relevant bacterial pathogens. CONCLUSIONS: The robust constitutive and functional expression of the full-length NK-1R isoform by human microglia and astrocytes, and the ability of SP to augment inflammatory signaling pathways and mediator production by these cells, support the contention that SP/NK-1R interactions play a significant role in the damaging neuroinflammation associated with conditions such as bacterial meningitis.
Assuntos
Astrócitos/metabolismo , Encéfalo/metabolismo , Microglia/metabolismo , Receptores da Neurocinina-1/metabolismo , Substância P/metabolismo , Animais , Astrócitos/imunologia , Encéfalo/imunologia , Linhagem Celular , Humanos , Inflamação/imunologia , Inflamação/metabolismo , Macaca mulatta , Meningites Bacterianas/imunologia , Meningites Bacterianas/metabolismo , Microglia/imunologia , Técnicas de Cultura de Órgãos , Receptores da Neurocinina-1/imunologia , Substância P/imunologiaRESUMO
Neuroblastoma is the most common solid extracranial malignant tumor in children. Despite recent advances in the treatment of this heterogenous tumor with surgery and chemotherapy, the prognosis in advanced stages remains poor. Interestingly, neuroblastoma is one of the few solid tumors, to date, in which an effect for targeted immunotherapy has been proven in controlled clinical trials, giving hope for further advances in the treatment of this and other tumors by targeted therapy. A large array of novel therapeutic options for targeted therapy of neuroblastoma is on the horizon. To this repεrtoirε, the neurokinin-1 receptor (NK1R) system was recently added. The present article explores the most recent developments in targeting neuroblastoma cells via the NK1R and how this new knowledge could be helpful to create new anticancer therapies agains neuroblastoma and other cancers.
Assuntos
Antineoplásicos/farmacologia , Imunoterapia , Terapia de Alvo Molecular , Neuroblastoma/terapia , Receptores da Neurocinina-1/química , Criança , Humanos , Neuroblastoma/imunologia , Receptores da Neurocinina-1/imunologiaRESUMO
Substance P neuropeptide and its receptor, neurokinin-1 receptor (NK1R), are reported to present on the ocular surface. In this study, mice lacking functional NK1R exhibited an excessive desquamation of apical corneal epithelial cells in association with an increased epithelial cell proliferation and increased epithelial cell density, but decreased epithelial cell size. The lack of NK1R also resulted in decreased density of corneal nerves, corneal epithelial dendritic cells (DCs), and a reduced volume of basal tears. Interestingly, massive accumulation of CD11c+CD11b+ conventional DCs was noted in the bulbar conjunctiva and near the limbal area of corneas from NK1R-/- mice. After ocular HSV-1 infection, the number of conventional DCs and neutrophils infiltrating the infected corneas was significantly higher in NK1R-/- than C57BL/6J mice. This was associated with an increased viral load in infected corneas of NK1R-/- mice. As a result, the number of IFN-γ-secreting virus-specific CD4 T cells in the draining lymph nodes of NK1R-/- mice was much higher than in infected C57BL/6J mice. An increased number of CD4 T cells and mature neutrophils (CD11b+Ly6ghigh) in the inflamed corneas of NK1R-/- mice was associated with an early development of severe herpes stromal keratitis. Collectively, our results show that the altered corneal biology of uninfected NK1R-/- mice along with an enhanced immunological response after ocular HSV-1 infection causes an early development of herpes stromal keratitis in NK1R-/- mice.
Assuntos
Córnea/imunologia , Córnea/patologia , Herpesvirus Humano 1/imunologia , Ceratite Herpética/virologia , Receptores da Neurocinina-1/fisiologia , Animais , Linfócitos T CD4-Positivos/imunologia , Túnica Conjuntiva/imunologia , Túnica Conjuntiva/patologia , Túnica Conjuntiva/virologia , Córnea/virologia , Células Dendríticas/imunologia , Herpesvirus Humano 1/fisiologia , Homeostase , Interferon gama/imunologia , Ceratite Herpética/imunologia , Ceratite Herpética/fisiopatologia , Linfonodos/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Neutrófilos/imunologia , Receptores da Neurocinina-1/deficiência , Receptores da Neurocinina-1/imunologia , Carga ViralRESUMO
BACKGROUND: The need for new options for chronic lung diseases promotes the research on stem cells for lung repair. Bone marrow-derived mesenchymal stem cells (MSCs) can modulate lung inflammation, but the data on cellular processes involved in early airway remodeling and the potential involvement of neuropeptides are scarce. OBJECTIVES: To elucidate the mechanisms by which local administration of MSCs interferes with pathophysiological features of airway hyperresponsiveness in an animal model. METHODS: GFP-tagged mouse MSCs were intratracheally delivered in the ovalbumin mouse model with subsequent functional tests, the analysis of cytokine levels, neuropeptide expression and histological evaluation of MSCs fate and airway pathology. Additionally, MSCs were exposed to pro-inflammatory factors in vitro. RESULTS: Functional improvement was observed after MSC administration. Although MSCs did not adopt lung cell phenotypes, cell therapy positively affected airway remodeling reducing the hyperplastic phase of the gain in bronchial smooth muscle mass, decreasing the proliferation of epithelium in which mucus metaplasia was also lowered. Decrease of interleukin-4, interleukin-5, interleukin-13 and increase of interleukin-10 in bronchoalveolar lavage was also observed. Exposed to pro-inflammatory cytokines, MSCs upregulated indoleamine 2,3-dioxygenase. Moreover, asthma-related in vivo upregulation of pro-inflammatory neurokinin 1 and neurokinin 2 receptors was counteracted by MSCs that also determined a partial restoration of VIP, a neuropeptide with anti-inflammatory properties. CONCLUSION: Intratracheally administered MSCs positively modulate airway remodeling, reduce inflammation and improve function, demonstrating their ability to promote tissue homeostasis in the course of experimental allergic asthma. Because of a limited tissue retention, the functional impact of MSCs may be attributed to their immunomodulatory response combined with the interference of neuropeptide system activation and tissue remodeling.
Assuntos
Transplante de Células-Tronco Mesenquimais , Células-Tronco Mesenquimais/imunologia , Receptores da Neurocinina-1/imunologia , Receptores da Neurocinina-2/imunologia , Hipersensibilidade Respiratória/terapia , Animais , Líquido da Lavagem Broncoalveolar/química , Líquido da Lavagem Broncoalveolar/imunologia , Expressão Gênica , Genes Reporter , Proteínas de Fluorescência Verde/genética , Proteínas de Fluorescência Verde/metabolismo , Indolamina-Pirrol 2,3,-Dioxigenase/genética , Indolamina-Pirrol 2,3,-Dioxigenase/imunologia , Interleucina-10/genética , Interleucina-10/imunologia , Interleucina-13/genética , Interleucina-13/imunologia , Interleucina-4/genética , Interleucina-4/imunologia , Interleucina-5/genética , Interleucina-5/imunologia , Intubação Intratraqueal , Pulmão/imunologia , Pulmão/patologia , Células-Tronco Mesenquimais/citologia , Camundongos , Camundongos Endogâmicos BALB C , Ovalbumina , Receptores da Neurocinina-1/genética , Receptores da Neurocinina-2/genética , Hipersensibilidade Respiratória/induzido quimicamente , Hipersensibilidade Respiratória/imunologia , Hipersensibilidade Respiratória/patologiaAssuntos
Apresentação de Antígeno , Células Dendríticas/imunologia , Neurocinina A/imunologia , Receptores da Neurocinina-1/imunologia , Receptores da Neurocinina-2/imunologia , Substância P/imunologia , Alveolite Alérgica Extrínseca/imunologia , Antígenos de Superfície/imunologia , Asma/imunologia , Diferenciação Celular/efeitos dos fármacos , Citocinas/imunologia , Células Dendríticas/citologia , Células Dendríticas/efeitos dos fármacos , Humanos , Antagonistas dos Receptores de Neurocinina-1/farmacologia , Poli I-C/farmacologia , Receptores da Neurocinina-2/antagonistas & inibidoresRESUMO
Cortical interneurons, immunoreactive for neuronal nitric oxide synthase (nNOS) and the receptor NK1, express the functional activity marker Fos selectively during sleep. NREM sleep 'pressure' is hypothesized to accumulate during waking and to dissipate during sleep. We reported previously that the proportion of Fos(+) cortical nNOS/NK1 neurons is correlated with established electrophysiological markers of sleep pressure. As these markers covary with the amount of NREM sleep, it remained unclear whether cortical nNOS/NK1 neurons are activated to the same degree throughout NREM sleep or whether the extent of their activation is related to the sleep pressure that accrued during the prior waking period. To distinguish between these possibilities, we used hypnotic medications to control the amount of NREM sleep in rats while we varied prior wake duration and the resultant sleep pressure. Drug administration was preceded by 6 h of sleep deprivation (SD) ('high sleep pressure') or undisturbed conditions ('low sleep pressure'). We find that the proportion of Fos(+) cortical nNOS/NK1 neurons was minimal when sleep pressure was low, irrespective of the amount of time spent in NREM sleep. In contrast, a large proportion of cortical nNOS/NK1 neurons was Fos(+) when an equivalent amount of sleep was preceded by SD. We conclude that, although sleep is necessary for cortical nNOS/NK1 neuron activation, the proportion of cells activated is dependent upon prior wake duration.
Assuntos
Córtex Cerebral/fisiologia , Homeostase/fisiologia , Neurônios/fisiologia , Óxido Nítrico Sintase Tipo I/fisiologia , Receptores da Neurocinina-1/fisiologia , Fases do Sono/fisiologia , Acetamidas/farmacologia , Animais , Córtex Cerebral/efeitos dos fármacos , Isoquinolinas/farmacologia , Masculino , Neurônios/imunologia , Óxido Nítrico Sintase Tipo I/imunologia , Piridinas/farmacologia , Ratos , Receptores da Neurocinina-1/imunologia , Privação do Sono/fisiopatologia , Fases do Sono/efeitos dos fármacos , ZolpidemRESUMO
TRPM8 is the molecular sensor for cold; however, the physiological role of TRPM8+ neurons at mucosal surfaces is unclear. Here we evaluated the distribution and peptidergic properties of TRPM8+ fibers in naive and inflamed colons, as well as their role in mucosal inflammation. We found that Trpm8(-/-) mice were hypersusceptible to dextran sodium sulfate (DSS)-induced colitis, and that Trpm8(-/-) CD11c+ DCs (dendritic cells) showed hyperinflammatory responses to toll-like receptor (TLR) stimulation. This was phenocopied in calcitonin gene-related peptide (CGRP) receptor-deficient mice, but not in substance P receptor-deficient mice, suggesting a functional link between TRPM8 and CGRP. The DSS phenotype of CGRP receptor-deficient mice could be adoptively transferred to wild-type (WT) mice, suggesting that CGRP suppresses the colitogenic activity of bone marrow-derived cells. TRPM8+ mucosal fibers expressed CGRP in human and mouse colon. Furthermore, neuronal CGRP contents were increased in colons from naive and DSS-treated Trpm8(-/-) mice, suggesting deficient CGRP release in the absence of TRPM8 triggering. Finally, treatment of Trpm8(-/-) mice with CGRP reversed their hyperinflammatory phenotype. These results suggest that TRPM8 signaling in mucosal sensory neurons is indispensable for the regulation of innate inflammatory responses via the neuropeptide CGRP.
Assuntos
Peptídeo Relacionado com Gene de Calcitonina/imunologia , Colite/imunologia , Imunidade Inata , Mucosa Intestinal/imunologia , Células Receptoras Sensoriais/imunologia , Canais de Cátion TRPM/imunologia , Animais , Peptídeo Relacionado com Gene de Calcitonina/deficiência , Peptídeo Relacionado com Gene de Calcitonina/genética , Colite/induzido quimicamente , Colite/genética , Colite/patologia , Colo/imunologia , Colo/patologia , Células Dendríticas/imunologia , Células Dendríticas/patologia , Sulfato de Dextrana , Células Epiteliais/imunologia , Células Epiteliais/patologia , Feminino , Regulação da Expressão Gênica , Humanos , Imunidade nas Mucosas , Mucosa Intestinal/patologia , Masculino , Camundongos Endogâmicos C57BL , Camundongos Knockout , Fenótipo , Receptores da Neurocinina-1/deficiência , Receptores da Neurocinina-1/genética , Receptores da Neurocinina-1/imunologia , Células Receptoras Sensoriais/patologia , Transdução de Sinais , Canais de Cátion TRPM/deficiência , Canais de Cátion TRPM/genéticaRESUMO
The substance P neurokinin 1 receptor (NK1R) regulates motility, secretion, inflammation and pain in the intestine. The distribution of the NK1R is a key determinant of the functional effects of substance P in the gut. Information regarding the distribution of NK1R in subtypes of mouse enteric neurons is lacking and is the focus of the present study. NK1R immunoreactivity (NK1R-IR) is examined in whole-mount preparations of the mouse distal colon by indirect immunofluorescence and confocal microscopy. The distribution of NK1R-IR within key functional neuronal subclasses was determined by using established neurochemical markers. NK1R-IR was expressed by a subpopulation of myenteric and submucosal neurons; it was mainly detected in large multipolar myenteric neurons and was colocalized with calcitonin gene-related peptide, neurofilament M, choline acetyltransferase and calretinin. The remaining NK1R-immunoreactive neurons were positive for nitric oxide synthase. NK1R was expressed by most of the submucosal neurons and was exclusively co-expressed with vasoactive intestinal peptide, with no overlap with choline acetyltransferase. Treatment with substance P resulted in the concentration-dependent internalisation of NK1R from the cell surface into endosome-like structures. Myenteric NK1R was mainly expressed by intrinsic primary afferent neurons, with minor expression by descending interneurons and inhibitory motor neurons. Submucosal NK1R was restricted to non-cholinergic secretomotor neurons. These findings highlight key differences in the neuronal distribution of NK1R-IR between the mouse, rat and guinea-pig, with important implications for the functional role of NK1R in regulating intestinal motility and secretion.
Assuntos
Colo/inervação , Sistema Nervoso Entérico/metabolismo , Receptores da Neurocinina-1/metabolismo , Substância P/metabolismo , Animais , Anticorpos/imunologia , Calbindina 2/metabolismo , Peptídeo Relacionado com Gene de Calcitonina/metabolismo , Colina O-Acetiltransferase/biossíntese , Colo/metabolismo , Feminino , Técnica Indireta de Fluorescência para Anticorpo , Trato Gastrointestinal/inervação , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Proteínas de Neurofilamentos/metabolismo , Óxido Nítrico Sintase/metabolismo , Receptores da Neurocinina-1/biossíntese , Receptores da Neurocinina-1/imunologia , Peptídeo Intestinal Vasoativo/biossínteseRESUMO
In this issue of Blood, Janelsins et al report that substance P (SP)treated dendritic cells (DCs) efficiently home to lymph nodes, where they induce inflammatory DCs to produce interleukin-12 (IL-12), thereby promoting type 1 polarized immunity.
Assuntos
Células Dendríticas/imunologia , Imunidade Celular/imunologia , Interleucina-12/imunologia , Receptores da Neurocinina-1/imunologia , AnimaisRESUMO
Substance-P and hemokinin-1 are proinflammatory neuropeptides with potential to promote type 1 immunity through agonistic binding to neurokinin-1 receptor (NK1R). Dendritic cells (DCs) are professional antigen-presenting cells that initiate and regulate the outcome of innate and adaptive immune responses. Immunostimulatory DCs are highly desired for the development of positive immunization techniques. DCs express functional NK1R; however, regardless of their potential DC-stimulatory function, the ability of NK1R agonists to promote immunostimulatory DCs remains unexplored. Here, we demonstrate that NK1R signaling activates therapeutic DCs capable of biasing type 1 immunity by inhibition of interleukin-10 (IL-10) synthesis and secretion, without affecting their low levels of IL-12 production. The potent type 1 effector immune response observed following cutaneous administration of NK1R-signaled DCs required their homing in skin-draining lymph nodes (sDLNs) where they induced inflammation and licensed endogenous-conventional sDLN-resident and -recruited inflammatory DCs to secrete IL-12. Our data demonstrate that NK1R signaling promotes immunostimulatory DCs, and provide relevant insight into the mechanisms used by neuromediators to regulate innate and adaptive immune responses.
Assuntos
Células Dendríticas/imunologia , Imunidade Celular/imunologia , Interleucina-12/imunologia , Receptores da Neurocinina-1/imunologia , Animais , Células da Medula Óssea/imunologia , Células da Medula Óssea/metabolismo , Células Cultivadas , Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/imunologia , Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/metabolismo , Células Dendríticas/metabolismo , Células Dendríticas/transplante , Citometria de Fluxo , Imunização/métodos , Imunofenotipagem , Interleucina-10/imunologia , Interleucina-10/metabolismo , Interleucina-12/genética , Interleucina-12/metabolismo , Alvo Mecanístico do Complexo 2 de Rapamicina , Camundongos , Camundongos da Linhagem 129 , Camundongos Endogâmicos C57BL , Camundongos Knockout , Camundongos Transgênicos , Complexos Multiproteicos/imunologia , Complexos Multiproteicos/metabolismo , Receptores da Neurocinina-1/agonistas , Receptores da Neurocinina-1/metabolismo , Transdução de Sinais/imunologia , Serina-Treonina Quinases TOR/imunologia , Serina-Treonina Quinases TOR/metabolismoRESUMO
To fulfill its immunologic functions, the skin is richly populated with dendritic cells (DCs), the most potent professional Ag-processing and Ag-presenting cells of the immune system. The immune-stimulatory and tolerogenic functions of skin DCs are regulated by the immune and neuroendocrine systems. Pro-inflammatory neuropeptides like substance P and calcitonin gene-related peptide secreted by cutaneous Aδ and C nerve fibers are the main initiators of neuro-inflammatory responses in the skin. Conversely, anti-inflammatory neuropeptides like the products of cleavage of proopiomelanocortin (α-melanocyte-stimulating hormone and adrenocorticotropic hormone) negatively regulate immune responses in the skin. Likewise, the control of immune responses against stimuli applied to the skin depends on the balance between the release and neutralization of pro-inflammatory and anti-inflammatory neuropeptides, a matter that has been somehow overlooked by the immunology field until recently.
Assuntos
Receptores da Neurocinina-1/imunologia , Receptores da Neurocinina-1/metabolismo , Dermatopatias/imunologia , Animais , Doença Crônica , Células Dendríticas/imunologia , Células Dendríticas/metabolismo , Humanos , Sistema Imunitário/imunologia , Sistema Imunitário/metabolismo , Inflamação/imunologia , Mastócitos/imunologia , Mastócitos/metabolismo , Sistema Nervoso/imunologia , Sistema Nervoso/metabolismo , Neuropeptídeos/imunologia , Neuropeptídeos/metabolismo , Receptores da Neurocinina-1/agonistas , Receptores de Taquicininas/metabolismo , Transdução de Sinais/imunologia , Pele/imunologia , Linfócitos T/imunologia , Linfócitos T/metabolismo , Taquicininas/imunologia , Taquicininas/metabolismoRESUMO
Present in vitro investigation for the first time in ectotherms demonstrated the immunomodulatory role of substance P in the wall lizard Hemidactylus flaviviridis. Substance P inhibited the percentage phagocytosis and phagocytic index of lizard splenic phagocytes. Inhibitory effect of substance P was completely blocked by NK-1 receptor antagonist spantide I, indicating the NK-1 receptor mediated action. Further, NK-1 receptor-coupled downstream signaling cascade involved in controlling phagocytosis was explored using inhibitors of adenylate cyclase (SQ 22536) and protein kinase A (H-89). Both the inhibitors, in a concentration-related manner decreased the suppressive effect of substance P on phagocytosis. In addition, substance P treatment caused an increase in intracellular cAMP level in splenic phagocytes. Taken together, it can be suggested that substance P via NK-1 receptor-coupled AC-cAMP-PKA pathway modulated the phagocytic activity of splenic phagocytes in wall lizards.
Assuntos
Lagartos/imunologia , Substância P/imunologia , Adenina/análogos & derivados , Adenina/farmacologia , Animais , Células Cultivadas , AMP Cíclico/análise , AMP Cíclico/metabolismo , Inibidores Enzimáticos/farmacologia , Feminino , Fatores Imunológicos/metabolismo , Fatores Imunológicos/farmacologia , Isoquinolinas/farmacologia , Antagonistas dos Receptores de Neurocinina-1 , Fagócitos/efeitos dos fármacos , Fagócitos/metabolismo , Fagocitose/efeitos dos fármacos , Fagocitose/imunologia , Receptores da Neurocinina-1/imunologia , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/imunologia , Baço/efeitos dos fármacos , Baço/imunologia , Substância P/análogos & derivados , Substância P/farmacologia , Sulfonamidas/farmacologiaRESUMO
Caveolin (Cav)-1 is an integral membrane protein of caveolae playing a crucial role in various signal transduction pathways. Caveolae represent the sites for calcium entry and storage especially in smooth muscle cells (SMC) and interstitial cells of Cajal (ICC). Cav-1(-/-) mice lack caveolae and show abnormalities in pacing and contractile activity of the small intestine. Presently, we investigated, by transmission electron microscopy (TEM) and immunohistochemistry, whether the absence of Cav-1 in Cav-1(-/-) mouse small intestine affects ICC, SMC and neuronal morphology, the expression of NK1 and NK2 receptors, and of Ano1 (also called Dog1 or TMEM16A), an essential molecule for slow wave activity in gastrointestinal muscles. ICC were also labelled with c-Kit and tachykinergic neurons with Substance P (SP). In Cav-1(-/-) mice: (i) ICC were Ano1-negative but maintained c-Kit expression, (ii) NK1 and NK2 receptor immunoreactivity was more intense and, in the SMC, mainly intracytoplasmatic, (iii) SP-immunoreactivity was significantly reduced. Under TEM: (i) ICC, SMC and telocytes lacked typical caveolae but had few and large flask-shaped vesicles we called large-sized caveolae; (ii) SMC and ICC contained an extraordinary high number of mitochondria, (iii) neurons were unchanged. To maintain intestinal motility, loss of caveolae and reduced calcium availability in Cav-1-knockout mice seem to be balanced by a highly increased number of mitochondria in ICC and SMC. Loss of Ano-1 expression, decrease of SP content and consequently overexpression of NK receptors suggest that all these molecules are Cav-1-associated proteins.
Assuntos
Caveolina 1/deficiência , Canais de Cloreto/metabolismo , Íleo/metabolismo , Íleo/ultraestrutura , Miócitos de Músculo Liso/metabolismo , Miócitos de Músculo Liso/ultraestrutura , Receptores da Neurocinina-1/biossíntese , Receptores da Neurocinina-2/biossíntese , Substância P/metabolismo , Animais , Anoctamina-1 , Cavéolas/metabolismo , Cavéolas/ultraestrutura , Caveolina 1/genética , Caveolina 1/metabolismo , Motilidade Gastrointestinal , Íleo/citologia , Íleo/imunologia , Imuno-Histoquímica , Células Intersticiais de Cajal/metabolismo , Camundongos , Camundongos Knockout , Microscopia Eletrônica de Transmissão , Mitocôndrias Musculares/ultraestrutura , Contração Muscular , Neurônios/citologia , Neurônios/metabolismo , Neurônios/ultraestrutura , Proteínas Proto-Oncogênicas c-kit/biossíntese , Proteínas Proto-Oncogênicas c-kit/metabolismo , Receptores da Neurocinina-1/imunologia , Receptores da Neurocinina-2/imunologia , Transdução de Sinais , Substância P/imunologiaRESUMO
The neuropeptide substance P (SP) can regulate a number of immunological functions in vitro and in vivo and may regulate natural killer (NK) cell activity. Here, we investigated whether SP has a role in regulating NK92-MI cell function in vitro, and how it influences NK cell activity. We found that SP dose dependently increased the cytotoxicity of NK92-MI cells and had a maximal effect at a concentration of 10(-12) and 10(-10) m. Furthermore, the expression of cytotoxic-associated molecules (perforin, granzyme) and activating receptor NKp46 [a member of natural cytotoxicity receptors (NCRs)] was observed to be upregulated by SP at optimal concentration, at which SP enhanced the cytotoxicity of NK92-MI cells. Neurokinin-1 receptor (NK-1R), a functional receptor of SP, was found on NK92-MI cells, and the observed effects of SP on NK92-MI cells could be more partially blocked by an NK-1R antagonist. Our data suggest that SP induces NK92-MI cell cytotoxicity by directly increasing the expression of cytotoxic granules and upregulates NK92-MI cell receptor-mediated functions indirectly. Thus, SP may regulate NK cell function mainly through NK-1R.
Assuntos
Citotoxicidade Imunológica , Células Matadoras Naturais/imunologia , Substância P/imunologia , Linhagem Celular , Granzimas/biossíntese , Granzimas/imunologia , Humanos , Células Matadoras Naturais/efeitos dos fármacos , Receptor 1 Desencadeador da Citotoxicidade Natural/biossíntese , Receptor 1 Desencadeador da Citotoxicidade Natural/imunologia , Antagonistas dos Receptores de Neurocinina-1 , Perforina/biossíntese , Perforina/imunologia , Receptores da Neurocinina-1/imunologia , Substância P/farmacologia , Regulação para Cima/efeitos dos fármacos , Regulação para Cima/imunologiaRESUMO
Transient receptor potential vanilloid 1 (TRPV1), neurokinin 1 (NK1) receptor and substance P (SP) immunoreactivity (-ir) and mRNA in the rat lumbosacral spinal cord and urinary bladder were measured 24h after s.c. injection of the vanilloids, capsaicin (50mg/kg) and resiniferatoxin (RTX, 100µg/kg), or vehicle (10% ethanol/10% Tween 80/saline). In the spinal cord, capsaicin significantly reduced TRPV1 and SP-ir (40-45%) in laminae I/II compared to controls, while RTX produced decreases of ~35%. NK1-ir in the spinal cord was unaffected by both vanilloid treatments. In the bladder, SP-ir was reduced in urothelial cells of some capsaicin- and RTX-treated rats, while SP-ir in the suburothelium and muscularis was significantly reduced by RTX. A significant increase in NK1-ir was observed in the urothelium and muscularis after capsaicin administration. Capsaicin significantly increased SP mRNA in the spinal cord, and TRPV1 and SP mRNA in the bladder, whereas RTX increased TRPV1, SP and NK1 mRNA in the spinal cord, and TRPV1 and SP mRNA in the bladder. These data suggest that stimulation of TRPV1 by low dose vanilloid administration can rapidly (within 24h) alter both transcription and translation of TRPV1 channels, SP and NK1 receptors in the rat urinary bladder and spinal cord.
Assuntos
Capsaicina/administração & dosagem , Diterpenos/administração & dosagem , Receptores da Neurocinina-1/metabolismo , Medula Espinal/metabolismo , Substância P/metabolismo , Canais de Cátion TRPV/metabolismo , Bexiga Urinária/metabolismo , Animais , Capsaicina/farmacologia , Diterpenos/farmacologia , Relação Dose-Resposta a Droga , Vértebras Lombares/metabolismo , Masculino , RNA Mensageiro/metabolismo , Ratos , Ratos Wistar , Receptores da Neurocinina-1/genética , Receptores da Neurocinina-1/imunologia , Sacro/metabolismo , Substância P/genética , Substância P/imunologia , Canais de Cátion TRPV/genética , Canais de Cátion TRPV/imunologiaRESUMO
Subepithelial fibroblasts of the intestinal villi, which form a contractile cellular network beneath the epithelium, are in close contact with epithelial cells, nerve varicosities, capillaries, smooth muscles and immune cells, and secrete extracellular matrix molecules, growth factors and cytokines, etc. Cultured subepithelial fibroblasts of the rat duodenal villi display various receptors such as endothelins, ATP, substance-P and bradykinin, and release ATP in response to mechanical stimulation. In this study, the presence of functional NK1 receptors (NK1R) was pharmacologically confirmed in primary culture by Ca(2+) measurement, and the effects of substance-P were measured in an acute preparation of epithelium-free duodenal villi from 2- to 3-week-old rats using a two-photon laser microscope. Substance-P elicited an increase in the intracellular Ca(2+) concentration and contraction of the subepithelial fibroblasts in culture and the isolated villi. The localization of NK1R and substance-P in the villi was examined by light and electron microscopic immunohistochemistry. NK1R-like immunoreactivity was intensely localized on the plasma membrane of villous subepithelial fibroblasts in 10-day- to 4-week-old rats and mice and was decreased or absent in adulthood. The pericryptal fibroblasts of the small and large intestine were NK1R immuno-negative. These villous subepithelial fibroblasts form synapse-like structures with both substance-P-immunopositive and -immunonegative nerve varicosities. Here, we propose that the mutual interaction between villous subepithelial fibroblasts and afferent neurons via substance-P and ATP plays important roles in the maturation of the structure and function of the small intestine.
Assuntos
Fibroblastos/citologia , Mucosa Intestinal/citologia , Receptores da Neurocinina-1/metabolismo , Substância P/metabolismo , Trifosfato de Adenosina/metabolismo , Animais , Cálcio/metabolismo , Sinalização do Cálcio , Membrana Celular/efeitos dos fármacos , Membrana Celular/metabolismo , Células Cultivadas , Feminino , Fibroblastos/efeitos dos fármacos , Fibroblastos/metabolismo , Mucosa Intestinal/efeitos dos fármacos , Mucosa Intestinal/metabolismo , Intestino Delgado/citologia , Intestino Delgado/efeitos dos fármacos , Intestino Delgado/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Neurônios Aferentes/metabolismo , Ratos , Ratos Mutantes , Ratos Wistar , Receptores da Neurocinina-1/imunologia , Substância P/farmacologiaRESUMO
Recent studies suggest a link between neuropsychiatric disorders and HIV/SIV infection. Most evidence indicates that monocytes/macrophages are the primary cell type infected within the CNS and that they contribute to CNS inflammation and neurological disease. Substance P (SP), a pleotropic neuropeptide implicated in inflammation, depression, and immune modulation via interaction with its cognate receptor, the neurokinin 1 receptor (NK1-R), is produced by monocyte/macrophages. While the presence of NK1-R on neurons is well known, its role on cells of the immune system such as monocyte/macrophages is just beginning to emerge. Therefore, we have examined the expression of SP and NK1-R and their relationship to SIV/HIV encephalitis (SIVE/HIVE) lesions and SIV-infected cells. These studies demonstrated intense expression of SP and NK1-R in SIVE lesions, with macrophages being the principal cell expressing NK1-R. Interestingly, all of the SIV-infected macrophages expressed NK1-R. Additionally, we examined the functional role of SP as a proinflammatory mediator of monocyte activation and chemotaxis. These studies demonstrated that treatment of monocytes with SP elicited changes in cell-surface expression for CCR5 and NK1-R in a dose-dependent manner. Moreover, pretreatment with SP enhanced both SP- and CCL5-mediated chemotaxis. All of these findings suggest that SP and NK1-R are important in SIV infection of macrophages and the development of SIVE lesions.
Assuntos
Encéfalo/metabolismo , Receptores da Neurocinina-1/metabolismo , Síndrome de Imunodeficiência Adquirida dos Símios/metabolismo , Vírus da Imunodeficiência Símia/imunologia , Substância P/metabolismo , Animais , Astrócitos/imunologia , Astrócitos/metabolismo , Astrócitos/virologia , Encéfalo/imunologia , Encéfalo/virologia , Movimento Celular , Células Cultivadas , Citometria de Fluxo , Imuno-Histoquímica , Macaca mulatta , Macrófagos/imunologia , Macrófagos/metabolismo , Macrófagos/virologia , Microscopia Confocal , Neurônios/imunologia , Neurônios/metabolismo , Neurônios/virologia , Receptores da Neurocinina-1/imunologia , Síndrome de Imunodeficiência Adquirida dos Símios/imunologia , Síndrome de Imunodeficiência Adquirida dos Símios/virologia , Substância P/imunologiaRESUMO
Neuromediators like substance P have a decisive influence on inflammatory processes via the neuroendocrine regulation circuit. The aim of the present study was therefore to evaluate the expression of the main substance P receptor NK-1R in cattle as well as the modulatory properties of substance P for bovine macrophages. The expression of NK-1R was detected in subsets of lymphocytes, granulocytes, monocytes and in vitro-generated macrophages (MdM). Stimulation of monocytes and MdM with lipopolysaccharide (LPS) for 3h did not alter the expression level of NK-1R. In vitro, the modulatory potential of substance P for monocytes and in vitro-generated blood-derived macrophages (MdM) was analysed. In MdM, generated in the presence of substance P, mRNA expression of chemokines, which are crucial for the attraction and activation of granulocytes and monocytes (CXCL8, CCL5) as well as the expression of IL-1beta, a classically pro-inflammatory cytokine were significantly elevated. After stimulation with LPS, MdM generated in the presence of substance P showed an elevated expression of CXCL8 and IL-1beta, while in SP-influenced monocytes only the expression of CCL5 was significantly upregulated after LPS stimulation. In addition, supernatants of MdM cultured in the presence of substance P induced neutrophil migration and inhibited both necrosis and apoptosis of neutrophil granulocytes. Thus, it has been shown that the modulation of the expression pattern of chemokines and cytokines in MdM by substance P has also functional relevance for the attraction and activation of other immune cells. In general, the modulation of sensor and effector functions by substance P suggests, that this neuromediator can alter the course of an inflammatory disease in cattle.