RESUMO
Neuromodulation of immune function by stimulating the autonomic connections to the spleen has been demonstrated in rodent models. Consequently, neuroimmune modulation has been proposed as a new therapeutic strategy for the treatment of inflammatory conditions. However, demonstration of the translation of these immunomodulatory mechanisms in anatomically and physiologically relevant models is still lacking. Additionally, translational models are required to identify stimulation parameters that can be transferred to clinical applications of bioelectronic medicines. Here, we performed neuroanatomical and functional comparison of the mouse, rat, pig, and human splenic nerve using in vivo and ex vivo preparations. The pig was identified as a more suitable model of the human splenic innervation. Using functional electrophysiology, we developed a clinically relevant marker of splenic nerve engagement through stimulation-dependent reversible reduction in local blood flow. Translation of immunomodulatory mechanisms were then assessed using pig splenocytes and two models of acute inflammation in anesthetized pigs. The pig splenic nerve was shown to locally release noradrenaline upon stimulation, which was able to modulate cytokine production by pig splenocytes. Splenic nerve stimulation was found to promote cardiovascular protection as well as cytokine modulation in a high- and a low-dose lipopolysaccharide model, respectively. Importantly, splenic nerve-induced cytokine modulation was reproduced by stimulating the efferent trunk of the cervical vagus nerve. This work demonstrates that immune responses can be modulated by stimulation of spleen-targeted autonomic nerves in translational species and identifies splenic nerve stimulation parameters and biomarkers that are directly applicable to humans due to anatomical and electrophysiological similarities.
Assuntos
Sistema Imunitário/inervação , Imunomodulação/efeitos dos fármacos , Baço/imunologia , Sistema Nervoso Simpático/imunologia , Nervo Vago/imunologia , Animais , Feminino , Expressão Gênica , Humanos , Sistema Imunitário/efeitos dos fármacos , Inflamação , Interleucina-6/genética , Interleucina-6/imunologia , Lipopolissacarídeos/farmacologia , Camundongos , Microcirculação/efeitos dos fármacos , Microcirculação/genética , Microcirculação/imunologia , Norepinefrina/farmacologia , Ratos , Especificidade da Espécie , Baço/efeitos dos fármacos , Baço/inervação , Baço/patologia , Suínos , Sistema Nervoso Simpático/efeitos dos fármacos , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/imunologia , Nervo Vago/efeitos dos fármacos , Estimulação do Nervo Vago/métodosRESUMO
Endocan is a water-soluble proteoglycan exclusively secreted by vascular endothelium. Endocan levels may be elevated in kidney transplant recipients experiencing antibody-mediated rejection (ABMR), which is characterized by vascular inflammation in transplanted kidney. We evaluated the clinical relevance of endocan as markers of microvascular inflammation in patients who underwent kidney transplantation. Plasma and urinary endocan levels were measured in 203 kidney transplant recipients and were compared across different etiologies of allograft dysfunction and various pathologic scores. Both plasma and urinary endocan levels were significantly higher in patients with acute ABMR than those in patients with normal pathology, acute tubular necrosis (ATN), acute pyelonephritis, BK virus associated nephropathy (BKVN), and T-cell mediated rejection (TCMR). Patients with chronic active ABMR also exhibited significantly higher plasma and urinary endocan levels than patients with long-term graft survival. Scores of glomerulitis and peritubular capillaritis, which are typical features of microvascular inflammation, were significantly elevated in patients with higher plasma and/or urinary endocan levels. Furthermore, plasma and urinary endocan levels could effectively discriminate ABMR from ATN, BKVN, and TCMR. Finally, patients exhibiting high urinary and plasma endocan levels in acute ABMR group showed significantly worse renal survival. Altogether, plasma and urinary endocan levels may serve as potential markers of microvascular inflammation in kidney transplant recipients.
Assuntos
Inflamação/imunologia , Falência Renal Crônica/cirurgia , Transplante de Rim , Microcirculação/imunologia , Proteínas de Neoplasias/sangue , Proteínas de Neoplasias/urina , Proteoglicanas/sangue , Proteoglicanas/urina , Adulto , Área Sob a Curva , Biópsia , Feminino , Rejeição de Enxerto , Humanos , Masculino , Pessoa de Meia-Idade , Necrose , Infecções por Polyomavirus/metabolismo , Pielonefrite/imunologia , Curva ROC , Estudos Retrospectivos , Linfócitos T/citologia , Transplantados , Resultado do Tratamento , Infecções Tumorais por Vírus/imunologiaRESUMO
BACKGROUND: Compared to uninfected adults, HIV-infected adults on antiretroviral therapy are at increased risk of cardiovascular disease. Given the increase in T-cell dysfunction, inflammation, and coagulation in HIV infection, microvascular dysfunction is thought to contribute to this excess cardiovascular risk. However, the relationships between these variables remain undefined. METHODS AND RESULTS: This was a cross-sectional study of 358 HIV-infected adults from the SCOPE cohort. Macrovascular endothelial function was assessed using flow-mediated dilation of the brachial artery and microvascular function by reactive hyperemia. T-cell phenotype was determined by flow cytometry. Plasma markers of inflammation (tumor necrosis factor-α, interleukin-6, high-sensitivity C-reactive protein, sCD14) and coagulation (fibrinogen, D-dimer) were also measured. In all HIV+ subjects, markers of inflammation (tumor necrosis factor-α, high-sensitivity C-reactive protein), coagulation (D-dimer) and T-cell activation (CD8+PD1+, CD4+interferon+cytomegalovirus-specific) were associated with worse reactive hyperemia after adjusting for traditional cardiovascular risk factors and co-infections. In treated and suppressed subjects, tumor necrosis factor-α and CD8+PD1+ cells remained associated with worse reactive hyperemia after adjustment. Compared to the untreated subjects, CD8+PD1+ cells were increased in the virally suppressed group. Reactive hyperemia was predictive of flow-mediated dilation. CONCLUSIONS: CD8+PD1+ cells and tumor necrosis factor-α were associated with microvascular dysfunction in all HIV+ subjects and the treated and suppressed group. Additionally, D-dimer, high-sensitivity C-reactive protein, sCD-14, and interleukin-6 were associated with microvascular dysfunction in all HIV+ subjects. Although T-cell dysfunction, inflammation, and microvascular dysfunction are thought to play a role in cardiovascular disease in HIV, this study is the first to look at which T-cell and inflammatory markers are associated with microvascular dysfunction in HIV-infected individuals.
Assuntos
Coagulação Sanguínea/imunologia , Linfócitos T CD8-Positivos/imunologia , Doenças Cardiovasculares/imunologia , Infecções por HIV/imunologia , Microcirculação/imunologia , Adulto , Artéria Braquial/imunologia , Artéria Braquial/fisiopatologia , Proteína C-Reativa/imunologia , Doenças Cardiovasculares/complicações , Doenças Cardiovasculares/fisiopatologia , Estudos Transversais , Feminino , Produtos de Degradação da Fibrina e do Fibrinogênio/imunologia , Fibrinogênio/imunologia , Infecções por HIV/complicações , Humanos , Hiperemia , Inflamação/imunologia , Interleucina-6/imunologia , Receptores de Lipopolissacarídeos/imunologia , Ativação Linfocitária/imunologia , Masculino , Microvasos , Pessoa de Meia-Idade , Linfócitos T/imunologia , Fator de Necrose Tumoral alfa/imunologia , VasodilataçãoRESUMO
Glucocorticoid-induced leucine zipper (GILZ) is an anti-inflammatory protein first identified in T lymphocytes. We recently observed that GILZ is highly expressed in synovial endothelial cells in rheumatoid arthritis. However, the function of GILZ in endothelial cells is unknown. To investigate the actions of GILZ in this cell type, we induced GILZ expression in HUVECs via transient transfection. GILZ overexpression significantly reduced the capacity of TNF-stimulated HUVECs to support leukocyte rolling, adhesion, and transmigration. These effects were associated with decreased expression of E-selectin, ICAM-1, CCL2, CXCL8, and IL-6. Experiments in a human microvascular endothelial cell line demonstrated that TNF-inducible NF-κB activity was significantly inhibited by overexpression of GILZ. Exogenous GILZ inhibited TNF-induced NF-κB p65 DNA binding, although this occurred in the absence of an effect on p65 nuclear translocation, indicating that the mechanism of action of exogenous GILZ in endothelial cells differs from that reported in other cell types. GILZ overexpression also inhibited TNF-induced activation of p38, ERK, and JNK MAPKs, as well as increased expression of the MAPK inhibitory phosphatase, MKP-1. In contrast, silencing endogenous GILZ in glucocorticoid-treated HUVECs did not alter their capacity to support leukocyte interactions. These data demonstrate that exogenous GILZ exerts inhibitory effects on endothelial cell adhesive function via a novel pathway involving modulation of NF-κB p65 DNA binding and MAPK activity. Induction of GILZ expression in endothelial cells may represent a novel therapeutic modality with the potential to inhibit inflammatory leukocyte recruitment.
Assuntos
Endotélio Vascular/imunologia , Endotélio Vascular/metabolismo , Sistema de Sinalização das MAP Quinases/imunologia , Fator de Transcrição RelA/metabolismo , Fatores de Transcrição/genética , Migração Transendotelial e Transepitelial/imunologia , Adesão Celular/genética , Adesão Celular/imunologia , Comunicação Celular/imunologia , Linhagem Celular , Inibição de Migração Celular/genética , Inibição de Migração Celular/imunologia , Endotélio Vascular/fisiologia , Células Endoteliais da Veia Umbilical Humana , Humanos , Leucócitos/imunologia , Leucócitos/metabolismo , Sistema de Sinalização das MAP Quinases/genética , Microcirculação/genética , Microcirculação/imunologia , Cultura Primária de Células , Distribuição Aleatória , Fatores de Transcrição/biossíntese , Fatores de Transcrição/fisiologia , Migração Transendotelial e Transepitelial/genéticaRESUMO
BACKGROUND: In the setting of inflammatory bowel disease, inflammation is associated with a simultaneous increase in angiogenesis; moreover, elevated vascular endothelial growth factor (VEGF) levels implicate angiogenesis as a pathologic contributor to disease severity. We hypothesize that selectively inhibiting vascular endothelial growth factor receptor-2 (VEGFR2) in a model of murine colitis will reduce angiogenesis, resulting in decreased inflammation and disease severity, providing mechanistic insight into the role of pathologic angiogenesis in IBD. MATERIALS AND METHODS: In a dextran sodium sulfate model of murine colitis, anti-VEGFR2 monoclonal antibody (DC101) or placebo was administered. Clinical assessments followed by histologic and molecular tissue analysis were performed to quantify inflammation, microvessel density (MVD), VEGF and VEGFR2 gene expression, and phosphorylated mitogen-activated protein kinase protein expression. RESULTS: Weight loss began after d 6 with the treatment group demonstrating a more favorable percent weight change. Inflammation and MVD were similar between cohorts, both increasing in parallel toward a plateau. VEGF and VEGFR2 messenger RNA expression were not significantly different, but phosphorylated mitogen-activated protein kinase was elevated in the DC101 cohort (P = 0.03). CONCLUSIONS: Despite a more favorable weight change profile in the treated group, no difference was observed between cohorts regarding clinical disease severity. However, a parallel rise in inflammation and MVD was observed coinciding with weight loss, suggesting their relationship in IBD. VEGFR2 downstream signaling was significantly elevated in the treated cohort, possibly by VEGF-independent signal transduction. Early and effective inhibition of angiogenesis by limiting downstream VEGF signaling or targeting multiple angiogenic pathways may block angiogenesis, thereby reducing disease severity and provide evidence toward the mechanism and clinical benefit of antiangiogenics in the setting of IBD.
Assuntos
Anticorpos Monoclonais/farmacologia , Colite/tratamento farmacológico , Neovascularização Patológica/tratamento farmacológico , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/antagonistas & inibidores , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/imunologia , Doença Aguda , Animais , Peso Corporal/efeitos dos fármacos , Colite/induzido quimicamente , Colite/imunologia , Sulfato de Dextrana/farmacologia , Modelos Animais de Doenças , Expressão Gênica/efeitos dos fármacos , Expressão Gênica/imunologia , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Sistema de Sinalização das MAP Quinases/imunologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Microcirculação/efeitos dos fármacos , Microcirculação/imunologia , Neovascularização Patológica/imunologia , Fator A de Crescimento do Endotélio Vascular/genética , Fator A de Crescimento do Endotélio Vascular/imunologia , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/genéticaRESUMO
Endothelial surface microstructures have been described previously under inflammatory conditions; however, they remain ill-characterized. In this study, CXCL8, an inflammatory chemokine, was shown to induce the formation of filopodia-like protrusions on endothelial cells; the same effects were observed with CXCL10 and CCL5. Chemokines stimulated filopodia formation by both microvascular (from bone marrow and skin) and macrovascular (from human umbilical vein) endothelial cells. Use of blocking Abs and degradative enzymes demonstrated that CXCL8-stimulated filopodia formation was mediated by CXCR1 and CXCR2, Duffy Ag/receptor for chemokines, heparan sulfate (HS), and syndecans. HS was present on filopodial protrusions appearing as a meshwork on the cell surface, which colocalized with CXCL8, and this glycosaminoglycan was 2,6-O- and 3-O-sulfated. Transmission electron microscopy revealed that CXCL8-stimulated filopodial and microvilli-like protrusions that interacted with leukocytes before transendothelial migration and removal of HS reduced this migration. iTRAQ mass spectrometry showed that changes in the levels of cytoskeletal, signaling, and extracellular matrix proteins were associated with CXCL8-stimulated filopodia/microvilli formation; these included tropomyosin, fascin, and Rab7. This study suggests that chemokines stimulate endothelial filopodia and microvilli formation, leading to their presentation to leukocytes and leukocyte transendothelial migration.
Assuntos
Células Apresentadoras de Antígenos/imunologia , Células Apresentadoras de Antígenos/metabolismo , Endotélio Vascular/imunologia , Endotélio Vascular/metabolismo , Interleucina-8/metabolismo , Linhagem Celular Transformada , Linhagem Celular Tumoral , Células Cultivadas , Quimiocina CCL5/metabolismo , Quimiocina CXCL10/metabolismo , Endotélio Vascular/citologia , Células Endoteliais da Veia Umbilical Humana , Humanos , Leucócitos/citologia , Leucócitos/imunologia , Leucócitos/metabolismo , Microcirculação/imunologia , Propriedades de Superfície , Migração Transendotelial e Transepitelial/imunologiaRESUMO
Recent studies suggest dilated Virchow-Robin Spaces (dVRS) could be a manifestation of cerebral small-vessel disease, but little is known about their risk factors. As inflammation has been associated with other brain MRI findings, we investigated whether interleukin-6 and C-reactive protein were associated with the severity of dVRS in the eldery. dVRS were assessed in basal ganglia and white matter and rated on a severity scale. We found that elevated interleukin-6 levels were associated with higher severity of dVRS in basal ganglia, suggesting that inflammation might be associated with the burden of dVRS in the elderly.
Assuntos
Proteína C-Reativa/imunologia , Circulação Cerebrovascular/imunologia , Transtornos Cerebrovasculares/imunologia , Encefalite/imunologia , Interleucina-6/imunologia , Microcirculação/imunologia , Idoso , Envelhecimento/patologia , Gânglios da Base/irrigação sanguínea , Gânglios da Base/patologia , Doenças dos Gânglios da Base/epidemiologia , Doenças dos Gânglios da Base/imunologia , Doenças dos Gânglios da Base/patologia , Proteína C-Reativa/metabolismo , Transtornos Cerebrovasculares/epidemiologia , Transtornos Cerebrovasculares/patologia , Doenças Desmielinizantes/epidemiologia , Doenças Desmielinizantes/imunologia , Doenças Desmielinizantes/patologia , Dilatação Patológica/epidemiologia , Dilatação Patológica/imunologia , Dilatação Patológica/patologia , Encefalite/epidemiologia , Encefalite/patologia , Feminino , Humanos , Interleucina-6/metabolismo , Leucoencefalopatias/epidemiologia , Leucoencefalopatias/imunologia , Leucoencefalopatias/patologia , Imageamento por Ressonância Magnética , Masculino , Fatores de Risco , Índice de Gravidade de DoençaRESUMO
During neuroinflammation, cytokines such as TNF-α and IFN-γ secreted by activated leukocytes and/or CNS resident cells have been shown to alter the phenotype and function of brain endothelial cells (BECs) leading to blood-brain barrier breakdown. In this study, we show that the human BEC line hCMEC/D3 expresses the receptors for TNF-α, TNF receptor 1 and TNF receptor 2, and for IFN-γ. BEC activation with TNF-α alone or in combination with IFN-γ induced endothelial leakage of paracellular tracers. At high cytokine concentrations (10 and 100 ng/ml), this effect was associated with caspase-3/7 activation and apoptotic cell death as evidenced by annexin V staining and DNA fragmentation (TUNEL) assays. In addition, inhibition of JNK and protein kinase C activation at these doses partially prevented activation of caspase-3/7, although only JNK inhibition was partially able to prevent the increase in BEC paracellular permeability induced by cytokines. By contrast, lower cytokine concentrations (1 ng/ml) also led to effector caspase activation, increased paracellular flux, and redistribution of zonula occludens-1 and VE-cadherin but failed to induce apoptosis. Under these conditions, specific caspase-3 and caspase-9, but not caspase-8, inhibitors partially blocked cytokine-induced disruption of tight and adherens junctions and BEC paracellular permeability. Our results suggest that the concentration of cytokines in the CNS endothelial microenvironment determines the extent of caspase-mediated barrier permeability changes, which may be generalized as a result of apoptosis or more subtle as a result of alterations in the organization of junctional complex molecules.
Assuntos
Barreira Hematoencefálica/enzimologia , Barreira Hematoencefálica/imunologia , Encéfalo/enzimologia , Encéfalo/imunologia , Citocinas/fisiologia , Endotélio Vascular/enzimologia , Endotélio Vascular/imunologia , Barreira Hematoencefálica/patologia , Encéfalo/patologia , Linhagem Celular , Endotélio Vascular/patologia , Humanos , Mediadores da Inflamação/metabolismo , Mediadores da Inflamação/fisiologia , Interferon gama/metabolismo , Microcirculação/imunologia , Receptores de Interferon/biossíntese , Receptores Tipo I de Fatores de Necrose Tumoral/biossíntese , Receptores Tipo I de Fatores de Necrose Tumoral/metabolismo , Receptores Tipo II do Fator de Necrose Tumoral/biossíntese , Receptores Tipo II do Fator de Necrose Tumoral/metabolismo , Frações Subcelulares/enzimologia , Frações Subcelulares/imunologia , Frações Subcelulares/patologia , Receptor de Interferon gamaRESUMO
Mononuclear cell migration into the vascular subendothelium constitutes an early event of the atherogenic process. Because the effect of retinoid X receptor (RXR)α on arterial mononuclear leukocyte recruitment is poorly understood, this study investigated whether RXR agonists can affect this response and the underlying mechanisms involved. Decreased RXRα expression was detected after 4 h stimulation of human umbilical arterial endothelial cells with TNF-α. Interestingly, under physiological flow conditions, TNF-α-induced endothelial adhesion of human mononuclear cells was concentration-dependently inhibited by preincubation of the human umbilical arterial endothelial cells with RXR agonists such as bexarotene or 9-cis-retinoid acid. RXR agonists also prevented TNF-α-induced VCAM-1 and ICAM-1 expression, as well as endothelial growth-related oncogene-α and MCP-1 release. Suppression of RXRα expression with a small interfering RNA abrogated these responses. Furthermore, inhibition of MAPKs and NF-κB pathways were involved in these events. RXR agonist-induced antileukocyte adhesive effects seemed to be mediated via RXRα/peroxisome proliferator-activated receptor (PPAR)γ interaction, since endothelial PPARγ silencing abolished their inhibitory responses. Furthermore, RXR agonists increased RXR/PPARγ interaction, and combinations of suboptimal concentrations of both nuclear receptor ligands inhibited TNF-α-induced mononuclear leukocyte arrest by 60-65%. In vivo, bexarotene dose-dependently inhibited TNF-α-induced leukocyte adhesion to the murine cremasteric arterioles and decreased VCAM-1 and ICAM-1 expression. Therefore, these results reveal that RXR agonists can inhibit the initial inflammatory response that precedes the atherogenic process by targeting different steps of the mononuclear recruitment cascade. Thus, RXR agonists may constitute a new therapeutic tool in the control of the inflammatory process associated with cardiovascular disease.
Assuntos
Inibição de Migração Celular/imunologia , Endotélio Vascular/imunologia , Endotélio Vascular/patologia , Leucócitos Mononucleares/imunologia , Leucócitos Mononucleares/patologia , PPAR gama/metabolismo , Receptor X Retinoide alfa/agonistas , Bexaroteno , Comunicação Celular/efeitos dos fármacos , Comunicação Celular/imunologia , Linhagem Celular , Inibição de Migração Celular/efeitos dos fármacos , Regulação para Baixo/efeitos dos fármacos , Regulação para Baixo/imunologia , Endotélio Vascular/metabolismo , Humanos , Molécula 1 de Adesão Intercelular/sangue , Leucócitos Mononucleares/efeitos dos fármacos , Microcirculação/efeitos dos fármacos , Microcirculação/imunologia , Músculo Esquelético/irrigação sanguínea , Músculo Esquelético/efeitos dos fármacos , Músculo Esquelético/imunologia , PPAR gama/fisiologia , Receptor X Retinoide alfa/biossíntese , Receptor X Retinoide alfa/fisiologia , Tetra-Hidronaftalenos/farmacologia , Fator de Necrose Tumoral alfa/antagonistas & inibidores , Fator de Necrose Tumoral alfa/fisiologia , Artérias Umbilicais/efeitos dos fármacos , Artérias Umbilicais/imunologia , Artérias Umbilicais/patologia , Molécula 1 de Adesão de Célula Vascular/sangueAssuntos
Síndrome Antifosfolipídica/diagnóstico , Hemorragia/diagnóstico , Programas de Rastreamento/métodos , Angioscopia Microscópica/métodos , Trombose/diagnóstico , Adulto , Síndrome Antifosfolipídica/imunologia , Feminino , Hemorragia/imunologia , Humanos , Masculino , Microcirculação/imunologia , Pessoa de Meia-Idade , Trombose/imunologiaRESUMO
During Gram-negative sepsis and endotoxemia, CD14 is essential for the recognition of LPS by the TLR4 complex and subsequent generation of systemic inflammation. However, CD14-independent responses to LPS have been reported in vitro and in vivo in selected tissues including the skin. As the liver is a key target organ for neutrophil sequestration and inflammatory pathology during sepsis and endotoxemia, we investigated the role of CD14 in the recruitment of neutrophils into the liver in a mouse model of endotoxemia. Using dynamic in vivo imaging of the liver, we observed that neutrophil recruitment within the sinusoids and post-sinusoidal venules occurred equivalently between LPS-treated wild-type and CD14-knockout mice. Neutrophil recruitment within the liver was completely independent of CD14 regardless of whether it was expressed on cells of hematopoietic or nonhematopoietic origin or in serum as soluble CD14. Whereas CD14 expression was essential for activation of circulating neutrophils and for the development of LPS-induced systemic inflammation (pulmonary neutrophil sequestration, leukopenia, and increased serum proinflammatory cytokine levels), deficiency of CD14 did not limit the adhesion strength of neutrophils in vitro. Furthermore, wild-type and CD14-knockout mice displayed identical deposition of serum-derived hyaluronan-associated protein within liver sinusoids in response to LPS, indicating that the sinusoid-specific CD44/hyaluronan/serum-derived hyaluronan-associated protein-dependent pathway of neutrophil adhesion is activated independently of CD14. Therefore, the liver microcirculation possesses a unique CD14-independent mechanism of LPS detection and activation of neutrophil recruitment.
Assuntos
Endotoxemia/imunologia , Endotoxemia/patologia , Receptores de Lipopolissacarídeos/fisiologia , Circulação Hepática/imunologia , Microcirculação/imunologia , Infiltração de Neutrófilos/imunologia , Animais , Receptor 1 de Quimiocina CX3C , Modelos Animais de Doenças , Endotélio Vascular/imunologia , Endotélio Vascular/metabolismo , Endotélio Vascular/patologia , Endotoxemia/sangue , Feminino , Células-Tronco Hematopoéticas/imunologia , Células-Tronco Hematopoéticas/metabolismo , Células-Tronco Hematopoéticas/patologia , Imunidade Inata/genética , Mediadores da Inflamação/fisiologia , Receptores de Lipopolissacarídeos/sangue , Receptores de Lipopolissacarídeos/genética , Lipopolissacarídeos/administração & dosagem , Circulação Hepática/genética , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Microcirculação/genética , Infiltração de Neutrófilos/genética , Receptores de Quimiocinas/genética , Transdução de Sinais/genética , Transdução de Sinais/imunologiaRESUMO
The significance of C4d-Banff scores in protocol biopsies of kidney transplant recipients with preformed donor-specific antibodies (DSA) has not been determined. We reviewed 157 protocol biopsies from 80 DSA+ patients obtained at 3 months and 1 year post-transplant. The C4d Banff scores (1,2,3) were associated with significant increments of microcirculation inflammation (MI) at both 3 months and 1 year post-transplant, worse transplant glomerulopathy and higher class II DSA-MFI (p < 0.01). Minimal-C4d had injury intermediate between negative and focal, while focal and diffuse-C4d had the same degree of microvascular injury. A total of 54% of patients had variation of C4d score between 3 months and 1 year post-transplant. Cumulative (3 month + 1 year) C4d scores correlated with long-term renal function worsening (p = 0.006). However, C4d staining was not a sensitive indicator of parenchymal disease, 55% of C4d-negative biopsies having evidence of concomitant MI. Multivariate analysis demonstrated that the presence of MI and class II DSA at 3 months were associated with a fourfold increased risk of progression to chronic antibody-mediated rejection independently of C4d (p < 0.05). In conclusion, the substantial fluctuation of C4d status in the first year post-transplant reflects a dynamic humoral process. However, C4d may not be a sufficiently sensitive indicator of activity, MI and DSA being more robust predictors of bad outcome.
Assuntos
Complemento C4b/imunologia , Rejeição de Enxerto/imunologia , Transplante de Rim/imunologia , Fragmentos de Peptídeos/imunologia , Adulto , Idoso , Anticorpos , Biópsia , Sobrevivência de Enxerto/imunologia , Humanos , Rim/imunologia , Transplante de Rim/patologia , Microcirculação/imunologia , Pessoa de Meia-IdadeRESUMO
INTRODUCTION: Lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1), the major endothelial receptor for oxidized low-density lipoprotein, is also involved in leukocyte recruitment. Systemic leukocyte activation in sepsis represents a crucial factor in the impairment of the microcirculation of different tissues, causing multiple organ failure and subsequently death. The aim of our experimental study was to evaluate the effects of LOX-1 inhibition on the endotoxin-induced leukocyte adherence and capillary perfusion within the intestinal microcirculation by using intravital microscopy (IVM). METHODS: We used 40 male Lewis rats for the experiments. Ten placebo-treated animals served as a control. Thirty animals received 5 mg/kg lipopolysaccharide (LPS) intravenously. Ten endotoxemic rats remained untreated. In 10 LPS animals, we administered additionally 10 mg/kg LOX-1 antibodies. Ten further LPS animals received a nonspecific immunoglobulin (rat IgG) intravenously. After 2 hours of observation, intestinal microcirculation was evaluated by using IVM; the plasma levels of monocyte chemoattractant protein-1 (MCP-1) and tumor necrosis factor-alpha (TNF-α) were determined; and LOX-1 expression was quantified in intestinal tissue with Western blot and reverse-transcription polymerase chain reaction (PCR). RESULTS: LOX-1 inhibition significantly reduced LPS-induced leukocyte adhesion in intestinal submucosal venules (P < 0.05). At the protein and mRNA levels, LOX-1 expression was significantly increased in untreated LPS animals (P < 0.05), whereas in animals treated with LOX-1 antibody, expression of LOX-1 was reduced (P < 0.05). MCP-1 plasma level was reduced after LOX-1 antibody administration. CONCLUSIONS: Inhibition of LOX-1 reduced leukocyte activation in experimental endotoxemia. LOX-1 represents a novel target for the modulation of the inflammatory response within the microcirculation in sepsis.
Assuntos
Anticorpos Antibacterianos/uso terapêutico , Endotoxemia/patologia , Mucosa Intestinal/irrigação sanguínea , Mucosa Intestinal/patologia , Leucócitos/imunologia , Microcirculação/imunologia , Receptores Depuradores Classe E/antagonistas & inibidores , Animais , Anticorpos Antibacterianos/farmacologia , Adesão Celular/imunologia , Endotoxemia/imunologia , Mucosa Intestinal/imunologia , Leucócitos/patologia , Masculino , Ratos , Ratos Endogâmicos Lew , Receptores Depuradores Classe E/imunologiaRESUMO
PURPOSE OF REVIEW: The purpose of this review is to summarize the recent experimental and clinical literature on the pathogenesis of transfusion-related acute lung injury (TRALI). RECENT FINDINGS: In both experimental and clinical TRALI, an immune priming step is generally necessary to produce lung injury. Experimental studies have used mainly lipopolysaccharide (LPS) as the priming step, whereas in clinical TRALI the specific priming events are currently being defined and include recent surgery and active infections. Experimental studies have modeled TRALI by using anti-major histocompatibility complex antibodies, antineutrophil antibodies, and also bioactive lipids isolated from stored human blood. A common theme among the experimental TRALI models is the central importance of neutrophils in mediating the early immune response and lung vascular injury. New work has focused on the interplay between neutrophils and platelets in the lung microcirculation. Finally, plasma mitigation strategies implemented in several countries are showing early promise in decreasing the incidence of TRALI from high plasma volume blood products. SUMMARY: TRALI requires an immune priming step followed by transfusion of a blood product with either leukocyte allo-antibodies or biological response modifiers. TRALI invokes an acute immune response dominated by neutrophils interacting with platelets and the lung endothelium.
Assuntos
Lesão Pulmonar Aguda/imunologia , Lesão Pulmonar Aguda/fisiopatologia , Reação Transfusional , Lesão Pulmonar Aguda/etiologia , Plaquetas/imunologia , Endotélio Vascular , Humanos , Isoanticorpos , Pulmão/irrigação sanguínea , Pulmão/imunologia , Microcirculação/imunologia , Neutrófilos/imunologiaRESUMO
Much information can be gained by investigating the consequences of hyperthermia on individual cell populations in vitro, however the precise effects of such a therapeutic modality in vivo depend on the tumour microenvironment and the cellular composition therein. Although the direct cytotoxic effects of hyperthermia on tumour tissue can lead to an immediate reduction in tumour volume, long-term benefits to local and distal tumour recurrence will very much depend on the induction of immunity and the capacity of effector cells to traffic to tumours and elicit their cytotoxic functions. The immunological sequelae to hyperthermia are even more important in those instances when large tumour volumes preclude the delivery of appropriate thermal damage. The development of protective anti-tumour immunity requires a plethora of interactions and responses, the vast majority of which can be influenced by temperatures that are consistent with fever-like temperatures (39 degrees -40 degrees C), as well as hyperthermia treatment (<41 degrees C). This article reviews current knowledge relating to the effects of hyperthermia treatment on aspects of the induction and manifestation of immunological responses that are most pertinent to the development and maintenance of protective anti-tumour immunity.
Assuntos
Citotoxicidade Imunológica/imunologia , Hipertermia Induzida , Neoplasias/imunologia , Neoplasias/terapia , Células Apresentadoras de Antígenos/imunologia , Humanos , Terapia de Imunossupressão , Microcirculação/imunologia , Neoplasias/irrigação sanguínea , Linfócitos T/imunologiaRESUMO
OBJECTIVE: During abdominal sepsis, the activation of hepatic Kupffer cells (KC) and its consequences are of central interest. This study evaluates the impact of selective KC depletion on hepatic microcirculation, cytokine release, and systemic alterations in the colon ascendens stent peritonitis (CASP), a model of polymicrobial abdominal sepsis. METHODS: For KC depletion clodronate liposomes were injected 24 h before CASP surgery in female C57BL/6N mice. Three and 12 h after CASP, in-vivo fluorescence microscopy of the liver was performed. Analysis of hepatocellular apoptosis was conducted by immunohistochemistry. In addition, levels of tumor necrosis factor (TNF), IL-6, and IL-10 in the liver, lungs, spleen, and plasma were determined, and bacteriology and survival analysis were performed. RESULTS: CASP led to significant sinusoidal perfusion failure, increased leukocyte recruitment, hepatocellular apoptosis and increased levels of TNF, IL-6, and IL-10 in the liver and plasma. KC depletion before CASP significantly reduced leukocyte recruitment to the liver and hepatocellular apoptosis. IL-10 secretion decreased dramatically in the liver and plasma of KC-depleted septic mice. In contrast, TNF levels were clearly elevated after clodronate treatment. In the lung and spleen, a compensatory upregulation of IL-10 could be detected after KC depletion. Clodronate treatment resulted in a significant reduction in survival. CONCLUSION: The results indicate that KC depletion is locally protective in polymicrobial abdominal sepsis, as it reduces hepatic inflammation and apoptosis. These effects could be observed in the presence of clearly elevated TNF levels. However, the lack of IL-10 in KC-depleted mice resulted in a detrimental systemic proinflammation.
Assuntos
Ácido Clodrônico/farmacologia , Hepatite , Células de Kupffer/efeitos dos fármacos , Células de Kupffer/patologia , Lipossomos/farmacologia , Sepse , Animais , Apoptose/efeitos dos fármacos , Apoptose/imunologia , Colo , Citocinas/sangue , Modelos Animais de Doenças , Feminino , Hepatite/imunologia , Hepatite/patologia , Hepatite/terapia , Fígado/irrigação sanguínea , Fígado/imunologia , Fígado/patologia , Camundongos , Camundongos Endogâmicos C57BL , Microcirculação/imunologia , Peritonite/imunologia , Peritonite/patologia , Peritonite/terapia , Sepse/imunologia , Sepse/patologia , Sepse/terapia , Stents , Taxa de SobrevidaRESUMO
BACKGROUND AND PURPOSE: Over three decades of research evaluating the biology of melanocortin (MC) hormones and synthetic peptides, activation of the MC type 1 (MC(1)) receptor has been identified as a viable target for the development of novel anti-inflammatory therapeutic agents. Here, we have tested a recently described selective agonist of MC(1) receptors, BMS-470539, on leucocyte/post-capillary venule interactions in murine microvascular beds. EXPERIMENTAL APPROACH: Intravital microscopy of two murine microcirculations were utilized, applying two distinct modes of promoting inflammation. The specificity of the effects of BMS-470539 was assessed using mice bearing mutant inactive MC(1) receptors (the recessive yellow e/e colony). KEY RESULTS: BMS-470539, given before an ischaemia-reperfusion protocol, inhibited cell adhesion and emigration with no effect on cell rolling, as assessed 90 min into the reperfusion phase. These properties were paralleled by inhibition of tissue expression of both CXCL1 and CCL2. Confocal investigations of inflamed post-capillary venules revealed immunostaining for MC(1) receptors on adherent and emigrated leucocytes. Congruently, the anti-inflammatory properties of BMS-470539 were lost in mesenteries of mice bearing the inactive mutant MC(1) receptors. Therapeutic administration of BMS-470539 stopped cell emigration, but did not affect cell adhesion in the cremasteric microcirculation inflamed by superfusion with platelet-activating factor. CONCLUSIONS AND IMPLICATIONS: Activation of MC(1) receptors inhibited leucocyte adhesion and emigration. Development of new chemical entities directed at MC(1) receptors could be a viable approach in the development of novel anti-inflammatory therapeutic agents with potential application to post-ischaemic conditions.
Assuntos
Quimiotaxia de Leucócito/efeitos dos fármacos , Imidazóis/farmacologia , Microvasos/efeitos dos fármacos , Receptor Tipo 1 de Melanocortina/agonistas , Animais , Quimiocina CCL2/antagonistas & inibidores , Quimiocina CCL2/biossíntese , Quimiocina CXCL1/antagonistas & inibidores , Quimiocina CXCL1/biossíntese , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Microcirculação/efeitos dos fármacos , Microcirculação/imunologia , Microvasos/imunologia , Microvasos/metabolismo , Fator de Ativação de Plaquetas/farmacologia , Receptor Tipo 1 de Melanocortina/genética , Circulação Esplâncnica , Vasculite/imunologia , Vasculite/metabolismoRESUMO
Banff classification empirically established scoring of histologic lesions, but the relationships of lesions to each other and to underlying biologic processes remain unclear. We hypothesized that class discovery tools would reveal new relationships between individual lesions, and relate lesions to C4d staining, anti-HLA donor-specific antibody (DSA) and time posttransplant. We studied 234 nonselected renal allograft biopsies for clinical indications from 173 patients. Silhouette plotting and principal component analysis revealed three groups of lesions: microcirculation changes, including inflammation (glomerulitis, capillaritis) and deterioration (double contours, mesangial expansion); scarring/hyalinosis; and tubulointerstitial inflammation. DSA and C4d grouped with microcirculation inflammation, whereas time posttransplant grouped with scarring/hyalinosis lesions. Intimal arteritis clustered with DSA, C4d and microcirculation inflammation, but also showed correlations with tubulitis. Fibrous intimal thickening in arteries clustered with scarring/hyalinosis. Capillary basement membrane multilayering showed intermediary relationships between microcirculation deterioration and time-dependent scarring. Correlation analysis and hierarchical clustering confirmed the lesion relationships. Thus, we propose that the pathologic lesions in biopsies are not independent but are members of groups that represent distinct pathogenic forces: microcirculation changes, reflecting the stress of DSA; scarring, hyalinosis and arterial fibrosis, reflecting the cumulative burden of injury over time; and tubulointerstitial inflammation. Interpretation of lesions should reflect these associations.
Assuntos
Transplante de Rim/imunologia , Rim/patologia , Arterite/imunologia , Arterite/patologia , Biópsia , Capilares/imunologia , Capilares/patologia , Cicatriz/imunologia , Cicatriz/patologia , Complemento C4b , Fibrose/patologia , Humanos , Inflamação/imunologia , Inflamação/patologia , Rim/imunologia , Transplante de Rim/patologia , Microcirculação/imunologia , Fragmentos de Peptídeos , Doadores de Tecidos , Vasculite/imunologia , Vasculite/patologiaRESUMO
We studied the phenotype of late kidney graft failure in a prospective study of unselected kidney transplant biopsies taken for clinical indications. We analyzed histopathology, HLA antibodies and death-censored graft survival in 234 consecutive biopsies from 173 patients, taken 6 days to 31 years posttransplant. Patients with late biopsies (>1 year) frequently displayed donor-specific HLA antibody (particularly class II) and microcirculation changes, including glomerulitis, glomerulopathy, capillaritis, capillary multilayering and C4d staining. Grafts biopsied early rarely failed (1/68), whereas grafts biopsied late often progressed to failure (27/105) within 3 years. T-cell-mediated rejection and its lesions were not associated with an increased risk of failure after biopsy. In multivariable analysis, graft failure correlated with microcirculation inflammation and scarring, but C4d staining was not significant. When microcirculation changes and HLA antibody were used to define antibody-mediated rejection, 17/27 (63%) of late kidney failures after biopsy were attributable to antibody-mediated rejection, but many were C4d negative and missed by current diagnostic criteria. Glomerulonephritis accounted for 6/27 late losses, whereas T-cell-mediated rejection, drug toxicity and unexplained scarring were uncommon. The major cause of late kidney transplant failure is antibody-mediated microcirculation injury, but detection of this phenotype requires new diagnostic criteria.
Assuntos
Autoanticorpos/imunologia , Função Retardada do Enxerto/imunologia , Antígenos de Histocompatibilidade Classe II/imunologia , Transplante de Rim/imunologia , Autoanticorpos/sangue , Biópsia , Complemento C4b/imunologia , Função Retardada do Enxerto/epidemiologia , Função Retardada do Enxerto/patologia , Feminino , Glomerulonefrite/epidemiologia , Glomerulonefrite/imunologia , Glomerulonefrite/patologia , Sobrevivência de Enxerto/imunologia , Antígenos de Histocompatibilidade Classe I/imunologia , Humanos , Masculino , Microcirculação/imunologia , Análise Multivariada , Fragmentos de Peptídeos/imunologia , Valor Preditivo dos Testes , Circulação Renal/imunologia , Fatores de Risco , Linfócitos T/imunologiaRESUMO
We studied whether de novo donor-specific antibodies (DSA) in sera from patients undergoing kidney transplant biopsies associate with specific histologic lesions in the biopsy and prognosis. DSA were assessed in 145 patients at the time of biopsy between 7 days to 31 years posttransplant. DSA was detected in 54 patients (37%), of which 32 represented de novo DSA. De novo DSA was more frequent in patients having late biopsies (34%) versus early biopsies (4%), and was usually either against class II alone or class I and II but rarely against class I alone. Microcirculation inflammation (glomerulitis, capillaritis) and damage (glomuerulopathy, capillary basement membrane multilayering), and C4d staining were associated with de novo DSA. However, the degree of scarring, arterial fibrosis and tubulo-interstitial inflammation did not correlate with the presence of de novo DSA. De novo DSA correlated with reduced graft survival after the biopsy. Thus, de novo DSA at the time of a late biopsy for clinical indication is primarily against class II, and associates with microcirculation changes in the biopsy and subsequent graft failure. We propose careful assessment of de novo DSA, particularly against class II, be performed in all late kidney transplant biopsies.