RESUMEN
The purpose of this study was to explore the role and mechanism of transient receptor potential M(2) (TRPM(2)) in antigen-induced arthritis (AIA) mice. Twelve C57BL/6 mice and 12 TRPM(2) knockout mice were divided into 4 groups, includingwild type control group, wild type AIA group, TRPM(2) knockout control group and TRPM(2) knockout AIA group, with 6 mice in each group. Methylated bovine serum albumin (mBSA) was used to establish AIA mouse model. The degree of joint swelling and inflammatory cell infiltration were recorded, as well as synovial hyperplasia of the knee joints. Real-time quantitative polymerase chain reaction (PCR) was used to detect the expression of interleukin (IL)-6, IL-8, chemokine ligand 6 (CXCL-6) and tumor necrosis factor alpha (TNFα) mRNA in synovial cells of knee joints. The results showed that compared with the wild-type AIA group, the TRPM(2) knockout AIA group had more significant synovial proliferation and inflammatory cell infiltration in the synovial tissue.The neutrophil and macrophage counts rather than monocytes in the knee joints of TRPM(2) knockout AIA group were higher than those in wild-type AIA mice. The expression of IL-6, IL-8 and CXCL-6 mRNA were significantly increased in the knock out mice. In summary, TRPM(2) may inhibit inflammatory cytokines such as IL-6 and IL-8 in knee joints of AIA mice by reducing the infiltration of neutrophils and macrophages, the refore alleviates the manifestations of knee arthritis.
Asunto(s)
Artritis Experimental/inmunología , Artritis Reumatoide/inmunología , Quimiocina CXCL6/inmunología , Interleucina-6/inmunología , Interleucina-8/inmunología , Albúmina Sérica Bovina/inmunología , Factor de Necrosis Tumoral alfa/inmunología , Animales , Antígenos/inmunología , Artritis Reumatoide/genética , Quimiocina CXCL6/genética , Interleucina-6/genética , Interleucina-8/genética , Articulación de la Rodilla/inmunología , Articulación de la Rodilla/patología , Ratones , Ratones Endogámicos C57BL , Reacción en Cadena en Tiempo Real de la Polimerasa , Membrana Sinovial , Factor de Necrosis Tumoral alfa/genéticaRESUMEN
Chemokines are a superfamily of structurally related chemotactic cytokines exerting significant roles in regulating cell migration and activation. Currently, five subgroups of fish specific CXC chemokines, named CXCL_F1-CXCL_F5, have been identified in teleost fish. However, understanding of the functions of these fish specific CXC chemokines is still limited. Here, a new member of fish specific CXC chemokines, LcCXCL_F6, was cloned from large yellow croaker Larimichthys crocea. Its open reading frame (ORF) is 369 nucleotides long, encoding a peptide of 122 amino acids (aa). The deduced LcCXCL_F6 protein contains a 19-aa signal peptide and a 103-aa mature polypeptide, which has four conserved cysteine residues (C28, C30, C56, and C72), as found in other known CXC chemokines. Phylogenetic analysis showed LcCXCL_F6 formed a separate clade with sequences from other fish species, tentatively named CXCL_F6, distinct from the clades formed by fish CXCL_F1-5 and mammalian CXC chemokines. The LcCXCL_F6 transcripts were constitutively expressed in all examined tissues and significantly up-regulated in the spleen and head kidney tissues by poly (I:C) and Vibrio alginolyticus. Its transcripts were also detected in primary head kidney leukocytes (HKLs), peripheral blood leucocytes (PBLs), and large yellow croaker head kidney (LYCK) cell line, and significantly up-regulated by poly(I:C), lipopolysaccharide (LPS), and peptidoglycan (PGN) in HKLs. Recombinant LcCXCL_F6 protein (rLcCXCL_F6) could not only chemotactically attract monocytes/macrophages and lymphocytes from PBLs, but also enhance NO release and expression of proinflammatory cytokines (TNF-α, IL-1ß, and CXCL8) in monocytes/macrophages. These results indicate that LcCXCL_F6 plays a role in mediating the inflammatory response.
Asunto(s)
Quimiocina CXCL6/genética , Quimiocina CXCL6/inmunología , Enfermedades de los Peces/inmunología , Regulación de la Expresión Génica/inmunología , Inmunidad Innata/genética , Perciformes/genética , Perciformes/inmunología , Secuencia de Aminoácidos , Animales , Quimiocina CXCL6/química , Proteínas de Peces/química , Proteínas de Peces/genética , Proteínas de Peces/inmunología , Perfilación de la Expresión Génica/veterinaria , Filogenia , Poli I-C/farmacología , Alineación de Secuencia/veterinaria , Vibriosis/inmunología , Vibrio alginolyticus/fisiologíaRESUMEN
Soluble C-X-C chemokine ligand 16 (CXCL16) is related to the inflammatory response in liver injury and involved in the pathogenesis of renal dysfunction in diabetes patients. However, the exact role of elevated CXCL16 in diabetic nephropathy (DN) remains unclear. In this study, we investigated the role of CXCL16 in streptozcin (STZ)-induced diabetic nephropathy (DN) in mice. The results showed that fasting blood glucose (FBG) and 24 h urinary protein, triglyceride, and cholesterol levels increased in diabetic mice, and these changes were partially ameliorated in CXCL16 KO mice. Meanwhile, the results also showed that ROS generation was suppressed and the expression levels of inflammatory factors and infiltration factors were inhibited both in vivo and in vitro using DN models. In addition, the total AKT protein and p-AKT levels were decreased in CXCL16-depleted HK-2 cells that were treated with LPS. These findings suggest that the CXCL16 gene product promotes inflammatory factors and cell infiltration factors, and inhibits the expression of antioxidant factors to accelerate the development of DN, and CXCL16 deficiency attenuates DN may be involved in the AKT signaling pathway.
Asunto(s)
Glucemia/inmunología , Quimiocina CXCL6/inmunología , Nefropatías Diabéticas/inmunología , Inflamación/inmunología , Estrés Oxidativo/inmunología , Especies Reactivas de Oxígeno/inmunología , Animales , Quimiocina CXCL16 , Nefropatías Diabéticas/inducido químicamente , Nefropatías Diabéticas/patología , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , EstreptozocinaRESUMEN
Decidual spiral arteriole (SpA) remodeling is essential to ensure optimal uteroplacental blood flow during human pregnancy, yet very little is known about the regulatory mechanisms. Uterine decidual NK (dNK) cells and macrophages infiltrate the SpAs and are proposed to initiate remodeling before colonization by extravillous trophoblasts (EVTs); however, the trigger for their infiltration is unknown. Using human first trimester placenta, decidua, primary dNK cells, and macrophages, we tested the hypothesis that EVTs activate SpA endothelial cells to secrete chemokines that have the potential to recruit maternal immune cells into SpAs. Gene array, real-time PCR, and ELISA analyses showed that treatment of endothelial cells with EVT conditioned medium significantly increased production of two chemokines, CCL14 and CXCL6. CCL14 induced chemotaxis of both dNK cells and decidual macrophages, whereas CXCL6 also induced dNK cell migration. Analysis of the decidua basalis from early pregnancy demonstrated expression of CCL14 and CXCL6 by endothelial cells in remodeling SpAs, and their cognate receptors are present in both dNK cells and macrophages. Neutralization studies identified IL-6 and CXCL8 as factors secreted by EVTs that induce endothelial cell CCL14 and CXCL6 expression. This study has identified intricate crosstalk between EVTs, SpA cells, and decidual immune cells that governs their recruitment to SpAs in the early stages of remodeling and has identified potential key candidate factors involved. This provides a new understanding of the interactions between maternal and fetal cells during early placentation and highlights novel avenues for research to understand defective SpA remodeling and consequent pregnancy pathology.
Asunto(s)
Arteriolas/fisiología , Decidua/fisiología , Células Endoteliales/metabolismo , Células Asesinas Naturales/fisiología , Macrófagos/fisiología , Trofoblastos/metabolismo , Arteriolas/citología , Arteriolas/inmunología , Movimiento Celular/inmunología , Células Cultivadas , Quimiocina CXCL6/biosíntesis , Quimiocina CXCL6/inmunología , Quimiocinas CC/biosíntesis , Quimiocinas CC/inmunología , Medios de Cultivo/química , Decidua/inmunología , Células Endoteliales/efectos de los fármacos , Células Endoteliales/inmunología , Femenino , Humanos , Interleucina-6/inmunología , Interleucina-6/metabolismo , Interleucina-8/inmunología , Interleucina-8/metabolismo , Macrófagos/inmunología , Placenta/citología , Placenta/inmunología , Embarazo , Primer Trimestre del Embarazo , Trofoblastos/inmunologíaRESUMEN
Recent studies have shown that inflammation plays a critical role in the initiation and progression of hypertensive kidney disease, including renal artery stenosis. However, the signaling mechanisms underlying the induction of inflammation are poorly understood. We found that CXCL16 was induced in the kidney in a murine model of renal artery stenosis. To determine whether CXCL16 is involved in renal injury and fibrosis, wild-type and CXCL16 knockout mice were subjected to renal artery stenosis induced by placing a cuff on the left renal artery. Wild-type and CXCL16 knockout mice had comparable blood pressure at baseline. Renal artery stenosis caused an increase in blood pressure that was similar between wild-type and CXCL16 knockout mice. CXCL16 knockout mice were protected from RAS-induced renal injury and fibrosis. CXCL16 deficiency suppressed bone marrow-derived fibroblast accumulation and myofibroblast formation in the stenotic kidneys, which was associated with less expression of extracellular matrix proteins. Furthermore, CXCL16 deficiency inhibited infiltration of F4/80(+) macrophages and CD3(+) T cells in the stenotic kidneys compared with those of wild-type mice. Taken together, our results indicate that CXCL16 plays a pivotal role in the pathogenesis of renal artery stenosis-induced renal injury and fibrosis through regulation of bone marrow-derived fibroblast accumulation and macrophage and T-cell infiltration.
Asunto(s)
Lesión Renal Aguda/genética , Quimiocina CXCL6/genética , Fibroblastos , Riñón/patología , Macrófagos/inmunología , Obstrucción de la Arteria Renal/genética , Linfocitos T/inmunología , Lesión Renal Aguda/inmunología , Animales , Presión Sanguínea , Western Blotting , Células de la Médula Ósea , Quimiocina CXCL16 , Quimiocina CXCL6/inmunología , Modelos Animales de Enfermedad , Fibrosis/genética , Fibrosis/inmunología , Técnica del Anticuerpo Fluorescente , Frecuencia Cardíaca , Hipertensión/complicaciones , Hipertensión/inmunología , Riñón/inmunología , Riñón/metabolismo , Masculino , Ratones , Ratones Noqueados , Reacción en Cadena en Tiempo Real de la Polimerasa , Obstrucción de la Arteria Renal/inmunología , Insuficiencia Renal Crónica/etiología , Insuficiencia Renal Crónica/inmunología , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
Conventional dendritic cells (cDCs) present α-galactosylceramide (αGC) to invariant natural killer T (iNKT) cells through CD1d. Among cDC subsets, CD8(+) DCs efficiently induce IFN-γ production in iNKT cells. Using fluorescence-labeled αGC, we showed that CD8(+) DCs incorporated larger amounts of αGC and kept it intact longer than CD8(-) DCs. Histological analyses revealed that Langerin(+)CD8(+) DCs in the splenic marginal zone, which was the unique equipment to capture blood-borne antigens, preferably incorporated αGC, and the depletion of Langerin(+) cells decreased IFN-γ and IL-12 production in response to αGC. Furthermore, splenic Langerin(+)CD8(+) DCs expressed more membrane-bound CXCL16, which possibly anchored iNKT cells in the marginal zone, than CD8(-) DCs. Collectively, it is suggested that the cellular properties and localization of CD8(+) DCs are important for stimulation of iNKT cells by αGC.
Asunto(s)
Células Dendríticas/inmunología , Fluorescencia , Galactosilceramidas/inmunología , Animales , Antígenos de Superficie/inmunología , Antígenos de Superficie/metabolismo , Antígenos CD8/inmunología , Antígenos CD8/metabolismo , Carbocianinas/química , Quimiocina CXCL16 , Quimiocina CXCL6/inmunología , Quimiocina CXCL6/metabolismo , Técnicas de Cocultivo , Células Dendríticas/citología , Células Dendríticas/metabolismo , Citometría de Flujo , Colorantes Fluorescentes/química , Galactosilceramidas/metabolismo , Galactosilceramidas/farmacocinética , Interferón gamma/inmunología , Interferón gamma/metabolismo , Interleucina-12/inmunología , Interleucina-12/metabolismo , Lectinas Tipo C/inmunología , Lectinas Tipo C/metabolismo , Lectinas de Unión a Manosa/inmunología , Lectinas de Unión a Manosa/metabolismo , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Ratones Noqueados , Microscopía Fluorescente , Células T Asesinas Naturales/citología , Células T Asesinas Naturales/inmunología , Células T Asesinas Naturales/metabolismo , Bazo/citología , Bazo/inmunología , Bazo/metabolismoRESUMEN
The use of beneficial microorganisms, the so-called probiotics, to improve human health is gaining popularity. However, not all of the probiotic strains trigger the same responses and they differ in their interaction with the host. In spite of the limited knowledge on mechanisms of action some of the probiotic effects seem to be exerted through maintenance of the gastrointestinal barrier function and modulation of the immune system. In the present work, we have addressed in vitro the response of the intestinal epithelial cell line HT29 to the strain Bifidobacterium breve IPLA20004. In the array of 84 genes involved in inflammation tested, the expression of 12 was modified by the bifidobacteria. The genes of chemokine CXCL6, the chemokine receptor CCR7, and, specially, the complement component C3 were upregulated. Indeed, HT29 cells cocultivated with B. breve produced significantly higher levels of protein C3a. The proteome of HT29 cells showed increased levels of cytokeratin-8 in the presence of B. breve. Altogether, it seems that B. breve IPLA20004 could favor the recruitment of innate immune cells to the mucosa reinforcing, as well as the physical barrier of the intestinal epithelium.
Asunto(s)
Bifidobacterium/inmunología , Factores Inmunológicos/genética , Factores Inmunológicos/inmunología , Mucosa Intestinal/microbiología , Proteoma/genética , Proteoma/inmunología , Línea Celular Tumoral , Quimiocina CXCL6/genética , Quimiocina CXCL6/inmunología , Complemento C3a/genética , Complemento C3a/inmunología , Células Epiteliales/inmunología , Células Epiteliales/microbiología , Células HT29 , Humanos , Factores Inmunológicos/metabolismo , Inflamación/genética , Inflamación/inmunología , Inflamación/microbiología , Mucosa Intestinal/inmunología , Queratina-8/genética , Queratina-8/inmunología , Probióticos/metabolismo , Receptores CCR7/genética , Receptores CCR7/inmunologíaRESUMEN
Interleukin-22 (IL-22) plays a critical role in mucosal defense, although the molecular mechanisms that ensure IL-22 tissue distribution remain poorly understood. We show that the CXCL16-CXCR6 chemokine-chemokine receptor axis regulated group 3 innate lymphoid cell (ILC3) diversity and function. CXCL16 was constitutively expressed by CX3CR1(+) intestinal dendritic cells (DCs) and coexpressed with IL-23 after Citrobacter rodentium infection. Intestinal ILC3s expressed CXCR6 and its ablation generated a selective loss of the NKp46(+) ILC3 subset, a depletion of intestinal IL-22, and the inability to control C. rodentium infection. CD4(+) ILC3s were unaffected by CXCR6 deficiency and remained clustered within lymphoid follicles. In contrast, the lamina propria of Cxcr6(-/-) mice was devoid of ILC3s. The loss of ILC3-dependent IL-22 epithelial stimulation reduced antimicrobial peptide expression that explained the sensitivity of Cxcr6(-/-) mice to C. rodentium. Our results delineate a critical CXCL16-CXCR6 crosstalk that coordinates the intestinal topography of IL-22 secretion required for mucosal defense.
Asunto(s)
Quimiocina CXCL6/inmunología , Infecciones por Enterobacteriaceae/inmunología , Interleucinas/inmunología , Membrana Mucosa/inmunología , Receptores CXCR/inmunología , Animales , Antígenos Ly/biosíntesis , Linfocitos T CD4-Positivos/inmunología , Receptor 1 de Quimiocinas CX3C , Quimiocina CXCL16 , Quimiocina CXCL6/biosíntesis , Citrobacter rodentium/inmunología , Células Dendríticas/inmunología , Interleucina-23/biosíntesis , Interleucinas/biosíntesis , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Receptor 1 Gatillante de la Citotoxidad Natural/biosíntesis , Receptores CXCR/biosíntesis , Receptores CXCR/genética , Receptores CXCR6 , Receptores de Quimiocina/biosíntesis , Receptores de Quimiocina/inmunología , Interleucina-22RESUMEN
IPF is a chronic, progressive pulmonary disease, leading to respiratory failure. In search of mechanisms of IPF, we used the bleomycin-induced lung-injury model in mice, which causes acute inflammation that may progress to chronic lung inflammation and fibrosis. Here, we asked whether CXCL6/GCP-2, a member of the CXC chemokine superfamily, may be involved in IPF development. First, we reported an increase of CXCL6 levels in BALF from patients with IPF, as well as in the lung of mice, 24 h after bleomycin administration. To investigate whether CXCL6 played a role in experimental bleomycin-induced pulmonary fibrosis, we treated mice with an anti-mCXCL6 mAb that has been shown to inhibit neutrophil chemotaxis in vitro. CXCL6 antibody blockade attenuated acute inflammation with a reduced pulmonary neutrophil influx, IL-1ß, CXCL1, and TIMP-1 production. In the later phase (14 days after bleomycin exposure), lymphocyte recruitment and fibrosis markers, such as collagen and TIMP-1, were diminished, as well as collagen deposition and fibrotic lesion the lung. Therefore, the data suggest that CXCL6 contributes to experimental pulmonary fibrosis, and CXCL6 inhibition might be used to reduce lung toxicity associated with bleomycin treatment.
Asunto(s)
Antibióticos Antineoplásicos/efectos adversos , Anticuerpos Monoclonales de Origen Murino/farmacología , Anticuerpos Neutralizantes/farmacología , Bleomicina/efectos adversos , Quimiocina CXCL6/antagonistas & inhibidores , Neumonía/inmunología , Fibrosis Pulmonar/inmunología , Animales , Antibióticos Antineoplásicos/farmacología , Anticuerpos Monoclonales de Origen Murino/inmunología , Anticuerpos Neutralizantes/inmunología , Bleomicina/farmacología , Inhibición de Migración Celular/efectos de los fármacos , Inhibición de Migración Celular/inmunología , Quimiocina CXCL6/inmunología , Quimiocina CXCL6/metabolismo , Quimiotaxis de Leucocito/efectos de los fármacos , Quimiotaxis de Leucocito/inmunología , Modelos Animales de Enfermedad , Femenino , Humanos , Masculino , Ratones , Neutrófilos/inmunología , Neutrófilos/metabolismo , Neutrófilos/patología , Neumonía/inducido químicamente , Neumonía/tratamiento farmacológico , Neumonía/patología , Fibrosis Pulmonar/inducido químicamente , Fibrosis Pulmonar/tratamiento farmacológico , Fibrosis Pulmonar/patologíaRESUMEN
Relapse and graft-versus-host disease remain major problems associated with allogeneic bone marrow (BM) transplantation (allo-BMT) and posttransplantation therapy in patients with multiple myeloma (MM) and other hematologic malignancies. A possible strategy for selectively enhancing the graft-versus-myeloma response and possibly reducing graft-versus-host disease is to increase the migration of alloreactive T cells toward the MM-containing BM. In the present study, we characterized the BM-homing behavior of donor-derived effector T cells in a novel allo-BMT model for the treatment of MM. We observed that posttransplantation immunotherapy consisting of donor lymphocyte infusion (DLI) and vaccination with minor histocompatibility antigen-loaded dendritic cells (DCs) was associated with prolonged survival compared with allo-BMT with no further treatment. Moreover, CD8(+) effector T cells expressing inflammatory homing receptors, including high levels of CD44, LFA-1, and inflammatory chemokine receptors, were recruited to MM-bearing BM. This was paralleled by strongly increased expression of IFN-γ and IFN-γ-inducible chemokines, including CXCL9, CXCL10, and CXCL16, especially in mice treated with DLI plus minor histocompatibility antigen-loaded DC vaccination. Remarkably, expression of the homeostatic chemokine CXCL12 was reduced. Furthermore, IFN-γ and TNF-α induced BM endothelial cells to express high levels of the inflammatory chemokines and reduced or unaltered levels of CXCL12. Finally, presentation of CXCL9 by multiple BM endothelial cell-expressed heparan sulfate proteoglycans triggered transendothelial migration of effector T cells. Taken together, our data demonstrate that both post-transplantation DLI plus miHA-loaded DC vaccination and MM growth result in an increased expression of inflammatory homing receptors on donor T cells, decreased levels of the homeostatic BM-homing chemokine CXCL12, and strong induction of inflammatory chemokines in the BM. Thus, along with increasing the population of alloreactive T cells, post-transplantation immunotherapy also might contribute to a more effective graft-versus-tumor response by switching homeostatic T cell migration to inflammation-driven migration.
Asunto(s)
Trasplante de Médula Ósea , Médula Ósea/inmunología , Células Dendríticas/inmunología , Inmunoterapia , Mieloma Múltiple/terapia , Linfocitos T Citotóxicos/inmunología , Animales , Médula Ósea/patología , Movimiento Celular/inmunología , Quimiocina CXCL10/agonistas , Quimiocina CXCL10/biosíntesis , Quimiocina CXCL10/inmunología , Quimiocina CXCL12/antagonistas & inhibidores , Quimiocina CXCL12/biosíntesis , Quimiocina CXCL12/inmunología , Quimiocina CXCL16 , Quimiocina CXCL6/agonistas , Quimiocina CXCL6/biosíntesis , Quimiocina CXCL6/inmunología , Quimiocina CXCL9/agonistas , Quimiocina CXCL9/biosíntesis , Quimiocina CXCL9/inmunología , Células Dendríticas/química , Células Dendríticas/trasplante , Efecto Injerto vs Tumor/inmunología , Interferón gamma/agonistas , Interferón gamma/biosíntesis , Interferón gamma/inmunología , Transfusión de Linfocitos , Ratones , Antígenos de Histocompatibilidad Menor/química , Antígenos de Histocompatibilidad Menor/inmunología , Mieloma Múltiple/inmunología , Mieloma Múltiple/mortalidad , Mieloma Múltiple/patología , Análisis de Supervivencia , Trasplante Homólogo , Factor de Necrosis Tumoral alfa/agonistas , Factor de Necrosis Tumoral alfa/biosíntesis , Factor de Necrosis Tumoral alfa/inmunologíaRESUMEN
RATIONALE: The recently discovered chemokine CXC motif ligand 16 (CXCL16) is highly expressed in atherosclerotic lesions and is a potential pathogenic mediator in coronary artery disease. OBJECTIVE: The aim of this study was to test the role of CXCL16 on platelet activation and vascular adhesion, as well as the underlying mechanism and signaling pathway. METHODS AND RESULTS: Reverse-transcriptase polymerase chain reaction, Western blotting, confocal microscopy, and flow cytometry revealed that CXCL16-specific receptor, CXC motif receptor 6, is highly expressed in platelets. According to flow cytometry and confocal microscopy, stimulation of platelets with CXCL16 induced platelet degranulation, integrin α(IIb)ß(3) activation, and shape change. CXCL16 increased Akt phosphorylation (Thr(308)/Ser(473)), an effect abrogated by phosphatidylinositide 3-kinase inhibitors wortmannin (100 nmol/L) and LY294002 (25 µmol/L). The phosphatidylinositide 3-kinase inhibitors and Akt inhibitor SH-6 (20 µmol/L) further diminished CXCL16-induced platelet activation. CXCL16-mediated platelet degranulation, integrin α(IIb)ß(3) activation, and Akt phosphorylation were blunted in platelets lacking CXCL16-specific receptor CXC motif receptor 6. CXCL16-induced platelet activation was abrogated in Akt1- or Akt2-deficient platelets. CXCL16 enhanced platelet adhesion to endothelium in vitro after high arterial shear stress (2000(-s)) and to injured vascular wall in vivo after carotid ligation. CXCL16-induced stimulation of platelet adhesion again was prevented by phosphatidylinositide 3-kinase and Akt inhibitors. Apyrase and antagonists of platelet purinergic receptors P(2)Y(1) (MRS2179, 100 µmol/L) and especially P(2)Y(12) (Cangrelor, 10 µmol/L) blunted CXCL16-triggered platelet activation as well as CXCL16-induced platelet adhesion under high arterial shear stress in vitro and after carotid ligation in vivo. CONCLUSIONS: The inflammatory chemokine CXCL16 triggers platelet activation and adhesion via CXC motif receptor 6-dependent phosphatidylinositide 3-kinase/Akt signaling and paracrine activation, suggesting a decisive role for CXCL16 in linking vascular inflammation and thrombo-occlusive diseases.
Asunto(s)
Quimiocina CXCL6/inmunología , Quimiocina CXCL6/metabolismo , Activación Plaquetaria/inmunología , Adhesividad Plaquetaria/inmunología , Transducción de Señal/inmunología , Animales , Benzofuranos , Plaquetas/inmunología , Plaquetas/metabolismo , Quimiocina CXCL16 , Enfermedad de la Arteria Coronaria/inmunología , Enfermedad de la Arteria Coronaria/metabolismo , Endotelio Vascular/inmunología , Endotelio Vascular/metabolismo , Femenino , Humanos , Masculino , Ratones , Ratones Mutantes , Fosfatidilinositol 3-Quinasas/metabolismo , Proteínas Proto-Oncogénicas c-akt/genética , Proteínas Proto-Oncogénicas c-akt/metabolismo , Quinolinas , Receptores CXCR/genética , Receptores CXCR/metabolismo , Receptores CXCR6 , Trombosis/inmunología , Trombosis/metabolismo , Vasculitis/inmunología , Vasculitis/metabolismoRESUMEN
The chemokine granulocyte chemotactic protein (GCP)-2/CXCL6 promotes tumor growth as angiogenesis inducer and neutrophil chemoattractant. The neutralizing capacity and specificity of monoclonal mouse anti-murine (mu)GCP-2/CXCL6 antibodies were evidenced by granulocyte chemotaxis and signaling assays. The half-life of the non-antigenic antibody in the blood circulation was approximately 15 days. The titers remained constant upon weekly injection. Tumor growth and lymphogenic metastases of human melanoma over-expressing muGCP-2 were reduced in mice treated with anti-muGCP-2. Moreover, the drop in muGCP-2 antibody titer correlated with the melanoma tumor size. Taken together, we show that functional blocking of GCP-2 inhibits tumor growth and metastases.
Asunto(s)
Anticuerpos Neutralizantes/farmacología , Proliferación Celular/efectos de los fármacos , Quimiocina CXCL6/inmunología , Melanoma/prevención & control , Animales , Anticuerpos Monoclonales/inmunología , Anticuerpos Monoclonales/farmacocinética , Anticuerpos Monoclonales/farmacología , Anticuerpos Neutralizantes/inmunología , Línea Celular Tumoral , Quimiocina CXCL6/genética , Quimiocina CXCL6/metabolismo , Quimiotaxis de Leucocito/efectos de los fármacos , Femenino , Granulocitos/efectos de los fármacos , Granulocitos/metabolismo , Humanos , Melanoma/genética , Melanoma/patología , Melanoma Experimental/genética , Melanoma Experimental/metabolismo , Melanoma Experimental/prevención & control , Ratones , Ratones Endogámicos , Ratones Desnudos , Metástasis de la Neoplasia , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
BACKGROUND: CXCL16 mediates adhesion and phagocytosis of both Gram-negative and Gram-positive bacteria and is a strong chemoattractant for CXCR6+ T cells. In this study, we determined the so far unknown expression and signal transduction of the novel CXCL16-CXCR6 chemokine-ligand receptor system in intestinal inflammation in vivo and in vitro. METHODS: CXCL16 mRNA was measured by quantitative PCR in human colonic biopsies of patients with Crohn's disease (CD) as well as in the TNFΔARE mouse model of ileitis and in murine cytomegalovirus (MCMV)-induced colitis. CXCL16 serum levels were analyzed by ELISA. CXCL16-induced signal transduction was analyzed in intestinal epithelial cells with phospho-specific antibodies for mitogen-activated protein (MAP) kinases and Akt. RESULTS: We found an inverse expression pattern of CXCL16 and CXCR6, with highest CXCL16 mRNA expression in the proximal murine small intestine and the highest CXCR6 mRNA expression in the distal colon. CXCL16 and CXCR6 mRNA were expressed in colorectal cancer (CRC)-derived intestinal epithelial cell (IEC) lines. CRC-expressed CXCR6 was functional, as demonstrated by CXCL16-induced MAP kinase and Akt activation. Intestinal CXCL16 expression was elevated in the TNFΔARE mouse model of ileitis and in MCMV-induced colitis (P < 0.05) and in the sera and colons of patients with CD (P < 0.05), where its expression correlated highly with CXCR6 and IL-8 levels (r = 0.85 and 0.89, respectively). CONCLUSIONS: CRC-derived IECs express the functional CXCL16 receptor CXCR6. CXCL16 mRNA and protein expression is up-regulated in intestinal inflammation in vitro and in CD patients, suggesting an important role for this chemokine in intestinal inflammation.
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Quimiocina CXCL6/metabolismo , Quimiocinas CXC/metabolismo , Colitis/metabolismo , Enfermedad de Crohn/metabolismo , Receptores Depuradores/metabolismo , Animales , Línea Celular , Quimiocina CXCL16 , Quimiocina CXCL6/sangre , Quimiocina CXCL6/inmunología , Quimiocinas CXC/sangre , Quimiocinas CXC/inmunología , Colitis/inmunología , Colitis/patología , Colitis/virología , Colon/inmunología , Colon/metabolismo , Enfermedad de Crohn/inmunología , Enfermedad de Crohn/patología , Enfermedad de Crohn/virología , Infecciones por Herpesviridae/inmunología , Infecciones por Herpesviridae/metabolismo , Infecciones por Herpesviridae/patología , Humanos , Ileítis/inmunología , Ileítis/metabolismo , Ileítis/patología , Ratones , Proteínas Quinasas Activadas por Mitógenos/análisis , Proteínas Quinasas Activadas por Mitógenos/metabolismo , Muromegalovirus/inmunología , Muromegalovirus/metabolismo , Proteínas Proto-Oncogénicas c-akt/inmunología , Proteínas Proto-Oncogénicas c-akt/metabolismo , Receptores CXCR/inmunología , Receptores CXCR/metabolismo , Receptores CXCR6 , Receptores de Quimiocina/inmunología , Receptores de Quimiocina/metabolismo , Receptores Depuradores/sangre , Receptores Depuradores/inmunología , Receptores Virales/inmunología , Receptores Virales/metabolismo , Linfocitos T/inmunología , Linfocitos T/metabolismo , Regulación hacia ArribaRESUMEN
In this study, we analyzed the expression and function of CXCL16 in pneumococcal meningitis. CXCL16 was found to be up‐regulated in RAW264.7 macrophages (but not in neutrophils and endothelial cells) upon pneumococcal stimulation, in the cerebrospinal fluid of patients, and in the brains as well as the cerebrospinal fluid of mice with pneumococcal meningitis. CXCL16 up‐regulation in vivo was dependent on Toll‐like receptor (TLR) 2/TLR4 and MyD88 signaling. Neutralization of CXCL16 in animals before intracisternal pneumococcal infection (using anti‐CXCL16 antibodies) resulted in reduced cerebrospinal fluid pleocytosis. In vitro, murine neutrophils expressed the CXCL16 receptor CXCR6 and showed dose‐dependant migration toward a CXCL16 gradient. Thus, this study implicates CXCL16 as an additional neutrophil chemoattractant in cerebrospinal fluid in early pneumococcal meningitis.
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Líquido Cefalorraquídeo/citología , Quimiocina CXCL6/inmunología , Meningitis Neumocócica/inmunología , Infiltración Neutrófila , Neutrófilos/inmunología , Animales , Encéfalo/citología , Encéfalo/inmunología , Línea Celular , Líquido Cefalorraquídeo/inmunología , Quimiocina CXCL16 , Células Endoteliales/inmunología , Perfilación de la Expresión Génica , Humanos , Macrófagos/inmunología , Ratones , Ratones Endogámicos C57BL , Factor 88 de Diferenciación Mieloide/inmunología , Receptores CXCR/inmunología , Receptores CXCR6 , Receptor Toll-Like 2/inmunología , Receptor Toll-Like 4/inmunologíaRESUMEN
OBJECTIVE: To better define the activity of soluble CXCL16 in the recruitment of polymorphonuclear neutrophils (PMNs) in vivo, utilizing a novel animal model of gout involving engraftment of SCID mice with normal human synovial tissue (ST) injected intragraft with gouty human synovial fluid (SF). METHODS: For in vitro studies, a modified Boyden chemotaxis system was used to identify CXCL16 as an active recruitment factor for PMNs in gouty SF. Migration of PMNs could be reduced by neutralization of CXCL16 activity in gouty SF. For in vivo analyses, fluorescent dye-tagged PMNs were injected intravenously into SCID mice while, simultaneously, diluted gouty SF containing CXCL16, or depleted of CXCL16 by antibody blocking, was administered intragraft. In addition, the receptor for CXCL16, CXCR6, was inhibited by incubating PMNs with a neutralizing anti-CXCR6 antibody prior to injection into the mouse chimeras. Recruitment of PMNs to the gouty SF-injected normal human ST was then examined in this SCID mouse chimera system. RESULTS: CXCL16 concentrations were highly elevated in gouty SF, and PMNs were observed to migrate in response to CXCL16 in vitro. Normal human ST-SCID mouse chimeras injected intragraft with gouty SF that had been depleted of CXCL16 during PMN transfer showed a significant reduction of 50% in PMN recruitment to engrafted tissue as compared with that after administration of sham-depleted gouty SF. Similar findings were achieved when PMNs were incubated with a neutralizing anti-CXCR6 antibody before injection into chimeras. CONCLUSION: Overall, the results of this study outline the effectiveness of the human-SCID mouse chimera system as a viable animal model of gout, serving to identify the primary function of CXCL16 as a significant mediator of in vivo recruitment of PMNs to gouty SF.
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Quimiocina CXCL6/metabolismo , Modelos Animales de Enfermedad , Gota/metabolismo , Neutrófilos/metabolismo , Animales , Quimiocina CXCL16 , Quimiocina CXCL6/inmunología , Quimiotaxis de Leucocito/inmunología , Ensayo de Inmunoadsorción Enzimática , Gota/inmunología , Humanos , Ratones , Ratones SCID , Neutrófilos/inmunología , Neutrófilos/trasplante , Líquido Sinovial/inmunología , Líquido Sinovial/metabolismo , Membrana Sinovial/inmunología , Membrana Sinovial/metabolismo , Trasplante HeterólogoRESUMEN
In a nasal vaccine against influenza, the activation of natural killer T (NKT) cells by intranasal coadministration of alpha-galactosylceramide (alpha-GalCer) can potently enhance protective immune responses. The results of this study show that the NKT cell-activated nasal vaccine can induce an effective cross-protection against different strains of influenza virus, including H5 type. To analyze the mechanism of NKT cell activation by this nasal vaccine, we prepared fluorescence-labeled alpha-GalCer by which we detect a direct interaction between NKT cells and alpha-GalCer-stored dendritic cells in nasal mucosa-associated tissues. Accordingly, although very few NKT cells exist at mucosa, the nasal vaccination induced a localized increase in NKT cell population, which is partly dependent on CXCL16/CXCR6. Furthermore, we found that NKT cell activation stimulates mucosal IgA production by a mechanism that is dependent on interleukin (IL)-4 production. These results strengthen the basis of nasal vaccination via NKT cell activation, which can induce immune cross-protection.
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Adyuvantes Inmunológicos/administración & dosificación , Galactosilceramidas/administración & dosificación , Virus de la Influenza A/inmunología , Vacunas contra la Influenza/administración & dosificación , Gripe Humana/prevención & control , Células T Asesinas Naturales/efectos de los fármacos , Vacunación/métodos , Administración Intranasal , Animales , Especificidad de Anticuerpos , Quimiocina CXCL16 , Quimiocina CXCL6/inmunología , Reacciones Cruzadas , Células Dendríticas/efectos de los fármacos , Células Dendríticas/inmunología , Colorantes Fluorescentes , Humanos , Inmunoglobulina A/biosíntesis , Inmunoglobulina A/inmunología , Subtipo H5N1 del Virus de la Influenza A/inmunología , Virus de la Influenza B/inmunología , Vacunas contra la Influenza/inmunología , Gripe Humana/inmunología , Gripe Humana/virología , Interleucina-4/inmunología , Activación de Linfocitos/efectos de los fármacos , Activación de Linfocitos/inmunología , Ratones , Ratones Endogámicos BALB C , Mucosa Nasal/inmunología , Células T Asesinas Naturales/inmunología , Receptores CXCR/inmunología , Receptores CXCR6RESUMEN
PROBLEM: CXCL6 is a potent pro-inflammatory neutrophil chemoattractant and activator whose activity during pregnancy is not well-established. The purpose of this study was to determine if CXCL6 is present in amniotic fluid (AF) and if CXCL6 concentrations in AF change with labor (pre-term and term) or intra-amniotic infection/inflammation (IAI). METHOD OF STUDY: A cross-sectional study was designed including the following groups: (1) mid-trimester (n = 65); (2) term no labor (n = 20); (3) term labor (n = 44); (4) patients with pre-term labor (PTL) with subsequent term delivery (n = 57); (5) PTL without IAI who delivered pre-term (n = 47); and (6) PTL with IAI (n = 62). AF CXCL6 concentrations were determined by ELISA. RESULTS: CXCL6 was present in all term samples, but undetectable in 64/65 mid-trimester specimens. Patients with PTL and IAI had a significantly higher median AF CXCL6 concentration than those with PTL without IAI [228.9 pg/mL (0.0-8344.8) versus 55.7 pg/mL (0-454.4); P < 0.05] and those with PTL and term delivery [41.5 pg/mL (0-279.0); P < 0.05]. The median AF CXCL6 concentration did not change with spontaneous term labor [term no labor: 81.1 pg/mL (8.5-201.7) versus term labor: 75.2 pg/mL (6.7-378.7): P = 0.7]. CONCLUSION: (1) CXCL6 is detectable in AF and its concentration increases with gestational age; (2) IAI results in increased AF CXCL6 concentrations, suggesting that CXCL6 plays a role in the deployment of an inflammatory response; (3) In contrast to related chemokines, specifically IL-8, AF CXCL6 does not appear to be involved in spontaneous term parturition. These observations are novel, and suggest a role for CXCL6 in the innate immune response to microbial invasion of the amniotic cavity.
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Amnios/inmunología , Líquido Amniótico/química , Quimiocina CXCL6/análisis , Inmunidad Innata , Trabajo de Parto Prematuro/inmunología , Nacimiento a Término/inmunología , Adolescente , Adulto , Amnios/metabolismo , Quimiocina CXCL6/inmunología , Estudios Transversales , Femenino , Edad Gestacional , Humanos , Trabajo de Parto Prematuro/metabolismo , Embarazo , Nacimiento a Término/metabolismoRESUMEN
Cerebral malaria (CM) results from the binding of infected erythrocytes and leukocytes to brain endothelia. The precise mechanisms underlying lymphocyte recruitment and activation in CM remain unclear. Therefore, the expression of various chemokines was quantified in brains of mice infected with Plasmodium berghei ANKA (PbA). Several chemokines attracting monocytes and activated T-lymphocytes were expressed at high levels. Their expression was almost completely abrogated in IFN-gamma ligand and receptor KO mice, indicating that IFN-gamma is an essential chemokine inducer in vivo. Surprisingly, the expression levels of chemokines, IFN-gamma and also adhesion molecules in the brain were not lower in CM-resistant Balb/c and DBA/2 mice compared to CM-sensitive C57BL/6 and DBA/1 mice, although T lymphocyte sequestration in the brain was significantly less in CM-resistant than in CM-sensitive mice. This difference correlated with a higher up-regulation of the CXC chemokine receptor (CXCR)-3 on splenic T cells and a higher chemotactic response to IFN-gamma-inducible protein-10 (IP-10) in C57BL/6 compared to Balb/c mice. In conclusion, parasite-induced IFN-gamma in the brain results in high local expression levels of specific chemokines for monocytes and lymphocytes. The strain-dependent susceptibility to develop CM is more related to the expression of CXCR3 in circulating leukocytes than to the chemokine expression levels in the brain.
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Movimiento Celular/inmunología , Quimiocinas/inmunología , Interferón gamma/inmunología , Malaria Cerebral/inmunología , Receptores CXCR3/inmunología , Linfocitos T/citología , Linfocitos T/inmunología , Animales , Moléculas de Adhesión Celular/metabolismo , Células Cultivadas , Quimiocina CXCL12/genética , Quimiocina CXCL12/inmunología , Quimiocina CXCL6/genética , Quimiocina CXCL6/inmunología , Quimiocinas/genética , Quimiocinas/metabolismo , Regulación de la Expresión Génica/genética , Regulación de la Expresión Génica/inmunología , Ligandos , Macrófagos/citología , Macrófagos/inmunología , Malaria Cerebral/parasitología , Ratones , Monocitos/citología , Monocitos/inmunología , Plasmodium berghei/inmunología , Receptores de Interferón/metabolismo , Bazo/citología , Bazo/inmunología , Tasa de Supervivencia , Receptor de Interferón gammaRESUMEN
Chemokines, scavenger receptors and adhesion molecules have long been known as important players in the pathogenesis of atherosclerosis. A series of studies conducted in the past few years described CXCL16/SR-PSOX--a new molecule combining those three functions, and suggested that CXCL16/SR-PSOX can be a potential player in atherogenesis. Initial ex vivo studies showed that CXCL16/SR-PSOX is abundant in human and murine atherosclerotic lesions. Following in vitro studies suggested that as an adhesion molecule CXCL16/SR-PSOX might mediate T-cell adhesion to the endothelium, as a chemokine - drive T-cell migration, stimulate cell proliferation and elicit inflammatory phenotype in smooth muscle cells (SMC) and, finally, as a scavenger receptor - mediate uptake of atherogenic lipoproteins by macrophages and SMC. All these effects are known to be pro-atherogenic. Surprisingly, in vivo studies performed in murine models of atherosclerosis suggested that CXCL16/SR-PSOX is atheroprotective, while its receptor CXCR6 is harmful. In addition, studies investigating the association of circulating CXCL16/SR-PSOX plasma concentrations with the presence and extent of coronary artery disease (CAD) in humans are controversial suggesting both positive, negative and no association. To finally answer the question whether CXCL16/SR-PSOX can serve as a causative factor, biomarker or even a therapeutic target in atherosclerosis, we are currently in need of carefully designed animal and human studies investigating the effects of CXCL16/SR-PSOX and CXCR6 deficiency, inhibition and over-expression on the progression of atherosclerosis. Such complex approach will help us unravel the mystery of CXCL16/SR-PSOX in atherosclerosis and hopefully develop better ways of treating atherosclerosis by targeting this interesting molecule.