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1.
J Immunol ; 198(10): 4115-4128, 2017 05 15.
Artigo em Inglês | MEDLINE | ID: mdl-28396316

RESUMO

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.


Assuntos
Arteríolas/fisiologia , Decídua/fisiologia , Células Endoteliais/metabolismo , Células Matadoras Naturais/fisiologia , Macrófagos/fisiologia , Trofoblastos/metabolismo , Arteríolas/citologia , Arteríolas/imunologia , Movimento Celular/imunologia , Células Cultivadas , Quimiocina CXCL6/biossíntese , Quimiocina CXCL6/imunologia , Quimiocinas CC/biossíntese , Quimiocinas CC/imunologia , Meios de Cultura/química , Decídua/imunologia , Células Endoteliais/efeitos dos fármacos , Células Endoteliais/imunologia , Feminino , Humanos , Interleucina-6/imunologia , Interleucina-6/metabolismo , Interleucina-8/imunologia , Interleucina-8/metabolismo , Macrófagos/imunologia , Placenta/citologia , Placenta/imunologia , Gravidez , Primeiro Trimestre da Gravidez , Trofoblastos/imunologia
2.
Gene ; 593(1): 76-83, 2016 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-27520585

RESUMO

The transcription factor PITX2 is implicated in glaucoma pathology. In an earlier study we had used microarray analysis to identify genes in the trabecular meshwork (TM) that are affected by knock down of PITX2. Here, those studies were pursued to identify genes that are direct targets of PITX2 and that may be relevant to glaucoma. Initially, bioinformatics tools were used to select among the genes that had been affected by PITX2 knock down those that have PITX2 binding sites and that may be involved in glaucoma related functions. Subsequently, the effect of PITX2 was tested using the dual luciferase assay in four cell cultures including two primary TM cultures co-transfected with vectors containing promoter fragments of six candidate genes upstream of a luciferase gene and a vector that expressed PITX2. Finally, the effect of PITX2 on endogenous expression of two genes was assessed by over expression and knock down of PITX2 in TM cells. Thirty four genes were found to contain PITX2 binding sites in their putative promoter regions, and 16 were found to be associated with TM-specific and/or glaucoma associated functions. Results of dual luciferase assays confirmed that two of six genes tested were directly targeted by PITX2. The two genes were CXCL6 (chemokine (C-X-C motif) ligand 6) and BBS5 (Bardet-Biedl syndrome 5). Over expression and knock down of PITX2 showed that this transcription factor affects endogenous expression of these two genes in TM cells. CXCL6 encodes a pro-inflammatory cytokine, and many studies have suggested that cytokines and other immune system functions are involved in glaucoma pathogenesis. BBS5 is a member of the BBS family of genes that affect ciliary functions, and ciliary bodies in the anterior chamber of the eye produce the aqueous fluid that affects intraocular pressure. Immune related functions and intraocular pressure are both important components of glaucoma pathology. The role of PITX2 in glaucoma may be mediated partly by regulating the expression of CXCL6 and BBS5 and thus affecting immune functions and intraocular pressure.


Assuntos
Câmara Anterior/metabolismo , Quimiocina CXCL6/biossíntese , Proteínas do Olho/metabolismo , Regulação da Expressão Gênica , Glaucoma/metabolismo , Proteínas de Homeodomínio/metabolismo , Proteínas/metabolismo , Elementos de Resposta , Fatores de Transcrição/metabolismo , Quimiocina CXCL6/genética , Cílios/genética , Cílios/metabolismo , Proteínas do Citoesqueleto , Proteínas do Olho/genética , Feminino , Glaucoma/genética , Glaucoma/patologia , Proteínas de Homeodomínio/genética , Humanos , Pressão Intraocular/genética , Masculino , Proteínas de Ligação a Fosfato , Proteínas/genética , Fatores de Transcrição/genética , Proteína Homeobox PITX2
3.
Immunity ; 41(5): 776-88, 2014 Nov 20.
Artigo em Inglês | MEDLINE | ID: mdl-25456160

RESUMO

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.


Assuntos
Quimiocina CXCL6/imunologia , Infecções por Enterobacteriaceae/imunologia , Interleucinas/imunologia , Mucosa/imunologia , Receptores CXCR/imunologia , Animais , Antígenos Ly/biossíntese , Linfócitos T CD4-Positivos/imunologia , Receptor 1 de Quimiocina CX3C , Quimiocina CXCL16 , Quimiocina CXCL6/biossíntese , Citrobacter rodentium/imunologia , Células Dendríticas/imunologia , Interleucina-23/biossíntese , Interleucinas/biossíntese , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Receptor 1 Desencadeador da Citotoxicidade Natural/biossíntese , Receptores CXCR/biossíntese , Receptores CXCR/genética , Receptores CXCR6 , Receptores de Quimiocinas/biossíntese , Receptores de Quimiocinas/imunologia , Interleucina 22
4.
PLoS One ; 9(11): e112327, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25372401

RESUMO

Non-alcoholic fatty liver disease (NAFLD) is a major cause of morbidity and mortality in developed countries, resulting in steatohepatitis (NASH), fibrosis and eventually cirrhosis. Modulating inflammatory mediators such as chemokines may represent a novel therapeutic strategy for NAFLD. We recently demonstrated that the chemokine receptor CXCR6 promotes hepatic NKT cell accumulation, thereby controlling inflammation in experimental NAFLD. In this study, we first investigated human biopsies (n = 20), confirming that accumulation of inflammatory cells such as macrophages is a hallmark of progressive NAFLD. Moreover, CXCR6 gene expression correlated with the inflammatory activity (ALT levels) in human NAFLD. We then tested the hypothesis that pharmacological inhibition of CXCL16 might hold therapeutic potential in NAFLD, using mouse models of acute carbon tetrachloride (CCl4)- and chronic methionine-choline-deficient (MCD) diet-induced hepatic injury. Neutralizing CXCL16 by i.p. injection of anti-CXCL16 antibody inhibited the early intrahepatic NKT cell accumulation upon acute toxic injury in vivo. Weekly therapeutic anti-CXCL16 administrations during the last 3 weeks of 6 weeks MCD diet significantly decreased the infiltration of inflammatory macrophages into the liver and intrahepatic levels of inflammatory cytokines like TNF or MCP-1. Importantly, anti-CXCL16 treatment significantly reduced fatty liver degeneration upon MCD diet, as assessed by hepatic triglyceride levels, histological steatosis scoring and quantification of lipid droplets. Moreover, injured hepatocytes up-regulated CXCL16 expression, indicating that scavenging functions of CXCL16 might be additionally involved in the pathogenesis of NAFLD. Targeting CXCL16 might therefore represent a promising novel therapeutic approach for liver inflammation and steatohepatitis.


Assuntos
Quimiocina CXCL6/biossíntese , Quimiocinas CXC/biossíntese , Fígado/metabolismo , Macrófagos/metabolismo , Hepatopatia Gordurosa não Alcoólica/metabolismo , Receptores Depuradores/biossíntese , Regulação para Cima , Animais , Intoxicação por Tetracloreto de Carbono/metabolismo , Intoxicação por Tetracloreto de Carbono/patologia , Quimiocina CXCL16 , Doença Crônica , Feminino , Hepatócitos/metabolismo , Hepatócitos/patologia , Humanos , Fígado/patologia , Macrófagos/patologia , Masculino , Camundongos , Hepatopatia Gordurosa não Alcoólica/patologia
5.
J Immunol ; 191(8): 4202-10, 2013 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-24038090

RESUMO

During infection with the helminth parasite Schistosoma mansoni, Ab regulates hepatic inflammation, and local production of Ig in the liver appears to play a role in this process. Exploring the development of the B cell response during infection, we found that parasite-specific IgG1-secreting plasma cells appeared first in the hepatic and mesenteric lymph nodes (LNs) and then at later times in the spleen, liver, and bone marrow. The LN B cell population peaked between weeks 10 and 12 of infection, and then contracted at a time that coincided with the expansion of the hepatic IgG1(+) B cell compartment, suggesting that B cells migrate from LNs to liver. CXCL9 and -16 expression in the liver increased during the time frame of B cell recruitment. Expression of the CXCL16 receptor CXCR6 was increased on B cells within the hepatic LNs, but not the mesenteric LNs. CXCR3, the receptor for CXCL9, was broadly expressed on IgG1(+) B cells in LNs and liver during infection. Increased hepatic expression of CXCL9 and -16 failed to occur if the IL-10R was blocked in vivo, an intervention associated with decreased liver B cell infiltration and the development of severe disease. Hepatic LN IgG1(+) cells migrated toward CXCL9 and -16 in vitro and to the liver in a pertussis toxin-sensitive fashion. Our data suggest that the coordinated expression of CXCL9 and -16 in the liver and of CXCR6 and CXCR3 on responding B cells within the hepatic LNs underpins establishment of the hepatic B cell infiltrate during chronic schistosomiasis.


Assuntos
Linfócitos B/imunologia , Imunoglobulina G/imunologia , Fígado/imunologia , Schistosoma mansoni/imunologia , Esquistossomose mansoni/imunologia , Transferência Adotiva , Animais , Medula Óssea/imunologia , Movimento Celular/imunologia , Quimiocina CXCL16 , Quimiocina CXCL6/biossíntese , Quimiocina CXCL9/biossíntese , Inflamação/imunologia , Fígado/citologia , Linfonodos/citologia , Linfonodos/imunologia , Ativação Linfocitária/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Transgênicos , Toxina Pertussis , Receptores CXCR/biossíntese , Receptores CXCR3/biossíntese , Receptores CXCR6 , Receptores de Interleucina-10/biossíntese , Esquistossomose mansoni/parasitologia , Baço/citologia , Baço/imunologia
6.
J Immunol ; 190(10): 5226-36, 2013 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-23596313

RESUMO

Chronic liver injury characteristically results in hepatic inflammation, which represents a prerequisite for organ fibrosis. Although NKT cells are abundantly present in liver and involved in hepatic inflammation, molecular mechanisms of their recruitment in liver fibrosis remained elusive. We hypothesized that chemokine receptor CXCR6 and its ligand CXCL16 control NKT cell migration and functionality in liver fibrosis. In patients with chronic liver diseases (n = 58), CXCR6 and CXCL16 expression was intrahepatically upregulated compared with controls. In murine liver, Cxcl16 was strongly expressed by endothelium and macrophages, whereas lymphocyte populations (NKT, NK, CD4 T, CD8 T cells) expressed CXCR6. Intravital two-photon microscopy imaging of Cxcr6(+/gfp) and Cxcr6(gfp/gfp) mice and chemotaxis studies in vitro revealed that CXCR6 specifically controls hepatic NKT cell accumulation during the early response upon experimental liver damage. Hepatic invariant NKT cells expressed distinct proinflammatory cytokines including IFN-γ and IL-4 upon injury. CXCR6-deficient mice were protected from liver fibrosis progression in two independent experimental models. Macrophage infiltration and protein levels of inflammatory cytokines IFN-γ, TNF-α, and IL-4 were also reduced in fibrotic livers of Cxcr6(-/-) mice, corroborating that hepatic NKT cells provide essential cytokine signals perpetuating hepatic inflammation and fibrogenesis. Adoptive transfer of NKT cells, but not CD4 T cells, isolated from wild type livers restored hepatic fibrosis in Cxcr6(-/-) mice upon experimental steatohepatitis. Our results demonstrate that hepatic NKT cells accumulate CXCR6-dependent early upon injury, thereby accentuating the inflammatory response in the liver and promoting hepatic fibrogenesis. Interfering with CXCR6/CXCL16 might therefore bear therapeutic potential in liver fibrosis.


Assuntos
Quimiocina CXCL6/metabolismo , Cirrose Hepática/imunologia , Células T Matadoras Naturais/imunologia , Receptores CXCR/metabolismo , Transferência Adotiva , Animais , Linfócitos T CD4-Positivos/imunologia , Movimento Celular , Células Cultivadas , Quimiocina CXCL16 , Quimiocina CXCL6/biossíntese , Quimiocina CXCL6/sangue , Fígado Gorduroso , Hepatócitos/imunologia , Humanos , Inflamação/imunologia , Interferon gama/biossíntese , Interleucina-4/biossíntese , Fígado/imunologia , Fígado/lesões , Fígado/metabolismo , Cirrose Hepática/metabolismo , Hepatopatias/metabolismo , Macrófagos/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Células T Matadoras Naturais/metabolismo , Receptores CXCR/biossíntese , Receptores CXCR/genética , Receptores CXCR6 , Regulação para Cima
7.
Biol Blood Marrow Transplant ; 19(3): 378-86, 2013 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-23266741

RESUMO

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.


Assuntos
Transplante de Medula Óssea , Medula Óssea/imunologia , Células Dendríticas/imunologia , Imunoterapia , Mieloma Múltiplo/terapia , Linfócitos T Citotóxicos/imunologia , Animais , Medula Óssea/patologia , Movimento Celular/imunologia , Quimiocina CXCL10/agonistas , Quimiocina CXCL10/biossíntese , Quimiocina CXCL10/imunologia , Quimiocina CXCL12/antagonistas & inibidores , Quimiocina CXCL12/biossíntese , Quimiocina CXCL12/imunologia , Quimiocina CXCL16 , Quimiocina CXCL6/agonistas , Quimiocina CXCL6/biossíntese , Quimiocina CXCL6/imunologia , Quimiocina CXCL9/agonistas , Quimiocina CXCL9/biossíntese , Quimiocina CXCL9/imunologia , Células Dendríticas/química , Células Dendríticas/transplante , Efeito Enxerto vs Tumor/imunologia , Interferon gama/agonistas , Interferon gama/biossíntese , Interferon gama/imunologia , Transfusão de Linfócitos , Camundongos , Antígenos de Histocompatibilidade Menor/química , Antígenos de Histocompatibilidade Menor/imunologia , Mieloma Múltiplo/imunologia , Mieloma Múltiplo/mortalidade , Mieloma Múltiplo/patologia , Análise de Sobrevida , Transplante Homólogo , Fator de Necrose Tumoral alfa/agonistas , Fator de Necrose Tumoral alfa/biossíntese , Fator de Necrose Tumoral alfa/imunologia
8.
Genes Immun ; 14(2): 67-82, 2013 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-23190644

RESUMO

The aim of this study is to understand intracellular regulatory mechanisms in peripheral blood mononuclear cells (PBMCs), which are either common to many autoimmune diseases or specific to some of them. We incorporated large-scale data such as protein-protein interactions, gene expression and demographical information of hundreds of patients and healthy subjects, related to six autoimmune diseases with available large-scale gene expression measurements: multiple sclerosis (MS), systemic lupus erythematosus (SLE), juvenile rheumatoid arthritis (JRA), Crohn's disease (CD), ulcerative colitis (UC) and type 1 diabetes (T1D). These data were analyzed concurrently by statistical and systems biology approaches tailored for this purpose. We found that chemokines such as CXCL1-3, 5, 6 and the interleukin (IL) IL8 tend to be differentially expressed in PBMCs of patients with the analyzed autoimmune diseases. In addition, the anti-apoptotic gene BCL3, interferon-γ (IFNG), and the vitamin D receptor (VDR) gene physically interact with significantly many genes that tend to be differentially expressed in PBMCs of patients with the analyzed autoimmune diseases. In general, similar cellular processes tend to be differentially expressed in PBMC in the analyzed autoimmune diseases. Specifically, the cellular processes related to cell proliferation (for example, epidermal growth factor, platelet-derived growth factor, nuclear factor-κB, Wnt/ß-catenin signaling, stress-activated protein kinase c-Jun NH2-terminal kinase), inflammatory response (for example, interleukins IL2 and IL6, the cytokine granulocyte-macrophage colony-stimulating factor and the B-cell receptor), general signaling cascades (for example, mitogen-activated protein kinase, extracellular signal-regulated kinase, p38 and TRK) and apoptosis are activated in most of the analyzed autoimmune diseases. However, our results suggest that in each of the analyzed diseases, apoptosis and chemotaxis are activated via different subsignaling pathways. Analyses of the expression levels of dozens of genes and the protein-protein interactions among them demonstrated that CD and UC have relatively similar gene expression signatures, whereas the gene expression signatures of T1D and JRA relatively differ from the signatures of the other autoimmune diseases. These diseases are the only ones activated via the Fcɛ pathway. The relevant genes and pathways reported in this study are discussed at length, and may be helpful in the diagnoses and understanding of autoimmunity and/or specific autoimmune diseases.


Assuntos
Doenças Autoimunes/genética , Doenças Autoimunes/imunologia , Leucócitos Mononucleares/imunologia , Mapas de Interação de Proteínas , Receptores de IgE/imunologia , Transcriptoma , Apoptose , Artrite Juvenil/imunologia , Artrite Juvenil/metabolismo , Doenças Autoimunes/metabolismo , Proteína 3 do Linfoma de Células B , Proliferação de Células , Quimiocina CXCL1/biossíntese , Quimiocina CXCL5/biossíntese , Quimiocina CXCL6/biossíntese , Quimiocinas CXC/biossíntese , Colite Ulcerativa/genética , Colite Ulcerativa/imunologia , Colite Ulcerativa/metabolismo , Doença de Crohn/genética , Doença de Crohn/imunologia , Doença de Crohn/metabolismo , Diabetes Mellitus Tipo 1/genética , Diabetes Mellitus Tipo 1/imunologia , Diabetes Mellitus Tipo 1/metabolismo , Expressão Gênica , Humanos , Inflamação , Interferon gama/metabolismo , Interleucina-8/biossíntese , Leucócitos Mononucleares/metabolismo , Lúpus Eritematoso Sistêmico/genética , Lúpus Eritematoso Sistêmico/imunologia , Lúpus Eritematoso Sistêmico/metabolismo , Sistema de Sinalização das MAP Quinases , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Esclerose Múltipla/genética , Esclerose Múltipla/imunologia , Esclerose Múltipla/metabolismo , Proteínas Proto-Oncogênicas/metabolismo , Receptores de Calcitriol/metabolismo , Transdução de Sinais , Fatores de Transcrição/metabolismo
9.
Oncol Rep ; 29(3): 975-82, 2013 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-23242131

RESUMO

Colorectal cancer (CRC) is a typical lifestyle-related disease, and it metastasizes mostly to the liver. It is important to understand the molecular mechanisms of CRC metastasis in order to design new and effective treatments for CRC patients. Chemokines are known to have antitumor effects as their chemoattractant properties stimulate the accumulation of infiltrating immune cells (TILs) in tumors. Chemokine (C-X-C motif) ligand 16 (CXCL16), also known as SR-PSOX, is a unique membrane-bound chemokine that induces the expression of its specific receptor CXCR6. We previously reported that the expression of CXCL16 by cancer cells enhances the recruitment of TILs, thereby improving the prognosis of CRC. It has since been reported that CXCL16/CXCR6 expression is involved in the metastasis of various types of cancer. However, there is no report of the association between CXCL16 expression and liver metastasis in CRC. In this study, we investigated the role of cancer-derived CXCL16 and the possibility of gene therapy using CXCL16. Therefore, we examined the metastasis of colon 38 SL4 cells to the liver in an experimental model. Following injection of cancer cells into the intraportal vein, CXCL16-expressing CRC cells drastically inhibited liver metastasis. We also found that CD8 T cells and natural killer T (NKT) cells, known as CXCR6-expressing cells, increased in CXCL16-expressing metastatic tissue. Collectively, the inhibitory effect on metastasis to the liver by CXCL16 was observed in NKT cell-depleted mice but not in CD8 T cell-depleted mice. These results demonstrate the inhibitory effect of CXCL16 on liver metastasis via NKT cells in CRC.


Assuntos
Quimiocina CXCL6/biossíntese , Neoplasias Colorretais/terapia , Neoplasias Hepáticas/prevenção & controle , Células T Matadoras Naturais/imunologia , Animais , Linhagem Celular Tumoral , Quimiocina CXCL16 , Quimiocina CXCL6/genética , Quimiocina CXCL6/fisiologia , Neoplasias Colorretais/imunologia , Neoplasias Colorretais/patologia , Feminino , Terapia Genética , Humanos , Neoplasias Hepáticas/imunologia , Neoplasias Hepáticas/secundário , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Nus , Transplante de Neoplasias
11.
J Immunol ; 188(5): 2093-101, 2012 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-22287719

RESUMO

CFA is a strong adjuvant capable of stimulating cellular immune responses. Paradoxically, adjuvant immunotherapy by prior exposure to CFA or live mycobacteria suppresses the severity of experimental autoimmune encephalomyelitis (EAE) and spontaneous diabetes in rodents. In this study, we investigated immune responses during adjuvant immunotherapy of EAE. Induction of EAE in CFA-pretreated mice resulted in a rapid influx into the draining lymph nodes (dLNs) of large numbers of CD11b(+)Gr-1(+) myeloid cells, consisting of immature cells with ring-shaped nuclei, macrophages, and neutrophils. Concurrently, a population of mycobacteria-specific IFN-γ-producing T cells appeared in the dLNs. Immature myeloid cells in dLNs expressed the chemokines CXCL10 and CXCL16 in an IFN-γ-dependent manner. Subsequently, CD4(+) T cells coexpressing the cognate chemokine receptors CXCR3 and CXCR6 and myelin oligodendrocyte glycoprotein (MOG)-specific CD4(+) T cells accumulated within the chemokine-expressing dLNs, rather than within the CNS. Migration of CD4(+) T cells toward dLN cells was abolished by depleting the CD11b(+) cells and was also mediated by the CD11b(+) cells alone. In addition to altering the distribution of MOG-specific T cells, adjuvant treatment suppressed development of MOG-specific IL-17. Thus, adjuvant immunotherapy of EAE requires IFN-γ, which suppresses development of the Th17 response, and diverts autoreactive T cells away from the CNS toward immature myeloid cells expressing CXCL10 and CXCL16 in the lymph nodes.


Assuntos
Diferenciação Celular/imunologia , Quimiocina CXCL10/biossíntese , Quimiocina CXCL6/biossíntese , Encefalomielite Autoimune Experimental/imunologia , Encefalomielite Autoimune Experimental/terapia , Proteínas da Mielina/imunologia , Células Mieloides/imunologia , Receptores CXCR3/biossíntese , Receptores CXCR/biossíntese , Subpopulações de Linfócitos T/imunologia , Animais , Inibição de Migração Celular/imunologia , Quimiocina CXCL16 , Encefalomielite Autoimune Experimental/microbiologia , Feminino , Adjuvante de Freund/administração & dosagem , Adjuvante de Freund/uso terapêutico , Interferon gama/fisiologia , Interferon gama/uso terapêutico , Linfonodos/imunologia , Linfonodos/microbiologia , Linfonodos/patologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Mycobacterium tuberculosis/imunologia , Glicoproteína Mielina-Oligodendrócito , Células Mieloides/microbiologia , Células Mieloides/patologia , Receptores CXCR6 , Subpopulações de Linfócitos T/metabolismo , Subpopulações de Linfócitos T/patologia
12.
Mol Vis ; 17: 53-62, 2011 Jan 08.
Artigo em Inglês | MEDLINE | ID: mdl-21245951

RESUMO

PURPOSE: The biologic relevance of human connective tissue growth factor (hCTGF) for primary human tenon fibroblasts (HTFs) was investigated by RNA expression profiling using affymetrix(TM) oligonucleotide array technology to identify genes that are regulated by hCTGF. METHODS: Recombinant hCTGF was expressed in HEK293T cells and purified by affinity and gel chromatography. Specificity and biologic activity of hCTGF was confirmed by biosensor interaction analysis and proliferation assays. For RNA expression profiling HTFs were stimulated with hCTGF for 48h and analyzed using affymetrix(TM) oligonucleotide array technology. Results were validated by real time RT-PCR. RESULTS: hCTGF induces various groups of genes responsible for a wound healing and inflammatory response in HTFs. A new subset of CTGF inducible inflammatory genes was discovered (e.g., chemokine [C-X-C motif] ligand 1 [CXCL1], chemokine [C-X-C motif] ligand 6 [CXCL6], interleukin 6 [IL6], and interleukin 8 [IL8]). We also identified genes that can transmit the known biologic functions initiated by CTGF such as proliferation and extracellular matrix remodelling. Of special interest is a group of genes, e.g., osteoglycin (OGN) and osteomodulin (OMD), which are known to play a key role in osteoblast biology. CONCLUSIONS: This study specifies the important role of hCTGF for primary tenon fibroblast function. The RNA expression profile yields new insights into the relevance of hCTGF in influencing biologic processes like wound healing, inflammation, proliferation, and extracellular matrix remodelling in vitro via transcriptional regulation of specific genes. The results suggest that CTGF potentially acts as a modulating factor in inflammatory and wound healing response in fibroblasts of the human eye.


Assuntos
Fator de Crescimento do Tecido Conjuntivo/metabolismo , Fibroblastos/metabolismo , Perfilação da Expressão Gênica , Regulação da Expressão Gênica , Cicatrização , Quimiocina CXCL1/biossíntese , Quimiocina CXCL6/biossíntese , Proteínas da Matriz Extracelular/biossíntese , Humanos , Técnicas In Vitro , Inflamação , Peptídeos e Proteínas de Sinalização Intercelular/biossíntese , Interleucina-6/biossíntese , Interleucina-8/biossíntese , Proteoglicanas/biossíntese , Proteínas Recombinantes/química , Reação em Cadeia da Polimerase Via Transcriptase Reversa
13.
Cytokine ; 53(3): 320-6, 2011 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-21177121

RESUMO

BACKGROUND: CXCL16 has been shown to be involved in atherosclerotic lesion development, but its role in preexisting lesions is still unclear. This study aims to assess the effect of CXCL16 on the stability of preexisting lesions. METHODS: We firstly measured plasma CXCL16 level in Apolipoprotein E-Knockout (ApoE KO) mice with either high-cholesterol diet (HCD) or normal diet (ND) by enzyme-linked immunosorbent assay (ELISA). Then, silastic collars were placed around the carotid arteries in HCD-ApoE KO mice to accelerate atherosclerotic lesions. Five weeks later, CXCL16 was overexpressed by intravenous injection of lentivirus carrying CXCL16 transgene. Two weeks after infection, lesions were stained with hematoxylin and eosin (HE) and with oil red O. Biomarkers in the lesions, such as MMPs, CCL2, VCAM-1 and TNF-α were measured by real-time polymerase chain reaction (RT-PCR), which indicate the instability of plaques. RESULTS: The level of CXCL16 in plasma was higher in HCD-ApoE KO mice as compared to ND-ApoE KO mice. Circulating CXCL16 overexpression does not affect the size of preexisting plaques, but it leads to vulnerable plaque morphology and increases the expression of markers of plaque destabilization. CONCLUSION: Systemic CXCL16 becomes much higher in atherosclerosis, and it could be a potential atherogenic biomarker. Overexpression of CXCL16 promotes the evolution of preexisting lesions to vulnerable plaques in ApoE KO mice.


Assuntos
Apolipoproteínas E/deficiência , Artérias Carótidas/metabolismo , Quimiocina CXCL6/sangue , Placa Aterosclerótica/sangue , Animais , Apolipoproteínas E/genética , Braquetes/efeitos adversos , Artérias Carótidas/patologia , Quimiocina CCL2/genética , Quimiocina CXCL16 , Quimiocina CXCL6/biossíntese , Quimiocina CXCL6/genética , Colesterol na Dieta/administração & dosagem , Colesterol na Dieta/efeitos adversos , Ensaio de Imunoadsorção Enzimática , Expressão Gênica , Masculino , Metaloproteinases da Matriz/genética , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Fenótipo , Placa Aterosclerótica/etiologia , Placa Aterosclerótica/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transfecção , Fator de Necrose Tumoral alfa/genética , Molécula 1 de Adesão de Célula Vascular/genética
14.
J Immunol Methods ; 352(1-2): 89-100, 2010 Jan 31.
Artigo em Inglês | MEDLINE | ID: mdl-19800886

RESUMO

The mouse spinal cord is an important site for autoimmune and injury models. Skull thinning surgery provides a minimally invasive window for microscopy of the mouse cerebral cortex, but there are no parallel methods for the spinal cord. We introduce a novel, facile and inexpensive method for two-photon laser scanning microscopy of the intact spinal cord in the mouse by taking advantage of the naturally accessible intervertebral space. These are powerful methods when combined with gene-targeted mice in which endogenous immune cells are labeled with green fluorescent protein (GFP). We first demonstrate that generation of the intervertebral window does not elicit a reaction of GFP(+) microglial cells in CX3CR1(gfp/+) mice. We next demonstrate a distinct rostrocaudal migration of GFP(+) immune cells in the spinal cord of CXCR6(gfp/+) mice during active experimental autoimmune encephalomyelitis (EAE). Interestingly, infiltration of the cerebral cortex by GFP(+) cells in these mice required three conditions: EAE induction, cortical injury and expression of CXCR6 on immune cells.


Assuntos
Córtex Cerebral/imunologia , Encefalomielite Autoimune Experimental/imunologia , Receptores CXCR/biossíntese , Medula Espinal/imunologia , Linfócitos T/metabolismo , Animais , Movimento Celular , Córtex Cerebral/lesões , Córtex Cerebral/patologia , Quimiocina CXCL16 , Quimiocina CXCL6/biossíntese , Quimiocina CXCL6/genética , Encefalomielite Autoimune Experimental/diagnóstico , Encefalomielite Autoimune Experimental/patologia , Proteínas de Fluorescência Verde/genética , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Microscopia Confocal , Procedimentos Cirúrgicos Minimamente Invasivos , Fótons , Receptores CXCR/genética , Receptores CXCR6 , Medula Espinal/patologia , Linfócitos T/imunologia , Linfócitos T/patologia
15.
Lipids ; 43(11): 1075-83, 2008 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-18830732

RESUMO

As a transmembrane chemokine, CXCL16 has been detected in various tissues and organs under normal and pathological conditions, it also plays an important role in macrophages/dendritic cells (DC) and T cell interactions and trafficking during inflammation and immune responses. LysoPtdOH, a bioactive lipid mediator has been indicated to regulate DC and epithelial functions during wound healing and inflammation responses. However, the direct link of CXCL16 expression with lysoPtdOH has not been established. Using monocyte-derived macrophages/DC (MoDC), we investigated the roles of lysoPtdOH in CXCL16 production and cell surface presentation. We found that macrophages/MoDC constitutively express and secrete CXCL16, lysoPtdOH significantly enhanced CXCL16 protein production stimulated with lipopolysaccharide (LPS) by more than twofold, which was reflected by increased mRNA transcription by 64-fold. Production of CXCL16 increased by lysoPtdOH and LPS from macrophages was inhibited around 70% by Pertussis toxin (G(i/o) specific inhibitor), exoC3 (Rho specific inhibitor), and pyrrolidine dithiocarbamate (the NF-kappaB-dependent pathway inhibitor) separately. LysoPtdOH treatment increased macrophages' chemotactic activity to activated T cells. The soluble form of CXCL16 produced by macrophages/MoDC was functionally chemoattractive to T cells.


Assuntos
Movimento Celular/fisiologia , Quimiocina CXCL6/biossíntese , Lipopolissacarídeos/farmacologia , Lisofosfolipídeos/farmacologia , Macrófagos/imunologia , Linfócitos T/imunologia , Citometria de Fluxo , Humanos , Transdução de Sinais , Linfócitos T/metabolismo
16.
BMC Cancer ; 8: 178, 2008 Jun 25.
Artigo em Inglês | MEDLINE | ID: mdl-18578857

RESUMO

BACKGROUND: CXCR2 chemokine ligands CXCL1, CXCL5 and CXCL6 were shown to be involved in chemoattraction, inflammatory responses, tumor growth and angiogenesis. Here, we comparatively analyzed their expression profile in resection specimens from patients with colorectal adenoma (CRA) (n = 30) as well as colorectal carcinoma (CRC) (n = 48) and corresponding colorectal liver metastases (CRLM) (n = 16). METHODS: Chemokine expression was assessed by microdissection, quantitative real-time PCR (Q-RT-PCR), the enzyme-linked immunosorbent assay (ELISA) and immunohistochemistry (IHC). RESULTS: In contrast to CXCL6, we demonstrated CXCL1 and CXCL5 mRNA and protein expression to be significantly up-regulated in CRC and CRLM tissue specimens in relation to their matched tumor neighbor tissues. Moreover, both chemokine ligands were demonstrated to be significantly higher expressed in CRC tissues than in CRA tissues thus indicating a progressive increase in the transition from the premalignant condition to the development of the malignant status. Although a comparative analysis of the CXCL1/CXCL5 protein expression profiles in CRC patients revealed that the absolute expression level of CXCL1 was significantly higher in comparison to CXCL5, mRNA- and protein overexpression of CXCL5 in CRC and CRLM tissues was much more pronounced (80- and 60- fold in CRC tissues, respectively) in comparison to CXCL1 (5- and 3.5- fold in CRC tissues, respectively). CONCLUSION: Our results demonstrate a significant association between CXCL1 and CXCL5 expression with CRC and CRLM suggesting for both chemokine ligands a potential role in the progression from CRA to CRC and thus, in the initiation of CRC.


Assuntos
Adenoma/metabolismo , Carcinoma/metabolismo , Quimiocinas CXC/biossíntese , Neoplasias Colorretais/metabolismo , Adenoma/genética , Adenoma/patologia , Adulto , Idoso , Sequência de Aminoácidos , Carcinoma/genética , Carcinoma/patologia , Quimiocina CXCL1/biossíntese , Quimiocina CXCL1/genética , Quimiocina CXCL5/biossíntese , Quimiocina CXCL5/genética , Quimiocina CXCL6/biossíntese , Quimiocina CXCL6/genética , Quimiocinas CXC/genética , Neoplasias Colorretais/genética , Neoplasias Colorretais/patologia , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Imuno-Histoquímica , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/secundário , Masculino , Pessoa de Meia-Idade , RNA Mensageiro/biossíntese , RNA Mensageiro/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Regulação para Cima
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