RESUMEN
OBJECTIVES: Chemerin and interleukin (IL)-8 are pro-inflammatory mediators whose role in joint inflammation and cartilage degradation has been demonstrated in in-vitro findings. Studies on their presence in synovial fluid (SF) samples may offer further information on their pathogenic role. The aim of this study was to investigate SF chemerin and IL-8 levels in patients with different joint diseases. METHODS: 37 patients were enrolled: 18 with rheumatoid arthritis (RA), 8 with psoriatic arthritis (PsA) and 11 with osteoarthritis (OA). 41 SF samples were obtained by arthrocentesis in case of knee synovitis. Serum samples were obtained from 13 patients (4 with RA, 6 with PsA and 3 with OA) at the time of arthrocentesis. Chemerin, IL-8, TNF-α and IL-6 levels were measured using commercially available ELISA kits. Immunohistochemical analysis of synovial RA specimens was also performed. RESULTS: No difference in chemerin SF levels emerged between patients with immune-mediated inflammatory arthritides and those with OA (p=0.0656), while subjects with inflammatory arthritis displayed significantly higher levels of SF IL-8 compared to OA (p=0.0020). No significant difference emerged across the three conditions in the serum levels of both chemerin and IL-8. IL-8 strongly correlated with inflammatory markers as ESR, CRP, IL-6 and TNF-α. CONCLUSIONS: We observed similar chemerin SF and serum levels in the three conditions. Although flawed by some limitations, our findings support the emerging concept of OA as an inflammatory disorder. However the increased IL-8 levels we described in patients with inflammatory arthritis suggest a selective involvement of this pro-inflammatory and angiogenic cytokine in these conditions.
Asunto(s)
Artritis Psoriásica/metabolismo , Artritis Reumatoide/metabolismo , Quimiocinas/análisis , Interleucina-8/análisis , Osteoartritis/metabolismo , Líquido Sinovial/metabolismo , Anciano , Artritis Psoriásica/diagnóstico , Artritis Psoriásica/fisiopatología , Artritis Reumatoide/diagnóstico , Artritis Reumatoide/fisiopatología , Biomarcadores/metabolismo , Sedimentación Sanguínea , Proteína C-Reactiva/análisis , Cartílago Articular/metabolismo , Cartílago Articular/fisiopatología , Diagnóstico Diferencial , Femenino , Humanos , Péptidos y Proteínas de Señalización Intercelular , Articulaciones/metabolismo , Articulaciones/fisiopatología , Masculino , Persona de Mediana Edad , Osteoartritis/diagnóstico , Osteoartritis/fisiopatología , Estadística como Asunto , Líquido Sinovial/química , Factor de Necrosis Tumoral alfa/análisisRESUMEN
The beneficial effects of hot springs have been known for centuries and treatments with sulphurous thermal waters are recommended in a number of chronic pathologies as well as acute recurrent infections. However, the positive effects of the therapy are often evaluated in terms of subjective sense of wellbeing and symptomatic clinical improvements. Here, the effects of an S-based compound (NaSH) and of a specific sulphurous thermal water characterized by additional ions such as sodium chloride, bromine and iodine (STW) were investigated in terms of cytokine release and anti-oxidant enzyme activity in primary human monocytes and in saliva from 50 airway disease patients subjected to thermal treatments. In vitro, NaSH efficiently blocked the induction of pro-inflammatory cytokines and counterbalanced the formation of ROS. Despite STW not recapitulating these results, possibly due to the low concentration of S-based compounds reached at the minimum non-toxic dilution, we found that it enhanced the release of IL-10, a potent anti-inflammatory cytokine. Notably, higher levels of IL-10 were also observed in patients' saliva following STW treatment and this increase correlated positively with salivary catalase activity (r2 = 0.19, *p less than 0.01). To our knowledge, these results represent the first evidence suggesting that S-based compounds and STW may prove useful in facing chronic inflammatory and age-related illness due to combined anti-inflammatory and anti-oxidant properties.
Asunto(s)
Antiinflamatorios/farmacología , Antioxidantes/farmacología , Balneología , Enzimas/metabolismo , Manantiales de Aguas Termales , Inflamación/terapia , Interleucina-10/metabolismo , Leucocitos Mononucleares/efectos de los fármacos , Aguas Minerales , Enfermedades Respiratorias/terapia , Adulto , Anciano , Catalasa/metabolismo , Células Cultivadas , Femenino , Glutatión Peroxidasa/metabolismo , Humanos , Inflamación/enzimología , Inflamación/inmunología , Mediadores de Inflamación/metabolismo , Italia , Leucocitos Mononucleares/enzimología , Leucocitos Mononucleares/inmunología , Masculino , Persona de Mediana Edad , NADPH Oxidasas/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Enfermedades Respiratorias/enzimología , Enfermedades Respiratorias/inmunología , Saliva/enzimología , Saliva/inmunología , Sulfuros/farmacología , Superóxido Dismutasa/metabolismo , Factores de TiempoRESUMEN
Molecules representative of different classes of chemotactic agents, including formyl-Met-Leu-Phe (FMLP), C5a, leukotriene B4, platelet-activating factor, and interleukin (IL)-8, caused a rapid reduction in the IL-1 binding capacity by human polymorphonuclear leukocytes (PMN), a cell type expressing predominantly the IL-1 type II decoy receptor (IL-1 decoy RII). N-t-Boc-Met-Leu-Phe, an antagonist for the FMLP receptor, inhibited the loss of IL-1 binding capacity induced by FMLP. Monocyte chemotactic protein 1, a chemokine related to IL-8 but inactive on PMN, had no effect on IL-1 binding in this cell type. FMLP was selected for further detailed analysis of chemoattractant-induced loss of IL-1 binding by PMN. The action of FMLP was rapid, reaching 50% of its maximum (80%) at 30 s, the earliest measurable time point, and plateauing between 10 and 30 min. Dose-response analysis revealed that maximal reduction of IL-1 binding was reached at FMLP concentrations that were also optimal for chemotaxis (50% effective dose = 5 x 10(-9) M). The loss of IL-1 binding capacity caused by FMLP was determined by a reduction in receptor number with no change in their affinity. The effect of FMLP on IL-1 receptor (IL-1R) was selective in that the PMN surface structures IL-8R, CD16, CD18, and major histocompatibility complex class I antigens were unaffected under these conditions. Loss of surface IL-1R was not due to an augumented rate of internalization. FMLP caused rapid release of a 45-kD IL-1-binding molecule identified as the IL-1 decoy RII. After FMLP-induced release, PMN reexpressed newly synthesized receptors, reaching basal levels by 4 h. FMLP-induced release of the IL-1 decoy RII did not impair the responsiveness of PMN to IL-1 in terms of promotion of survival and cytokine production. FMLP-induced release of the IL-1 decoy RII was unaffected by protein synthesis inhibitors, was blocked by certain protease inhibitors, and was mimicked by agents (the Ca++ ionophore A23187 and phorbol myristate acetate) that recapitulate elements in the signal transduction pathway of chemoattractant receptors. The time frame and concentration range of chemoattractant-induced rapid release of the IL-1 decoy RII are consistent with the view that IL-1 decoy RII release is an early event in the multistep process of leukocyte recruitment.(ABSTRACT TRUNCATED AT 400 WORDS)
Asunto(s)
Factores Quimiotácticos/farmacología , Quimiotaxis de Leucocito/fisiología , Neutrófilos/fisiología , Receptores de Interleucina-1/biosíntesis , Secuencia de Aminoácidos , Calcimicina/farmacología , Quimiotaxis de Leucocito/efectos de los fármacos , Complemento C5a/farmacología , Humanos , Técnicas In Vitro , Interleucina-1/metabolismo , Interleucina-1/farmacología , Interleucina-8/farmacología , Leucotrieno B4/farmacología , Datos de Secuencia Molecular , N-Formilmetionina Leucil-Fenilalanina/farmacología , Neutrófilos/efectos de los fármacos , Neutrófilos/inmunología , Oligopéptidos/farmacología , Factor de Activación Plaquetaria/farmacología , Receptores de Interleucina-1/antagonistas & inhibidores , Proteínas Recombinantes/metabolismo , Proteínas Recombinantes/farmacología , Acetato de Tetradecanoilforbol/farmacologíaRESUMEN
A cDNA encoding a novel human chemokine was isolated by random sequencing of cDNA clones from human monocyte-derived macrophages. This protein has been termed macrophage-derived chemokine (MDC) because it appears to be synthesized specifically by cells of the macrophage lineage. MDC has the four-cysteine motif and other highly conserved residues characteristic of CC chemokines, but it shares <35% identity with any of the known chemokines. Recombinant MDC was expressed in Chinese hamster ovary cells and purified by heparin-Sepharose chromatography. NH2-terminal sequencing and mass spectrophotometry were used to verify the NH2 terminus and molecular mass of recombinant MDC (8,081 dalton). In microchamber migration assays, monocyte-derived dendritic cells and IL-2-activated natural killer cells migrated to MDC in a dose-dependent manner, with a maximal chemotactic response at 1 ng/ml. Freshly isolated monocytes also migrated toward MDC, but with a peak response at 100 ng/ml MDC. Northern analyses indicated MDC is highly expressed in macrophages and in monocyte-derived dendritic cells, but not in monocytes, natural killer cells, or several cell lines of epithelial, endothelial, or fibroblast origin. High expression was also detected in normal thymus and less expression in lung and spleen. Unlike most other CC chemokines, MDC is encoded on human chromosome 16. MDC is thus a unique member of the CC chemokine family that may play a fundamental role in the function of dendritic cells, natural killer cells, and monocytes.
Asunto(s)
Quimiocinas CC/genética , Quimiotaxis de Leucocito , Células Dendríticas/fisiología , Células Asesinas Naturales/fisiología , Proteínas Inflamatorias de Macrófagos/genética , Macrófagos/fisiología , Monocitos/fisiología , Secuencia de Aminoácidos , Quimiocina CCL22 , Quimiocinas CC/aislamiento & purificación , Cromosomas Humanos Par 16 , Clonación Molecular , ADN Complementario/genética , Expresión Génica , Humanos , Datos de Secuencia Molecular , Filogenia , ARN Mensajero/genética , Alineación de Secuencia , Homología de Secuencia de Ácido Nucleico , Distribución TisularRESUMEN
The present study was designed to investigate the effect of bacterial lipopolysaccharide (LPS) on C-C chemokine receptors (CCR) expressed in human mononuclear phagocytes. LPS caused a rapid and drastic reduction of CCR2 mRNA levels, which binds MCP-1 and -3. CCR1 and CCR5 mRNAs were also reduced, though to a lesser extent, whereas CXCR2 was unaffected. The rate of nuclear transcription of CCR2 was not affected by LPS, whereas the mRNA half life was reduced from 1.5 h to 45 min. As expected, LPS-induced inhibition of CCR2 mRNA expression was associated with a reduction of both MCP-1 binding and chemotactic responsiveness. The capacity to inhibit CCR2 expression in monocytes was shared by other microbial agents and cytokines (inactivated Streptococci, Propionibacterium acnes, and to a lesser extent, IL-1 and TNF-alpha). In contrast, IL-2 augmented CCR2 expression and MCP-1 itself had no effect. These results suggest that, regulation of receptor expression in addition to agonist production is likely a crucial point in the regulation of the chemokine system.
Asunto(s)
Citocinas , Regulación hacia Abajo , Lipopolisacáridos/farmacología , Monocitos/efectos de los fármacos , Receptores de Quimiocina , Receptores de Citocinas/biosíntesis , Quimiocina CCL2/metabolismo , Quimiocina CCL7 , Quimiotaxis de Leucocito/efectos de los fármacos , Humanos , Proteínas Quimioatrayentes de Monocitos/metabolismo , ARN Mensajero/metabolismo , Receptores CCR2 , Factores de TiempoRESUMEN
T helper cells type 1 (Th1s) that produce interferon-gamma predominantly mediate cellular immune responses and are involved in the development of chronic inflammatory conditions, whereas Th2s which produce large amounts of IL-4 and IL-5 upregulate IgE production and are prominent in the pathogenesis of allergic diseases. The precise factors determining whether Th1- or Th2-mediated immune responses preferentially occur at a peripheral site of antigen exposure are largely unknown. Chemokines, a superfamily of polypeptide mediators, are a key component of the leukocyte recruitment process. Here we report that among four CXC (CXCR1-4) and five CC (CCR1-5) chemokine receptors analyzed, CXCR3 and CCR5 are preferentially expressed in human Th1s. In contrast, Th2s preferentially express CCR4 and, to a lesser extent, CCR3. In agreement with the differential chemokine receptor expression, Th1s and Th2s selectively migrate in response to the corresponding chemokines. The differential expression of chemokine receptors may dictate, to a large extent, the migration and tissue homing of Th1s and Th2s. It may also determine different susceptibility of Th1s and Th2s to human immunodeficiency virus strains using different fusion coreceptors.
Asunto(s)
Quimiotaxis de Leucocito , Receptores de Quimiocina/metabolismo , Células TH1/inmunología , Células Th2/inmunología , Movimiento Celular , Células Clonales , Citocinas/biosíntesis , Sangre Fetal/citología , Sangre Fetal/inmunología , VIH/patogenicidad , Humanos , Inmunidad Celular , Técnicas In Vitro , Receptores del VIH/metabolismo , Receptores Mensajeros de Linfocitos/metabolismo , Células TH1/virología , Células Th2/virologíaRESUMEN
The immunosuppressive and antiinflammatory cytokine interleukin (IL) 10 selectively upregulates the expression of the CC chemokine receptors CCR5, 2, and 1 in human monocytes by prolonging their mRNA half-life. IL-10-stimulated monocytes display an increased number of cell surface receptors for, and better chemotactic responsiveness to, relevant agonists than do control cells. In addition, IL-10-stimulated monocytes are more efficiently infected by HIV BaL. This effect was associated to the enhancement of viral entry through CCR5. These data add support to an emerging paradigm in which pro- and antiinflammatory molecules exert reciprocal and opposing influence on chemokine agonist production and receptor expression.
Asunto(s)
Infecciones por VIH/virología , Interleucina-10/farmacología , Monocitos/virología , Receptores CCR5/metabolismo , Northern Blotting , ADN Viral/metabolismo , Citometría de Flujo , Regulación de la Expresión Génica/genética , Humanos , Cinética , ARN Mensajero/efectos de los fármacos , ARN Mensajero/metabolismo , Receptores CCR1 , Receptores CCR5/genética , Receptores de Quimiocina/genética , Regulación hacia Arriba/efectos de los fármacosRESUMEN
Dendritic cells are potent antigen-presenting cells involved in the initiation of immune responses. The trafficking of these cells to tissues and lymph nodes is mediated by members of the chemokine family. Recently, a novel CC chemokine known as MIP-3alpha or liver and activation-regulated chemokine has been identified from the EMBL/GenBank/DDBJ expressed sequence tag database. In the present study, we have shown that the messenger RNA for MIP-3alpha is expressed predominantly in inflamed and mucosal tissues. MIP-3alpha produced either synthetically or by human embryonic kidney 293 cells is chemotactic for CD34(+)-derived dendritic cells and T cells, but is inactive on monocytes and neutrophils. MIP-3alpha was unable to displace the binding of specific CC or CXC chemokines to stable cell lines expressing their respective high affinity receptors, namely CCR1-5 and CXCR1 and CXCR2, suggesting that MIP-3alpha acts through a novel CC chemokine receptor. Therefore, we used degenerate oligonucleotide-based reverse transcriptase PCR to identify candidate MIP-3alpha receptors in lung dendritic cells. Our results show that the orphan receptor known as GCY-4, CKRL-3, or STRL-22 is a specific receptor for MIP-3alpha, and that its activation leads to pertussis toxin-sensitive and phospholipase C-dependent intracellular Ca2+ mobilization when it is expressed in HEK 293 cells.
Asunto(s)
Quimiocinas CC , Células Dendríticas/inmunología , Pulmón/inmunología , Proteínas Inflamatorias de Macrófagos/metabolismo , Receptores de Quimiocina , Receptores de Citocinas/genética , Secuencia de Aminoácidos , Secuencia de Bases , Unión Competitiva , Calcio/metabolismo , Línea Celular , Quimiocina CCL20 , Clonación Molecular , Cartilla de ADN/genética , Células Dendríticas/metabolismo , Expresión Génica , Humanos , Pulmón/metabolismo , Proteínas Inflamatorias de Macrófagos/genética , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa , ARN Mensajero/genética , ARN Mensajero/metabolismo , Receptores CCR6 , Receptores de Citocinas/metabolismo , Linfocitos T/inmunología , Linfocitos T/metabolismoRESUMEN
Phosphoinositide 3-kinase (PI3K) activity is crucial for leukocyte function, but the roles of the four receptor-activated isoforms are unclear. Mice lacking heterotrimeric guanine nucleotide-binding protein (G protein)-coupled PI3Kgamma were viable and had fully differentiated neutrophils and macrophages. Chemoattractant-stimulated PI3Kgamma-/- neutrophils did not produce phosphatidylinositol 3,4,5-trisphosphate, did not activate protein kinase B, and displayed impaired respiratory burst and motility. Peritoneal PI3Kgamma-null macrophages showed a reduced migration toward a wide range of chemotactic stimuli and a severely defective accumulation in a septic peritonitis model. These results demonstrate that PI3Kgamma is a crucial signaling molecule required for macrophage accumulation in inflammation.
Asunto(s)
Quimiotaxis , Proteínas de Unión al GTP Heterotriméricas/metabolismo , Macrófagos Peritoneales/fisiología , Neutrófilos/fisiología , Peritonitis/inmunología , Fosfatidilinositol 3-Quinasas/metabolismo , Proteínas Serina-Treonina Quinasas , Transducción de Señal , Animales , Factores Quimiotácticos/farmacología , Quimiotaxis de Leucocito/fisiología , Activación Enzimática , Marcación de Gen , Isoenzimas/metabolismo , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Neutrófilos/metabolismo , Peritonitis/enzimología , Peritonitis/patología , Fosfatos de Fosfatidilinositol/metabolismo , Proteínas Proto-Oncogénicas/metabolismo , Proteínas Proto-Oncogénicas c-akt , Estallido RespiratorioRESUMEN
The homeostatic chemokine CXCL13 is preferentially produced in B-follicles and is crucial in the lymphoid organ development by attracting B-lymphocytes that express its selective receptor CXCR5. Follicular dendritic cells (FDCs) have been identified as the main cellular source of this chemokine in lymphoid organs. Recently, genome-wide approaches have suggested follicular CD4 T-helper cells (T(H)F) as additional CXCL13 producers in the germinal centre and the neoplastic counterpart of T(H)F (CD4+ tumour T-cells in angioimmunoblastic T-cell lymphoma) retains the capability of producing this chemokine. In contrast, no data are available on CXCL13 expression on FDC sarcoma (FDC-S) cells. By using multiple approaches, we investigated the expression of CXCL13 at mRNA and protein level in reactive and neoplastic FDCs. In reactive lymph nodes and tonsils, CXCL13 protein is mainly expressed by a subset of FDCs in B-cell follicles. CXCL13 is maintained during FDC transformation, since both dysplastic FDCs from 13 cases of Castleman's disease and neoplastic FDCs from ten cases of FDC-S strongly and diffusely express this chemokine. This observation was confirmed at mRNA level by using RT-PCR and in situ hybridization. Of note, no CXCL13 reactivity was observed in a cohort of epithelial and mesenchymal neoplasms potentially mimicking FDC-S. FDC-S are commonly associated with a dense intratumoural inflammatory infiltrate and immunohistochemistry showed that these lymphocytes express the CXCL13 receptor CXCR5 and are mainly of mantle zone B-cell derivation (IgD+ and TCL1+). In conclusion, this study demonstrates that CXCL13 is produced by dysplastic and neoplastic FDCs and can be instrumental in recruiting intratumoural CXCR5+ lymphocytes. In addition to the potential biological relevance of this expression, the use of reagents directed against CXCL13 can be useful to properly identify the origin of spindle cell and epithelioid neoplasms.
Asunto(s)
Biomarcadores de Tumor/análisis , Quimiocina CXCL13/análisis , Células Dendríticas Foliculares/inmunología , Sarcoma/inmunología , Adolescente , Adulto , Anciano , Enfermedad de Castleman/inmunología , Quimiocina CXCL13/genética , Femenino , Humanos , Inmunohistoquímica , Hibridación in Situ/métodos , Ganglios Linfáticos/inmunología , Masculino , Persona de Mediana Edad , Tonsila Palatina/inmunología , ARN Mensajero/análisis , Receptores CXCR5/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , Linfocitos T/metabolismoRESUMEN
Immunocompromised patients with disseminated human cytomegalovirus (HCMV) infection have circulating PMN carrying HCMV pp65 (antigenemia), infectious virus (viremia), and viral DNA (leukoDNAemia). Because HCMV does not fully replicate in PMN, it is generally hypothesized that virions and viral materials are taken up by phagocytosis from fully permissive HCMV-infected endothelial cells. However, no experimental evidence has ever been provided for these PMN-endothelium interactions. PMN from 11 donors were cocultured with endothelial cells infected with an endothelium-adapted HCMV strain and with human fibroblasts infected with low-passaged clinical and laboratory-adapted HCMV strains. pp65-positive PMN were detected after coculture with both HCMV-infected endothelial and fibroblast cells, provided that wild and not laboratory-adapted strains were used. In addition, cocultured PMN carried infectious virus as demonstrated by virus isolation and presence of complete virus particles by electron microscopy. Moreover, high levels of viral DNA were consistently detected by quantitative PCR in cocultured PMN. Thus, we have generated in vitro the three most important viral parameters detected in patients with disseminated HCMV infection (antigenemia, viremia, and leukoDNAemia). The failure of laboratory-adapted HCMV strain to induce this phenomenon demonstrates that important modifications have occurred in attenuated viral strains affecting basic biological functions.
Asunto(s)
Infecciones por Citomegalovirus/fisiopatología , Infecciones por Citomegalovirus/virología , ADN Viral/fisiología , Endotelio Vascular/virología , Neutrófilos/virología , Fosfoproteínas/inmunología , Proteínas de la Matriz Viral/inmunología , Viremia/virología , Técnicas de Cocultivo , Citomegalovirus/fisiología , Endotelio Vascular/citología , Endotelio Vascular/fisiología , Humanos , Neutrófilos/citología , Neutrófilos/fisiología , Fagocitosis , Replicación ViralRESUMEN
Receptor-mediated agonists, such as FMLP, induce an early, phospholipase D (PLD)-mediated accumulation of phosphatidic acid (PA) which may play a role in the activation of NADPH oxidase in human PMN. We have determined the effect of changes in PA production on O2 consumption in intact PMN and the level of NADPH oxidase activity measured in a cell-free assay. Pretreatment of cells with various concentrations of propranolol enhanced (less than or equal to 200 microM) or inhibited (greater than 300 microM) PLD-induced production of PA (mass and radiolabel) in a manner that correlated with enhancement or inhibition of O2 consumption in PMN stimulated with 1 microM FMLP in the absence of cytochalasin B. The concentration-dependent effects of propranolol on FMLP-induced NADPH oxidase activation was confirmed by direct assay of the enzyme in subcellular fractions. In PA extracted from cells pretreated with 200 microM propranolol before stimulation with 1 microM FMLP, phospholipase A1 (PLA1)-digestion for 90 min, followed by quantitation of residual PA, showed that a minimum of 44% of PA in control (undigested) sample was diacyl-PA; alkylacyl-PA remained undigested by PLA1. Propranolol was also observed to have a concentration-dependent enhancement of mass of 1,2-DG formed in PMN stimulated with FMLP. DG levels reached a maximum at 300 microM propranolol and remained unchanged up to 500 microM propranolol. However, in contrast to PA levels, the level of DG produced did not correlate with NADPH oxidase activation. Exogenously added didecanoyl-PA activated NADPH oxidase in a concentration-dependent manner (1-300 microM) in a reconstitution assay using membrane and cytosolic fractions from unstimulated PMN. In addition, PA synergized with SDS for oxidase activation. Taken together, these results indicate that PA plays a second messenger role in the activation of NADPH oxidase in human PMN and that regulation of phospholipase D is a key step in the activation pathway.
Asunto(s)
NADH NADPH Oxidorreductasas/metabolismo , Neutrófilos/enzimología , Ácidos Fosfatidicos/fisiología , Sistemas de Mensajero Secundario/fisiología , Diglicéridos/metabolismo , Activación Enzimática/efectos de los fármacos , Humanos , N-Formilmetionina Leucil-Fenilalanina/farmacología , NADPH Oxidasas , Fosfolipasa D/metabolismo , Propranolol/farmacologíaRESUMEN
Chemokines may control the macrophage infiltrate found in many solid tumors. In human ovarian cancer, in situ hybridization detected mRNA for the macrophage chemokine monocyte chemoattractant protein-1 (MCP-1) in 16/17 serous carcinomas, 4/4 mucinous carcinomas, 2/2 endometrioid carcinomas, and 1/3 borderline tumors. In serous tumors, mRNA expression mainly localized to the epithelial areas, as did immunoreactive MCP-1 protein. In the other tumors, both stromal and epithelial expression were seen. All tumors contained variable numbers of cells positive for the macrophage marker CD68. MCP-1 mRNA was also detected in the stroma of 5/5 normal ovaries. RT-PCR demonstrated mRNA for MCP-1 in 7/7 serous carcinomas and 6/6 ovarian cancer cell lines. MCP-1 protein was detected by ELISA in ascites from patients with ovarian cancer (mean 4.28 ng/ml) and was produced primarily by the cancer cells. Human MCP-1 protein was also detected in culture supernatants from cell lines and in ascites from human ovarian tumor xenografts which induce a peritoneal monocytosis in nude mice. We conclude that the macrophage chemoattractant MCP-1 is produced by epithelial ovarian cancer and that the tumor cells themselves are probably a major source. MCP-1 may contribute to the accumulation of tumor-associated macrophages, which may subsequently influence tumor behavior.
Asunto(s)
Adenocarcinoma/metabolismo , Adenocarcinoma/patología , Factores Quimiotácticos/análisis , Factores Quimiotácticos/biosíntesis , Citocinas/análisis , Neoplasias Ováricas/metabolismo , Neoplasias Ováricas/patología , Animales , Ascitis , Secuencia de Bases , Línea Celular , Quimiocina CCL2 , Cistadenocarcinoma Seroso/metabolismo , Cistadenocarcinoma Seroso/patología , Cartilla de ADN , Neoplasias Endometriales/metabolismo , Neoplasias Endometriales/patología , Epitelio/metabolismo , Epitelio/patología , Femenino , Expresión Génica , Humanos , Inmunohistoquímica , Hibridación in Situ , Macrófagos/patología , Ratones , Ratones Desnudos , Datos de Secuencia Molecular , Ovario/citología , Ovario/metabolismo , Ovario/patología , Reacción en Cadena de la Polimerasa , ARN Mensajero/biosíntesis , Trasplante HeterólogoRESUMEN
Inflammation is a process to protect the host against infection and danger signals. However, many pathologic conditions, including autoimmune diseases, are sustained by perpetual activation of the inflammatory process. In the past few years our knowledge about the molecular basis of inflammation have been uncovered and now much is known about the primary role of pro-inflammatory cytokines such as IL-1 and TNF. In the early '90s, anti-cytokine therapies started and confirmed the primary role of TNF in autoimmune diseases, such as rheumatoid arthritis, Crohn's Disease and psoriasis. Increasing understanding of the role of inflammatory mediators in inflammation and diseases is opening new avenues for the treatment of inflammatory-based diseases through selective targeting of cytokines and lipid mediators.
Asunto(s)
Anticuerpos Monoclonales/uso terapéutico , Artritis Reumatoide/inmunología , Citocinas/metabolismo , Mediadores de Inflamación/metabolismo , Inflamación/inmunología , Lupus Eritematoso Sistémico/inmunología , Animales , Anticuerpos Monoclonales/inmunología , Artritis Reumatoide/tratamiento farmacológico , Linfocitos B/inmunología , Citocinas/inmunología , Humanos , Mediadores de Inflamación/inmunología , Lupus Eritematoso Sistémico/tratamiento farmacológico , Factor de Necrosis Tumoral alfa/antagonistas & inhibidores , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
The pathomechanism of most autoimmune skin diseases is still elusive; however, recent clinical and basic research is leading novel insights into the cellular and molecular biological underlying pathways. Several types of infectious skin diseases are infiltrated by significant number of gamma/delta T cells and similar observations have been made in selected immune-mediated skin conditions. In particular, a role for gamma/delta T cells has been suggested in discoid lupus erythematosus, contact dermatitis, herpetiformis dermatitis, necrotizing cutaneous vasculitis, and cutaneous lesions of systemic sclerosis. The pathogenesis of these diseases is different and this may suggest multiple potential functions of this subset of T cells in the immune system of the skin. Furthermore, most T cells infiltrating tissue and organs undergoing fibrosis have the potential to produce high levels of interleukin 4. This is particularly true for the CD8+ or CD4+ CD8+ double positive T-cell subsets. Furthermore, leukocyte recruitment is a key event in immunity and a better understanding of the signals involved in autoimmune diseases constitutes a valuable basis for the development of new strategies, which control leukocyte migration and function under pathological conditions.
Asunto(s)
Enfermedades Autoinmunes/inmunología , Inmunidad Celular/inmunología , Enfermedades de la Piel/inmunología , Linfocitos T/inmunología , Apoptosis/inmunología , Enfermedades Autoinmunes/etiología , Fibroblastos/inmunología , Humanos , Esclerodermia Sistémica/inmunología , Enfermedades de la Piel/etiología , Subgrupos de Linfocitos T/inmunologíaRESUMEN
BACKGROUND: Follicular center lymphoma displays widespread lymph node involvement at diagnosis. The chemoattractants that control the locomotion of follicular center lymphoma B cells have not been established. Stromal cell-derived factor-1 (SDF-1) is a CXC-class chemokine that enhances the migration of normal human B cells and is expressed in peripheral lymphoid tissues. Here we have investigated 1) whether SDF-1 stimulates the in vitro locomotion of follicular center lymphoma B cells and of their presumed normal counterparts (i. e., germinal center B cells) and 2) whether the same cells express SDF-1 transcripts. METHODS: B cells were purified by immunomagnetic bead manipulation. Messenger RNA was detected by reverse transcription-polymerase chain reaction. Migration was assessed by the filter and collagen invasion assays. All P values were two sided. RESULTS: Follicular center lymphoma B lymphocytes showed a statistically significant migratory response to 300 ng/mL SDF-1, both in the filter and in the collagen assays (P =.002 for each). Such response was mediated by the SDF-1 receptor, CXCR4. CD40 monoclonal antibody (MAb) and tonsillar germinal center B cells treated with CD40 MAb and recombinant interleukin 4, but not freshly isolated, migrated statistically significantly faster in the presence than in the absence of SDF-1 (P =.002 in both filter and collagen assays). Freshly isolated follicular center lymphoma and germinal center B cells expressed SDF-1 transcripts. CONCLUSIONS: This study shows that SDF-1 substantially enhances the migration of follicular center lymphoma B cells but not the migration of freshly purified germinal center B cells. This difference may be related to the extended survival of follicular center lymphoma versus germinal center B cells. SDF-1 produced in follicular center lymphoma lymph nodes may play a role in the local dissemination of tumor cells.
Asunto(s)
Linfocitos B/fisiología , Quimiocinas CXC/farmacología , Factores Quimiotácticos/farmacología , Linfoma Folicular/metabolismo , Anticuerpos Monoclonales/farmacología , Linfocitos B/metabolismo , Secuencia de Bases , Antígenos CD40/inmunología , Antígenos CD40/fisiología , Quimiocina CXCL12 , Quimiocinas CXC/genética , Quimiotaxis , Expresión Génica , Humanos , Ganglios Linfáticos/metabolismo , Linfoma Folicular/genética , Datos de Secuencia Molecular , Receptores CXCR4/metabolismo , Receptores de Interleucina-4/inmunología , Receptores de Interleucina-4/fisiologíaRESUMEN
NK cells are present mostly in blood and spleen but under certain pathological and physiological conditions rapidly accumulate at extrahematic sites. The present study investigates the responsiveness of NK cells to C-C chemokines and the mechanisms of emigration from the bloodstream. MCP-1 induced migration across polycarbonate filters of IL-2-activated NK cells, whereas it was a weak attractant for unstimulated cells. The related chemokines MCP-2 and MCP-3 were also active. IL-2-activated NK cells showed specific binding sites for labeled MCP-1, and cell migration was inhibited by both cholera and Bordetella pertussis toxins. In agreement with functional assays the expression of mRNA specific for MCP-1 receptors was detectable only in IL-2-activated NK cells. The ability of NK cells to respond to MCP-1 and related chemokines may be one important determinant of NK cell emigration and recruitment in tissues.
RESUMEN
The immunobiology of dendritic cells (DC) involves localization in tissues and trafficking via the lymph or blood to lymphoid organs. Appropriate assays representative of different steps of DC trafficking (e.g., reverse transmigration) provide the tools to dissect the migratory properties of these cells. Chemokines have emerged as important regulators of DC migration. DC are both the target and the source of chemokines. DC express receptors for and respond to a set of chemoattractants that overlap with, but are distinct from, those active on other leukocytes. Differential expression of the CCR6 receptor reveals heterogeneity among DC populations. Functional maturation is associated with loss of responsiveness to chemokines present at sites of inflammation and acquisition of a receptor repertoire that renders these cells responsive to signals that guide their localization in lymphoid organs. A better understanding of the molecular basis of DC trafficking may provide molecular and conceptual tools to direct and modulate DC traffic as a strategy to up-regulate and orient specific immunity.
Asunto(s)
Quimiocinas/fisiología , Células Dendríticas/fisiología , Animales , Moléculas de Adhesión Celular/metabolismo , Movimiento Celular/fisiología , Quimiocinas/biosíntesis , Quimiocinas/inmunología , Células Dendríticas/inmunología , Células Dendríticas/metabolismo , Endotelio Vascular , Regulación de la Expresión Génica , Humanos , Neoplasias/inmunología , Receptores de Quimiocina/metabolismoRESUMEN
Macrophage-derived chemokine (MDC) is a CC chemokine paradigmatic of emerging aspects of chemokine immunobiology. It is constitutively expressed, yet microbial products and cytokines regulate its expression with divergent effects of type II (IL-4 and IL-13) and type I (interferon) cytokines. Processing of the mature protein by dipeptidyl peptidase IV/CD26 provides a further level of regulation. It acts on diverse cellular targets including dendritic cells (DC), NK cells, and T cell subsets. Among these, MDC is a potent attractant for CCR4 expressing polarized Th2 and Tc2 cells, and evidence is consistent with a role of this chemokine as an amplification loop of polarized type II responses. Emerging indications on the involvement of MDC in diverse pathologies, ranging from allergic reactions to HIV infection and neoplasia, are discussed.
Asunto(s)
Quimiocinas CC/inmunología , Quimiocina CCL22 , Quimiocinas CC/metabolismo , Humanos , Macrófagos/inmunología , Macrófagos/metabolismoRESUMEN
The recruitment of T cells into the skin is regulated by chemokines released by resident cells. In this study, we found that normal human keratinocytes activated with Th1-derived supernatant (sup) expressed early (6-12 h) IP-10/CXCL10, MCP-1/CCL2, IL-8/CXCL8, and I-309/CCL1 mRNAs and with slower kinetics (24-96 h), RANTES/CCL5 and MDC/CCL22 mRNAs. Upon stimulation with the Th1 sup, keratinocytes secreted high levels of RANTES, IP-10, MCP-1, and IL-8 and moderate levels of I-309 and MDC. Although much less efficiently, Th2 sup could also induce keratinocyte expression of IL-8, IP-10, RANTES, and MCP-1 but not of I-309 and MDC. TARC/CCL17 was not significantly induced by any stimuli. Sup from keratinocytes activated with Th1-derived cytokines elicited a strong migratory response of Th1 cells and a limited migration of Th2 cells, whereas sup from Th2-activated keratinocytes promoted a moderate migration of Th1 and Th2 lymphocytes. Thus, keratinocytes appear considerably more sensitive to Th1- than to Th2-derived lymphokines in terms of chemokine release and can support the preferential accumulation of Th1 lymphocytes in the skin.