Association of the MCP-1-2518 A/G Polymorphism and No Association of Its Receptor CCR2-64 V/I Polymorphism with Lupus Nephritis

Objective. To evaluate whether the A/G polymorphism at position 2518 in the regulatory region of the monocyte chemoattractant protein-1 (MCP-1) or the V/I polymorphism at position 64 of the receptor. CCR2, are associated with lupus nephritis (LN) or any clinical characteristics of the disease or with renal survival in a patient population. Methods. We selected 197 patients with lupus nephritis and 220 matched healthy controls for study. MCP-1 and CCR2 genotyping was performed by polymerase chain reaction. Clinical and laboratory data were compiled from patients` charts over followup that ranged from 6 months to 10 years. Results. The GIG genotype of MCP-1 was more common in LN patients (p = 0.019), while the A allele was associated with healthy controls (p = 0.007) as was the V allele of CCR2 (p = 0.046) compared to LN patients. Clinical index measures [SLE Disease Activity Index (SLEDAI)], immunological markers, renal histology, renal function at enrollment, and renal survival were not influenced by these polymorphisms. A less aggressive renal disease, measured by renal SLEDAI index, was associated with the V allele of the CCR2 gene polymorphism. Conclusion. These findings support that MCP-1 2518 GIG is associated with LN but there was no association of this genotype with renal function or renal survival. When studying CCR2 64 V/I polymorphism we showed a positive association of the V allele with healthy controls but no association of the genotype with LN patients. (First Release March 152010; J Rheumatol 2010;37:776-82; doi:10.3899/jrheum.090681)

Febuxostat, an Inhibitor of Xanthine Oxidase, Suppresses Lipopolysaccharide-Induced MCP-1 Production via MAPK Phosphatase-1-Mediated Inactivation of JNK.

Excess reactive oxygen species (ROS) formation can trigger various pathological conditions such as inflammation, in which xanthine oxidase (XO) is one major enzymatic source of ROS. Although XO has been reported to play essential roles in inflammatory conditions, the molecular mechanisms underlying the involvement of XO in inflammatory pathways remain unclear. Febuxostat, a selective and potent inhibitor of XO, effectively inhibits not only the generation of uric acid but also the formation of ROS. In this study, therefore, we examined the effects of febuxostat on lipopolysaccharide (LPS)-mediated inflammatory responses. Here we show that febuxostat suppresses LPS-induced MCP-1 production and mRNA expression via activating MAPK phosphatase-1 (MKP-1) which, in turn, leads to dephosphorylation and inactivation of JNK in macrophages. Moreover, these effects of febuxostat are mediated by inhibiting XO-mediated intracellular ROS production. Taken together, our data suggest that XO mediates LPS-induced phosphorylation of JNK through ROS production and MKP-1 inactivation, leading to MCP-1 production in macrophages. These studies may bring new insights into the novel role of XO in regulating inflammatory process through MAPK phosphatase, and demonstrate the potential use of XO inhibitor in modulating the inflammatory processes.

Genetic polymorphisms of RANTES, IL1-A, MCP-1 and TNF-A genes in patients with prostate cancer

BACKGROUND Inflammation has been implicated as an etiological factor in several human cancers, including prostate cancer. Allelic variants of the genes involved in inflammatory pathways are logical candidates as genetic determinants of prostate cancer risk. The purpose of this study was to investigate whether single nucleotide polymorphisms of genes that lead to increased levels of pro-inflammatory cytokines and chemokines are associated with an increased prostate cancer risk. METHODS A case-control study design was used to test the association between prostate cancer risk and the polymorphisms TNF-A-308 A/G (rs 1800629), RANTES-403 G/A (rs 2107538), IL1-A-889 C/T (rs 1800587) and MCP-1 2518 G/A (rs 1024611) in 296 patients diagnosed with prostate cancer and in 311 healthy controls from the same area. RESULTS Diagnosis of prostate cancer was significantly associated with TNF-A GA + AA genotype (OR, 1.61; 95% CI, 1.09-2.64) and RANTES GA + AA genotype (OR, 1.44; 95% CI, 1.09-2.38). A alleles in TNF-A and RANTES influenced prostate cancer susceptibility and acted independently of each other in these subjects. No epistatic effect was found for the combination of different polymorphisms studied. Finally, no overall association was found between prostate cancer risk and IL1-A or MCP-1 polymorphisms. CONCLUSION Our results and previously published findings on genes associated with innate immunity support the hypothesis that polymorphisms in proinflammatory genes may be important in prostate cancer development.

Profile of circulating levels of IL-1Ra, CXCL10/IP-10, CCL4/MIP-1β and CCL2/MCP-1 in dengue fever and parvovirosis

Dengue virus (DENV) and parvovirus B19 (B19V) infections are acute exanthematic febrile illnesses that are not easily differentiated on clinical grounds and affect the paediatric population. Patients with these acute exanthematic diseases were studied. Fever was more frequent in DENV than in B19V-infected patients. Arthritis/arthralgias with DENV infection were shown to be significantly more frequent in adults than in children. The circulating levels of interleukin (IL)-1 receptor antagonist (Ra), CXCL10/inducible protein-10 (IP-10), CCL4/macrophage inflammatory protein-1 beta and CCL2/monocyte chemotactic protein-1 (MCP-1) were determined by multiplex immunoassay in serum samples obtained from B19V (37) and DENV-infected (36) patients and from healthy individuals (7). Forward stepwise logistic regression analysis revealed that circulating CXCL10/IP-10 tends to be associated with DENV infection and that IL-1Ra was significantly associated with DENV infection. Similar analysis showed that circulating CCL2/MCP-1 tends to be associated with B19V infection. In dengue fever, increased circulating IL-1Ra may exert antipyretic actions in an effort to counteract the already increased concentrations of IL-1β, while CXCL10/IP-10 was confirmed as a strong pro-inflammatory marker. Recruitment of monocytes/macrophages and upregulation of the humoral immune response by CCL2/MCP-1 by B19V may be involved in the persistence of the infection. Children with B19V or DENV infections had levels of these cytokines similar to those of adult patients.

Angiotensin II stimulates MCP-1 production in rat glomerular endothelial cells via NAD(P)H oxidase-dependent nuclear factor-kappa B signaling

Angiotensin II (Ang II) plays a crucial role in the pathogenesis of renal diseases. The objective of the present study was to investigate the possible inflammatory effect of Ang II on glomerular endothelial cells and the underlying mechanism. We isolated and characterized primary cultures of rat glomerular endothelial cells (GECs) and observed that Ang II induced the synthesis of monocyte chemoattractant protein-1 (MCP-1) in GECs as demonstrated by Western blot. Ang II stimulation, at concentrations ranging from 0.1 to 10 µm, of rat GECs induced a rapid increase in the generation of reactive oxygen species as indicated by laser fluoroscopy. The level of p47phox protein, an NAD(P)H oxidase subunit, was also increased by Ang II treatment. These effects of Ang II on GECs were all reduced by diphenyleneiodonium (1.0 µm), an NAD(P)H oxidase inhibitor. Ang II stimulation also promoted the activation of nuclear factor-kappa B (NF-κB). Telmisartan (1.0 µm), an AT1 receptor blocker, blocked all the effects of Ang II on rat GECs. These data suggest that the inhibition of NAD(P)H oxidase-dependent NF-κB signaling reduces the increase in MCP-1 production by GECs induced by Ang II. This may provide a mechanistic basis for the benefits of selective AT1 blockade in dealing with chronic renal disease.

MCP-1 induces migration of adult neural stem cells

As a model for brain inflammation we previously studied transcriptional profiles of tumor necrosis factor-alpha (TNF)treated U373 astroglioma cells. In previous work we were able to demonstrate that the chemokine monocyte chemoattractant protein-1 (MCP-1, SCYA2, CCL2, MCAF) expression in U373 cells was inducible by TNF-alpha treatment. Demonstrably MCP-1 mRNA and protein expression in U373 cells was sustainable over time and at the highest level of all genes analyzed (Schwamborn et al., BMC Genomics 4, 46, 2003). In the hematopoietic system MCP-1 is a CC chemokine that attracts monocytes, memory T lymphocytes, and natural killer cells. In search of further functions in brain inflammation we tested the hypothesis that MCP-1 acts as a chemokine on neural stem cells. Here we report that MCP-1 activates the migration capacity of rat-derived neural stem cells. The migration of stem cells in a Boyden chamber analysis was elevated after stimulation with MCP-1. Time-lapse video microscopy visualized the migration of single stem cells from neurospheres in MCP-1-treated cultures, whereas untreated cultures depicted no migration at all, but showed signs of sprouting. Expression of the MCP-1 receptor CCR2 in neurosphere cultures was verified by RT-PCR and immunofluorescence microscopy. Supernatants from TNF-treated U373 cells also induced migration of neural stem cells.

Expressão das quimiocinas MIP-1α e MCP-1 em relação à carga parasitária em cães com leishmaniose visceral

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Urinary MCP-1 and RBP: Independent predictors of renal outcome in macroalbuminuric diabetic nephropathy

Background: Albuminuria has been considered a sine qua non condition for the diagnosis of diabetic nephropathy (DN) and has been widely used as a surrogate outcome of chronic kidney disease (CKD). However, recent data suggest that albuminuria may fail as a biomarker in a subset of patients, and the search for novel markers is intense. Methods: We analyzed the role of urinary RBP and of serum and urinary cytokines (TGF-beta, MCP-1 and VEGF) as predictors of the risk of dialysis. doubling of serum creatinine or death (primary outcome. PO) in 56 type 2 diabetic patients with macroalbuminuric DN. Results: Mean follow-up time was 30.7 +/- 10 months. Urinary RBP and MCP-1 were significantly higher in patients presenting the PO, whereas no difference was shown for TGF-beta or VEGF. In the Cox regression, urinary RBP. MCP-1 and VEGF were positively associated and serum VEGF was inversely related to the risk of the PO. However, after adjustments for creatinine clearance, proteinuria, and blood pressure only urinary RBP (OR 11.6; 95% CI 2.7-49.2, p = 0.001 for log RBP) and urinary MCP-1 (OR 11.0; 95% CI 1.6-76.4, p = 0.02 for log MCP-1) remained as significant independent predictors of the PO. Conclusion: Urinary RBP and MCP-1 are independently related to the risk of CKD progression in patients with macroalbuminuric DN. Whether these biomarkers have a role in the setting of normoalbuminuria and microalbuminuria in DN should be further investigated. (C) 2012 Elsevier Inc. All rights reserved.

In vivo hydroquinone exposure impairs MCP-1 secretion and monocyte recruitment into the inflamed lung

Alveolar macrophages (AMs) are important cells in the resolution of the inflammatory process and they come into direct contact with inhaled pollutants. Hydroquinone (HQ) is an environmental pollutant and a component of cigarette smoke that causes immunosuppressive effects. In the present work, we showed that mice exposed to low levels of aerosolized HQ (25 ppm; 1 h/day/5 days) presented impaired mononuclear cell migration to the lipopolysaccharide (LPS)-inflamed lung. This may have been due to reduced monocyte chemoattractant protein-1 (MCP-1) secretion into bronchoalveolar lavage fluid (BALF), and it was not related to alterations to mononuclear cell mobilization into the blood or adhesion molecules expression on mononuclear cell membranes. Corroborating the actions of HQ on MCP-1 secretion, reduced MCP-1 concentrations were also found in the supernatant of ex vivo AM and tracheal tissue collected from HQ-exposed mice. A direct action of HQ on MCP-1 secretion, resulting from impaired gene synthesis, was verified by in vitro incubation of naive AMs or tracheal tissue with HQ. The role of reduced levels of MCP-1 in the BALF on monocyte migration was analysed in the human monocytic lineage THP-1 in in vitro chemotaxis assays, which showed that the reduced concentrations of MCP-1 found in the BALF or cell supernatants from HQ-exposed mice impaired cell migration. Considering the fact that MCP-1 presents a broad spectrum of actions on pathophysiological conditions and that resident mononuclear cells are involved in lung tissue homeostasis and in immune host defence, the mechanism of HQ toxicity presented herein might be relevant to the genesis of infectious lung diseases in smokers and in inhabitants of polluted areas. (C) 2012 Elsevier Ireland Ltd. All rights reserved.

Induktion und Signaltransduktion bei der Expression des Chemokins MCP-1 in Monozyten

Das Chemokin 'Monocyte Chemoattractant Protein-1' (MCP-1) spielt bei inflammatorischen Erkrankungen eine wesentliche Rolle. Verschiedene Zelltypen produzieren MCP-1. Es interessierte, welche Stimuli in Monozyten MCP-1 induzieren können und welche Signaltransduktionskaskaden daran beteiligt sind. Darüber hinaus sollte die Rolle einzelner Transkriptionsfaktoren und Promotorregionen des MCP-1-Gens untersucht werden.Komponenten Gram-positiver und -negativer Bakterien, Phorbolester und Substanzen, die die intrazelluläre Calciumkonzentration erhöhen, induzierten die MCP-1-Expression in einer humanen myelomonozytären Zellinie (THP-1) und in frisch isolierten Monozyten. Die mit Lipopolysaccharid (LPS)-induzierte MCP-1-Expression war stark von der MAPK/ERK-Kinase (MEK)-1/-2 und von I-kappaB Kinasen beziehungsweise NF-kappaB abhängig, dagegen scheinen Calcineurin, Calmodulinkinasen und die 'Mitogen-Activated Protein Kinase' p38 keine entscheidende Rolle zu spielen. Die Thapsigargin (TG)-induzierte MCP-1-Bildung durch Erhöhung der intrazellulären Calciumkonzentration war zusätzlich von Calcineurin und Calmodulinkinasen abhängig. Als nukleäre Transkriptionsfaktoren wurden bei der LPS-Stimulation NF-kappaB sowie AP-1 und zusätzlich NF-ATc3 bei Stimulation durch TG nachgewiesen. Die Untersuchung des MCP-1-Promotors konnte eine Bindung von NF-kappaB- und AP-1-Mitglieder an eine bislang nicht untersuchte distale Region und eine AP-1-Bindung an eine proximale Region nachweisen. Die Ergebnisse lassen den Schluß zu, daß die Aktivierung der MCP-1-Expression durch verschiedene Stimuli unter Beteiligung teilweise unterschiedlicher Signaltransduktionswege abläuft und sowohl eine proximale als auch eine distale Promotorregion des MCP-1-Gens daran beteiligt ist.

Sequential cytokine dynamics in chronic rejection of rat renal allografts: roles for cytokines RANTES and MCP-1.

Chronic rejection, the most important cause of long-term graft failure, is thought to result from both alloantigen-dependent and -independent factors. To examine these influences, cytokine dynamics were assessed by semiquantitative competitive reverse transcriptase-PCR and by immunohistology in an established rat model of chronic rejection lf renal allografts. Isograft controls develop morphologic and immunohistologic changes that are similar to renal allograft changes, although quantitatively less intense and at a delayed speed; these are thought to occur secondary to antigen-independent events. Sequential cytokine expression was determined throughout the process. During an early reversible allograft rejection episode, both T-cell associated [interleukin (IL) 2, IL-2 receptor, IL-4, and interferon gamma] and macrophage (IL-1 alpha, tumor necrosis factor alpha, and IL-6) products were up-regulated despite transient immunosuppression. RANTES (regulated upon activation, normal T-cell expressed and secreted) peaked at 2 weeks; intercellular adhesion molecule (ICAM-1) was maximally expressed at 6 weeks. Macrophage products such as monocyte chemoattractant protein (MCP-1) increased dramatically (to 10 times), presaging intense peak macrophage infiltration at 16 weeks. In contrast, in isografts, ICAM-1 peaked at 24 weeks. MCP-1 was maximally expressed at 52 weeks, commensurate with a progressive increase in infiltrating macrophages. Cytokine expression in the spleen of allograft and isograft recipients was insignificant. We conclude that chronic rejection of kidney allografts in rats is predominantly a local macrophage-dependent event with intense up-regulation of macrophage products such as MCP-1, IL-6, and inducible nitric oxide synthase. The cytokine expression in isografts emphasizes the contribution of antigen-independent events. The dynamics of RANTES expression between early and late phases of chronic rejection suggest a key role in mediating the events of the chronic process.

Conservação da qualidade de caqui 'Fuyu' em ambiente refrigerado pela combinação de 1-MCP e atmosfera modificada

O presente estudo analisou os efeitos do inibidor da ação do etileno 1-MCP (1-metilciclopropeno), da AM (atmosfera modificada) e do oxidante de etileno KMnO4 (permanganato de potássio) sobre a qualidade de caqui 'Fuyu' após a armazenagem refrigerada. Os fatores 1-MCP, AM e KMnO4 foram combinados de quatro maneiras, correspondendo aos seguintes tratamentos: T1) Controle + AM + KMnO4;T2) 1-MCP + AM + KMnO4; T3) 1-MCP + AM, e T4) 1-MCP + AA (AA=atmosfera do ar). Frutos maduro-firmes com coloração da casca predominantemente amarela foram colhidos em sete pomares comerciais no nordeste do Estado do Rio Grande do Sul. Parte dos frutos foi exposta a 0.3 µL L-1 de 1-MCP por 12 h em 24 h após a colheita. A seguir, os frutos foram armazenados sob AA ou sob AM induzida por bolsas de polietileno (0,04 mm de espessura), por 20; 40; 60 ou 80 dias a -0,1±0,8ºC. Dois sachês contendo 8,5 g de Alumina-KMnO4foram adicionados em cada uma das bolsas de polietileno dos tratamentos um e dois, antes de elas serem vedadas. Os frutos de cada período de armazenagem refrigerada foram analisados após 0; 3; 6 ou 9 dias de prateleira sob AA a 22±1ºC. O tratamento 1-MCP retardou o amolecimento da polpa, mas não afetou consistentemente o desenvolvimento de 'estrias' e manchas pretas na superfície dos frutos armazenados sob AM contendo KMnO4. A incidência de 'estrias' e manchas pretas em frutos tratados com 1-MCP e armazenados sob AM foi significativamente menor que a de frutos tratados com 1-MCP e armazenados sob AA. Houve efeitos aditivos do 1-MCP e AM na conservação da firmeza e na redução de danos por frio manifestados pela formação de textura gel-firme e manchas translúcidas na casca. O uso de KMnO4 não aumentou a conservação da qualidade dos frutos quando tratados com 1-MCP e armazenados sob AM. O desenvolvimento dos distúrbios da epiderme dependeu do pomar e de períodos de armazenagem e prateleira. No entanto, os benefícios da combinação de 1-MCP e AM sobre a redução desses distúrbios e do amolecimento dos frutos foram consistentes para todos os pomares. Os resultados indicam que a combinação de 1-MCP e AM é um método efetivo para retardar a deterioração de caqui 'Fuyu' durante e após a armazenagem refrigerada.

Resposta da aplicação do 1-MCP em frutos de mamoeiro 'Golden' em diferentes estádios de maturação

O estudo de fatores que influenciam no processo de amadurecimento é fundamental para o planejamento do processo de comercialização, principalmente em frutos com padrão de respiração climatérico e perecível, como é o caso do mamão. Nesse trabalho, avaliou-se o efeito da aplicação do 1-MCP (1-metilciclopropeno) sobre o amadurecimento de frutos de mamoeiro nos estádios 0; 1 e 2 de maturação. O 1-MCP diminui a produção de etileno (≈79%) e a taxa respiratória (≈45%), principalmente em frutos no estádio 0 de maturação. O uso deste inibidor da ação do etileno retardou a perda de coloração verde da casca dos frutos, principalmente em frutos nos estádios 0 e 1 de maturação. Houve redução na perda de firmeza do fruto e do mesocarpo nos estádios 1 e 2. Entretanto, em frutos no estádio 0 de maturação, a firmeza do mesocarpo manteve-se alta, o que pode comprometer a aceitação destes frutos pelo consumidor. O teor de sólidos solúveis não foi influenciado pela aplicação do 1-MCP. O efeito do 1-MCP na redução da atividade das enzimas PME e PG foi maior em frutos nos estádios 0 e 1 de maturação em comparação a frutos no estádio 2 de maturação. A atividade da PME demonstrou crescente aumento ao longo do período de armazenamento, porém a atividade da PG permaneceu baixa ao longo dos cinco primeiros dias, com aumento posterior. Os resultados mostraram que a PME exerce influência significativa na perda de firmeza da polpa nos primeiros dias, com atuação posterior da PG. O 1-MCP mostrou-se eficiente em retardar o processo de amadurecimento de frutos de mamoeiro, tornando-se mais eficiente quando associado a estádios de maturação iniciais.

Alteração da eficiência do 1-MCP com o retardo na sua aplicação após a colheita em maçãs 'Fuji suprema'

Estudos indicam que a eficiência do 1-metilciclopropeno (1-MCP) para conservação pós-colheita de maçãs é máxima quando aplicado até uma semana após a colheita. No entanto, o carregamento das câmaras de armazenagem comerciais com maçãs 'Fuji' pode extender-se por mais de uma semana. Os efeitos da aplicação tardia do 1-MCP para maçãs 'Fuji' não têm sido reportados. Este trabalho teve por objetivo avaliar os efeitos do retardo na aplicação de 1-MCP, a partir da data de colheita, na preservação da firmeza de polpa, acidez titulável (AT) e sólidos solúveis (SS), e na prevenção de escaldadura superficial, escurecimento da polpa e podridões em maçãs 'Fuji Suprema'. Os frutos foram colhidos em pomares comerciais do Estado de Santa Catarina, nas regiões de Fraiburgo (quatro pomares), Bom Retiro (três pomares) e São Joaquim (três pomares), em abril de 2006. Os frutos foram refrigerados 12 h após a colheita e mantidos durante nove meses sob atmosfera do ar a 0,5 ± 0,5 ºC e 90-95% de UR. Os frutos foram tratados com ar (controle) ou 1-MCP (1 µL.L-1, durante 24h), na mesma temperatura de armazenamento, aos 0; 4; 8; 12; 16 ou 20 dias após a colheita. A qualidade dos frutos foi determinada após o armazenamento refrigerado, mais sete dias a 23 ºC. O retardo na aplicação do 1-MCP, por até 20 dias após a colheita, não reduziu sua eficiência na conservação da firmeza da polpa e na prevenção do escurecimento da polpa e podridões, em frutos colhidos nas três regiões, bem como na preservação do teor de SS em frutos colhidos em Bom Retiro e São Joaquim. No entanto, a eficiência do 1-MCP sobre a prevenção da escaldadura superficial foi reduzida quando sua aplicação foi atrasada por 16 ou 20 dias a partir da data de colheita, especialmente nos frutos das regiões que apresentaram maior suscetibilidade ao distúrbio (Fraiburgo e São Joaquim). Os benefícios do 1-MCP sobre a conservação da AT foram reduzidos quando aplicado 20 dias após a colheita nos frutos da região Bom Retiro. Os resultados deste estudo evidenciam os benefícios do tratamento 1-MCP na conservação da qualidade de maçãs 'Fuji Suprema', armazenadas sob atmosfera do ar, por longos perídos, e a redução da eficiência deste tratamento na prevenção da escaldadura superficial, quando a aplicação for atrasada em mais de 12 dias após a colheita dos frutos.