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1.
Pancreatology ; 19(3): 401-408, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30833212

RESUMO

OBJECTIVE: Chemerin, an adipokine, works as the chemoattractant for the immune cells. The role of chemerin in the inflammatory reaction is controversial. Chemerin has been shown to aggravate the inflammatory response, but other studies demonstrated its anti-inflammatory influence. This study assessed the effects of chemerin on acute pancreatitis (AP) in vivo and in vitro. METHODS: For in vivo experiments male Wistar rats were used. For in vitro study rat pancreatic AR42J cells were employed. Chemerin (1, 5 or 10 µg/kg) was given to the rats prior to the induction of AP by subcutaneous caerulein infusion (25 µg/kg). For in vitro studies cells were subjected to caerulein (10 nM) with or without chemerin (100 nM). Serum amylase activity was measured by enzymatic method, serum TNFα concentration - by ELISA kit. Western-blot was used to examine cellular proteins. RESULTS: AP was confirmed by histological examination. Chemerin given to AP rats decreased histological manifestations of AP, reduced serum amylase activity and TNFα concentration. In AR42J cells subjected to caerulein with addition of chemerin signal for TNFα was reduced comparing to the cultures treated with caerulein alone. Analysis of the dynamics of nuclear translocation for p50, p65 and Bcl-3 points out to NF-κB attenuation as a mechanism of observed anti-inflammatory action of chemerin. CONCLUSION: Chemerin significantly alleviated severity of AP in the rat, this is possibly due to the inhibition of pro-inflammatory signaling in the pancreatic cells.


Assuntos
Quimiocinas/uso terapêutico , Peptídeos e Proteínas de Sinalização Intercelular/uso terapêutico , NF-kappa B/metabolismo , Pancreatite/induzido quimicamente , Animais , Linhagem Celular , Ceruletídeo/toxicidade , Quimiocinas/administração & dosagem , Relação Dose-Resposta a Droga , Regulação da Expressão Gênica/efeitos dos fármacos , Peptídeos e Proteínas de Sinalização Intercelular/administração & dosagem , Masculino , Pâncreas/citologia , Pancreatite/tratamento farmacológico , Ratos , Ratos Wistar
2.
Br J Cancer ; 118(10): 1337-1348, 2018 05.
Artigo em Inglês | MEDLINE | ID: mdl-29717200

RESUMO

BACKGROUND: Chemerin, a known chemoattractant, participates in multiple biological events. However, its role in cancer remains largely unknown. METHODS: Chemerin expression was evaluated by real-time PCR, western blot and immunohistochemistry. Forced expression, RNAi, immunoprecipitation, etc. were used in function and mechanism study. Mouse models of extrahepatic and intrahepatic metastasis were employed to evaluate the therapeutic potential of chemerin. RESULTS: Chemerin expression was significantly downregulated in hepatocellular carcinoma, and associated with poor prognosis of HCC patients. Forced expression of chemerin inhibited in vitro migration, invasion and in vivo metastasis of HCC cells. Administration of chemerin effectively suppressed extrahepatic and intrahepatic metastases of HCC cells, resulting in prolonged survival of tumour-bearing nude mice. Chemerin upregulated expression and phosphatase activity of PTEN by interfering with PTEN-CMKLR1 interaction, leading to weakened ubiquitination of PTEN and decreased p-Akt (Ser473) level, which was responsible for suppressed migration, invasion and metastasis of HCC cells. Positive correlation between chemerin and PTEN, and reverse correlation between chemerin and p-Akt (Ser473) were also observed in HCC clinical samples and intrahepatic mouse model in vivo. CONCLUSIONS: Our study has revealed the suppressive role and therapeutic potential of chemerin in HCC metastasis, providing both a prognostic marker and drug candidate for HCC.


Assuntos
Carcinoma Hepatocelular/tratamento farmacológico , Quimiocinas/administração & dosagem , Peptídeos e Proteínas de Sinalização Intercelular/administração & dosagem , Neoplasias Hepáticas/tratamento farmacológico , PTEN Fosfo-Hidrolase/genética , Receptores de Quimiocinas/genética , Animais , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/patologia , Movimento Celular , Proliferação de Células , Quimiocinas/genética , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Células Hep G2 , Humanos , Peptídeos e Proteínas de Sinalização Intercelular/genética , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/patologia , Camundongos , Invasividade Neoplásica/genética , Invasividade Neoplásica/patologia , Metástase Neoplásica , Proteína Oncogênica v-akt/genética , Transdução de Sinais/efeitos dos fármacos , Ensaios Antitumorais Modelo de Xenoenxerto
3.
Int J Mol Sci ; 18(1)2017 Jan 13.
Artigo em Inglês | MEDLINE | ID: mdl-28098795

RESUMO

Angiogenesis is involved in the inflammation and proliferation stages of wound healing, to bring inflammatory cells to the wound and provide a microvascular network to maintain new tissue formation. An excess of inflammation, however, leads to prolonged wound healing and scar formation, often resulting in unfavourable outcomes such as amputation. CC-chemokines play key roles in the promotion of inflammation and inflammatory-driven angiogenesis. Therefore, inhibition of the CC-chemokine class may improve wound healing. We aimed to determine if the broad-spectrum CC-chemokine inhibitor "35K" could accelerate wound healing in vivo in mice. In a murine wound healing model, 35K protein or phosphate buffered saline (PBS, control) were added topically daily to wounds. Cohorts of mice were assessed in the early stages (four days post-wounding) and in the later stages of wound repair (10 and 21 days post-wounding). Topical application of the 35K protein inhibited CC-chemokine expression (CCL5, CCL2) in wounds and caused enhanced blood flow recovery and wound closure in early-mid stage wounds. In addition, 35K promoted neovascularisation in the early stages of wound repair. Furthermore, 35K treated wounds had significantly lower expression of the p65 subunit of NF-κB, a key inflammatory transcription factor, and augmented wound expression of the pro-angiogenic and pro-repair cytokine TGF-ß. These findings show that broad-spectrum CC-chemokine inhibition may be beneficial for the promotion of wound healing.


Assuntos
Quimiocinas/farmacologia , Neovascularização Fisiológica/efeitos dos fármacos , Cicatrização/efeitos dos fármacos , Administração Tópica , Animais , Biomarcadores/metabolismo , Quimiocinas/administração & dosagem , Colágeno/metabolismo , Inflamação/metabolismo , Inflamação/patologia , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Camundongos Endogâmicos C57BL , Perfusão , Fator de Crescimento Transformador beta/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismo
4.
Vascul Pharmacol ; 88: 30-41, 2017 01.
Artigo em Inglês | MEDLINE | ID: mdl-27890480

RESUMO

Chemerin is an adipokine associated with increased blood pressure, and may link obesity with hypertension. We tested the hypothesis that chemerin-induced contraction of the vasculature occurs via calcium flux in smooth muscle cells. Isometric contraction of rat aortic rings was performed in parallel with calcium kinetics of rat aortic smooth muscle cells to assess the possible signaling pathway. Chemerin-9 (nonapeptide of the chemerin S157 isoform) caused a concentration-dependent contraction of isolated aorta (EC50 100nM) and elicited a concentration-dependent intracellular calcium response (EC50 10nM). Pertussis toxin (Gi inhibitor), verapamil (L-type Ca2+ channel inhibitor), PP1 (Src inhibitor), and Y27632 (Rho kinase inhibitor) reduced both calcium influx and isometric contraction to chemerin-9 but PD098059 (Erk MAPK inhibitor) and U73122 (PLC inhibitor) had little to no effect on either measure of chemerin signaling. Although our primary aim was to examine chemerin signaling, we also highlight differences in the mechanisms of chemerin-9 and recombinant chemerin S157. These data support a chemerin-induced contractile mechanism in vascular smooth muscle that functions through Gi proteins to activate L-type Ca2+ channels, Src, and Rho kinase. There is mounting evidence linking chemerin to hypertension and this mechanism brings us closer to targeting chemerin as a form of therapy.


Assuntos
Aorta/metabolismo , Cálcio/metabolismo , Quimiocinas/metabolismo , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Miócitos de Músculo Liso/metabolismo , Animais , Aorta/efeitos dos fármacos , Canais de Cálcio Tipo L/metabolismo , Quimiocinas/administração & dosagem , Relação Dose-Resposta a Droga , Subunidades alfa Gi-Go de Proteínas de Ligação ao GTP/metabolismo , Hipertensão/fisiopatologia , Peptídeos e Proteínas de Sinalização Intercelular/administração & dosagem , Masculino , Contração Muscular/efeitos dos fármacos , Músculo Liso Vascular/efeitos dos fármacos , Músculo Liso Vascular/metabolismo , Miócitos de Músculo Liso/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Transdução de Sinais/efeitos dos fármacos , Quinases Associadas a rho/metabolismo
5.
Nat Commun ; 7: 12528, 2016 08 19.
Artigo em Inglês | MEDLINE | ID: mdl-27538380

RESUMO

Chemotherapy remains a mainstay of cancer treatment but its use is often limited by the development of adverse reactions. Severe loss of body weight (cachexia) is a frequent cause of death in cancer patients and is exacerbated by chemotherapy. We show that genetic inactivation of vascular endothelial growth factor (VEGF)-A in myeloid cells prevents chemotherapy-induced cachexia by inhibiting skeletal muscle loss and the lipolysis of white adipose tissue. It also improves clearance of senescent tumour cells by natural killer cells and inhibits tumour regrowth after chemotherapy. The effects depend on the chemoattractant chemerin, which is released by the tumour endothelium in response to chemotherapy. The findings define chemerin as a critical mediator of the immune response, as well as an important inhibitor of cancer cachexia. Targeting myeloid cell-derived VEGF signalling should impede the lipolysis and weight loss that is frequently associated with chemotherapy, thereby substantially improving the therapeutic outcome.


Assuntos
Antineoplásicos/uso terapêutico , Caquexia/tratamento farmacológico , Células Matadoras Naturais/imunologia , Células Mieloides/imunologia , Neoplasias/tratamento farmacológico , Fator A de Crescimento do Endotélio Vascular/antagonistas & inibidores , Tecido Adiposo Branco/efeitos dos fármacos , Tecido Adiposo Branco/metabolismo , Animais , Antineoplásicos/farmacologia , Caquexia/etiologia , Caquexia/imunologia , Caquexia/patologia , Quimiocinas/administração & dosagem , Quimiocinas/imunologia , Humanos , Peptídeos e Proteínas de Sinalização Intercelular/administração & dosagem , Peptídeos e Proteínas de Sinalização Intercelular/imunologia , Lipólise/efeitos dos fármacos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Músculo Esquelético/efeitos dos fármacos , Músculo Esquelético/patologia , Células Mieloides/efeitos dos fármacos , Células Mieloides/metabolismo , Neoplasias/complicações , Neoplasias/imunologia , Neoplasias/patologia , Resultado do Tratamento , Fator A de Crescimento do Endotélio Vascular/genética , Fator A de Crescimento do Endotélio Vascular/metabolismo , Ensaios Antitumorais Modelo de Xenoenxerto
6.
Sci Rep ; 6: 26830, 2016 05 26.
Artigo em Inglês | MEDLINE | ID: mdl-27225311

RESUMO

Long-term and reversible changes in body weight are typical of seasonal animals. Thyroid hormone (TH) and retinoic acid (RA) within the tanycytes and ependymal cells of the hypothalamus have been implicated in the photoperiodic response. We investigated signalling downstream of RA and how this links to the control of body weight and food intake in photoperiodic F344 rats. Chemerin, an inflammatory chemokine, with a known role in energy metabolism, was identified as a target of RA. Gene expression of chemerin (Rarres2) and its receptors were localised within the tanycytes and ependymal cells, with higher expression under long (LD) versus short (SD) photoperiod, pointing to a physiological role. The SD to LD transition (increased food intake) was mimicked by 2 weeks of ICV infusion of chemerin into rats. Chemerin also increased expression of the cytoskeletal protein vimentin, implicating hypothalamic remodelling in this response. By contrast, acute ICV bolus injection of chemerin on a 12 h:12 h photoperiod inhibited food intake and decreased body weight with associated changes in hypothalamic neuropeptides involved in growth and feeding after 24 hr. We describe the hypothalamic ventricular zone as a key site of neuroendocrine regulation, where the inflammatory signal, chemerin, links TH and RA signaling to hypothalamic remodeling.


Assuntos
Quimiocinas/fisiologia , Metabolismo Energético/fisiologia , Hipotálamo/metabolismo , Peptídeos e Proteínas de Sinalização Intercelular/fisiologia , Fotoperíodo , Animais , Peso Corporal/efeitos dos fármacos , Quimiocinas/administração & dosagem , Quimiocinas/farmacologia , Ingestão de Alimentos/efeitos dos fármacos , Metabolismo Energético/efeitos dos fármacos , Epêndima/citologia , Epêndima/metabolismo , Células Ependimogliais/metabolismo , Humanos , Hipotálamo/citologia , Hipotálamo/efeitos dos fármacos , Injeções Intraventriculares , Peptídeos e Proteínas de Sinalização Intercelular/administração & dosagem , Peptídeos e Proteínas de Sinalização Intercelular/farmacologia , Masculino , Plasticidade Neuronal/efeitos dos fármacos , Distribuição Aleatória , Ratos , Ratos Endogâmicos F344 , Ratos Sprague-Dawley , Receptores de Quimiocinas/análise , Receptores de Quimiocinas/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , Hormônios Tireóideos/fisiologia
7.
Nanomedicine (Lond) ; 11(6): 693-714, 2016 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-27003586

RESUMO

Cardiovascular diseases make up one of the main causes of death today, with myocardial infarction and ischemic heart disease contributing a large share of the deaths reported. With mainstream clinical therapy focusing on palliative medicine following myocardial infarction, the structural changes that occur in the diseased heart will eventually lead to end-stage heart failure. Heart transplantation remains the only gold standard of cure but a shortage in donor organs pose a major problem that led to clinicians and researchers looking into alternative strategies for cardiac repair. This review will examine some alternative methods of treatment using chemokines and drugs carried by nanoparticles as drug delivering agents for the purposes of treating myocardial infarction through the promotion of revascularization. We will also provide an overview of existing studies involving such nanoparticulate drug delivery systems, their reported efficacy and the challenges facing their translation into ubiquitous clinical use.


Assuntos
Sistemas de Liberação de Medicamentos/métodos , Coração/efeitos dos fármacos , Infarto do Miocárdio/tratamento farmacológico , Nanopartículas/química , Angiopoietinas/administração & dosagem , Angiopoietinas/uso terapêutico , Animais , Quimiocinas/administração & dosagem , Quimiocinas/uso terapêutico , Fatores de Crescimento de Fibroblastos/administração & dosagem , Fatores de Crescimento de Fibroblastos/uso terapêutico , Humanos , Neovascularização Fisiológica/efeitos dos fármacos , Fator A de Crescimento do Endotélio Vascular/administração & dosagem , Fator A de Crescimento do Endotélio Vascular/uso terapêutico
8.
Mucosal Immunol ; 9(1): 13-23, 2016 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-25943275

RESUMO

There have been encouraging results for the development of an effective HIV vaccine. However, many questions remain regarding the quality of immune responses and the role of mucosal antibodies. We addressed some of these issues by using a simian immunodeficiency virus (SIV) DNA vaccine adjuvanted with plasmid-expressed mucosal chemokines combined with an intravaginal SIV challenge in rhesus macaque (RhM) model. We previously reported on the ability of CCR9 and CCR10 ligand (L) adjuvants to enhance mucosal and systemic IgA and IgG responses in small animals. In this study, RhMs were intramuscularly immunized five times with either DNA or DNA plus chemokine adjuvant delivered by electroporation followed by challenge with SIVsmE660. Sixty-eight percent of all vaccinated animals (P<0.01) remained either uninfected or had aborted infection compared with only 14% in the vaccine naïve group. The highest protection was observed in the CCR10L chemokines group, where six of nine animals had aborted infection and two remained uninfected, leading to 89% protection (P<0.001). The induction of mucosal SIV-specific antibodies and neutralization titers correlated with trends in protection. These results indicate the need to further investigate the contribution of chemokine adjuvants to modulate immune responses and the role of mucosal antibodies in SIV/HIV protection.


Assuntos
Vacinas contra a AIDS/administração & dosagem , Anticorpos Antivirais/biossíntese , Quimiocinas/imunologia , Imunidade nas Mucosas/efeitos dos fármacos , Síndrome de Imunodeficiência Adquirida dos Símios/prevenção & controle , Vacinas de DNA/administração & dosagem , Vacinas contra a AIDS/genética , Vacinas contra a AIDS/imunologia , Adjuvantes Imunológicos/administração & dosagem , Animais , Quimiocinas/administração & dosagem , Quimiocinas/genética , Feminino , Imunidade Celular/efeitos dos fármacos , Ligantes , Macaca mulatta , Plasmídeos/química , Plasmídeos/imunologia , Receptores CCR/genética , Receptores CCR/imunologia , Receptores CCR10/genética , Receptores CCR10/imunologia , Síndrome de Imunodeficiência Adquirida dos Símios/imunologia , Síndrome de Imunodeficiência Adquirida dos Símios/virologia , Vírus da Imunodeficiência Símia/efeitos dos fármacos , Vírus da Imunodeficiência Símia/imunologia , Vacinação , Vacinas de DNA/genética , Vacinas de DNA/imunologia , Vagina/efeitos dos fármacos , Vagina/imunologia , Vagina/virologia
9.
World J Gastroenterol ; 21(14): 4184-94, 2015 Apr 14.
Artigo em Inglês | MEDLINE | ID: mdl-25892868

RESUMO

AIM: To test whether hepatic stellate cells (HSCs) at different activation stages play different roles in acetaminophen (APAP)-induced acute liver injury (ALI). METHODS: HSCs were isolated from mouse liver and cultured in vitro. Morphological changes of initiation HSCs [HSCs (5d)] and perpetuation HSCs [HSCs (p3)] were observed by immunofluorescence and transmission electron microscopy. The protective effects of HSC-derived molecules, cell lysates and HSC-conditioned medium (HSC-CM) were tested in vivo by survival and histopathological analyses. Liver injury was determined by measuring aminotransferase levels in the serum and by histologic examination of tissue sections under a light microscope. Additionally, to determine the molecular mediators of the observed protective effects of initiation HSCs, we examined HSC-CM using a high-density protein array. RESULTS: HSCs (5d) and HSCs (p3) had different morphological and phenotypic traits. HSCs (5d) presented a star-shaped appearance with expressing α-SMA at non-uniform levels between cells. However, HSCs (p3) evolved into myofibroblast-like cells without lipid droplets and expressed a uniform and higher level of α-SMA. HSC-CM (5d), but not HSC-CM (p3), provided a significant survival benefit and showed a dramatic reduction of hepatocellular necrosis and panlobular leukocyte infiltrates in mice exposed to APAP. However, this protective effect was abrogated at higher cell masses, indicating a therapeutic window of effectiveness. Furthermore, the protein array screen revealed that HSC-CM (5d) was composed of many chemokines and growth factors that correlated with inflammatory inhibition and therapeutic activity. When compared with HSC-CM (p3), higher levels of monocyte chemoattractant protein-1, macrophage inflammatory protein-1γ, hepatocyte growth factor, interleukin-10, and matrix metalloproteinase-2, but lower levels of stem cell factor and Fas-Ligand were observed in HSC-CM (5d). CONCLUSION: These data indicated that initiation HSCs and perpetuation HSCs were different in morphology and protein expression, and provided the first experimental evidence of the potential medical value of initiation HSC-derived molecules in the treatment of ALI.


Assuntos
Doença Hepática Induzida por Substâncias e Drogas/metabolismo , Quimiocinas/metabolismo , Células Estreladas do Fígado/metabolismo , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Fígado/metabolismo , Comunicação Parácrina , Acetaminofen , Animais , Anti-Inflamatórios/administração & dosagem , Forma Celular , Células Cultivadas , Doença Hepática Induzida por Substâncias e Drogas/tratamento farmacológico , Doença Hepática Induzida por Substâncias e Drogas/etiologia , Doença Hepática Induzida por Substâncias e Drogas/patologia , Quimiocinas/administração & dosagem , Meios de Cultivo Condicionados/metabolismo , Modelos Animais de Doenças , Células Estreladas do Fígado/efeitos dos fármacos , Células Estreladas do Fígado/ultraestrutura , Peptídeos e Proteínas de Sinalização Intercelular/administração & dosagem , Fígado/efeitos dos fármacos , Fígado/ultraestrutura , Masculino , Camundongos Endogâmicos C57BL , Necrose , Fenótipo , Transdução de Sinais , Fatores de Tempo
10.
Gen Comp Endocrinol ; 196: 34-40, 2014 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-24287340

RESUMO

The effects of intraperitoneal injections of cholecystokinin (CCK), apelin, ghrelin, and orexin on food intake were examined in the blind cavefish Astyanax fasciatus mexicanus. CCK (50ng/g) induced a decrease in food intake whereas apelin (100ng/g), orexin (100ng/g), and ghrelin (100ng/g) induced an increase in food intake as compared to saline-injected control fish. In order to better understand the central mechanism by which these hormones act, we examined the effects of injections on the brain mRNA expression of two metabolic enzymes, tyrosine hydroxylase (TH), and mechanistic target of rapamycin (mTOR), and of appetite-regulating peptides, CCK, orexin, apelin and cocaine and amphetamine regulated transcript (CART). CCK injections induced a decrease in brain apelin injections, apelin injections induced an increase in TH, mTOR, and orexin brain expressions, orexin treatment increased brain TH expression and ghrelin injections induced an increase in mTOR and orexin brain expressions. CART expression was not affected by any of the injection treatments. Our results suggest that the enzymes TH and mTOR and the hormones CCK, apelin, orexin, and ghrelin all regulate food intake in cavefish through a complex network of interactions.


Assuntos
Apetite/fisiologia , Encéfalo/efeitos dos fármacos , Colecistocinina/administração & dosagem , Ingestão de Alimentos/efeitos dos fármacos , Grelina/administração & dosagem , Hormônios/farmacologia , Peptídeos e Proteínas de Sinalização Intracelular/administração & dosagem , Neuropeptídeos/administração & dosagem , Sirolimo/farmacologia , Tirosina 3-Mono-Oxigenase/metabolismo , Animais , Encéfalo/enzimologia , Characidae , Quimiocinas/administração & dosagem , Colagogos e Coleréticos/administração & dosagem , Imunossupressores/farmacologia , Neurotransmissores/administração & dosagem , Orexinas , RNA Mensageiro/genética , Reação em Cadeia da Polimerase em Tempo Real , Tirosina 3-Mono-Oxigenase/antagonistas & inibidores , Tirosina 3-Mono-Oxigenase/genética
11.
CNS Neurol Disord Drug Targets ; 12(6): 815-29, 2013 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-24047524

RESUMO

Chemokines may play a role in leukocyte migration across the blood-brain barrier (BBB) during neuroinflammation and other neuropathological processes, such as epilepsy. The CC chemokine receptor 5 (CCR5) is a member of CC-chemokine receptor family that binds several chemokines, including CCL3 (macrophage inflammatory protein-1alpha, MIP-1alpha), CCL4 (macrophage inflammatory protein-1beta, MIP-1beta) and CCL5 (RANTES). The current review examines the relationship between CCR5 and the microglia in different neurological disorders and models of CNS injury. CCR5 expression is upregulated in different neurological diseases, where it is often immunolocalized in microglial cells. A multistep cascade couples CCR5 activation by chemokines to Ca(2+) increases in human microglia. Because changes in [Ca(2+)] (i) affect chemotaxis, secretion, and gene expression, pharmacologic modulation of this pathway may alter inflammatory and degenerative processes in the CNS. Consequently, targeting CCR5 by using CCR5 antagonists may attenuate critical aspects of neuroinflammation in different models of neurological disorders. To illustrate the interaction between CCR5 and microglia in the CNS, we used a model of excitotoxicity, and demonstrate the intimate involvement of CCR5 in neuron injury and inflammation attendant to kainic acid (KA)-induced neurotoxicity. CCR5 participates in neuronal injury caused by the excitotoxin, KA, brings inflammatory cells to the sites of KA-induced CNS injury, defines the extent of tissue loss after KA exposure and limits reparative responses. We used a SV40-derived vector carrying an interfering RNA (RNAi) that targets CCR5. Delivered directly to the bone marrow, this vector decreased CCR5 expression in circulating cells. Animals so treated showed greatly reduced expression of CCR5 and its ligands (MIP-1alpha and RANTES) in the CNS, including in the brain vasculature, decreased BBB leakage, demonstrated greater KA-stimulated neurogenesis and increased migration of bone marrow-derived cells to the brain to become neurons. Thus, therapeutic targeting of CCR5 may allow control of potentially injurious neuroinflammatory responses, including decrease in microglial cells activation and proliferation, and facilitate neurogenic repair in seizure-induced and, potentially, other forms of CNS injury.


Assuntos
Modelos Animais de Doenças , Sistemas de Liberação de Medicamentos/métodos , Epilepsia/metabolismo , Inflamação/metabolismo , Inflamação/patologia , Microglia/metabolismo , Regeneração Nervosa/fisiologia , Receptores CCR5/metabolismo , Animais , Antagonistas dos Receptores CCR5 , Morte Celular/efeitos dos fármacos , Morte Celular/fisiologia , Quimiocinas/administração & dosagem , Quimiocinas/metabolismo , Epilepsia/tratamento farmacológico , Epilepsia/patologia , Humanos , Inflamação/tratamento farmacológico , Microglia/efeitos dos fármacos , Regeneração Nervosa/efeitos dos fármacos , Doenças do Sistema Nervoso/tratamento farmacológico , Doenças do Sistema Nervoso/metabolismo , Doenças do Sistema Nervoso/patologia , Neurônios/efeitos dos fármacos , Neurônios/metabolismo , Neurônios/patologia
12.
Blood ; 120(7): 1449-57, 2012 Aug 16.
Artigo em Inglês | MEDLINE | ID: mdl-22618707

RESUMO

The chemokine CCL3/MIP-1α is a risk factor in the outcome of multiple myeloma (MM), particularly in the development of osteolytic bone disease. This chemokine, highly overexpressed by MM cells, can signal mainly through 2 receptors, CCR1 and CCR5, only 1 of which (CCR1) is responsive to CCL3 in human and mouse osteoclast precursors. CCR1 activation leads to the formation of osteolytic lesions and facilitates tumor growth. Here we show that formation of mature osteoclasts is blocked by the highly potent and selective CCR1 antagonist CCX721, an analog of the clinical compound CCX354. We also show that doses of CCX721 selected to completely inhibit CCR1 produce a profound decrease in tumor burden and osteolytic damage in the murine 5TGM1 model of MM bone disease. Similar effects were observed when the antagonist was used prophylactically or therapeutically, with comparable efficacy to that of zoledronic acid. 5TGM1 cells were shown to express minimal levels of CCR1 while secreting high levels of CCL3, suggesting that the therapeutic effects of CCX721 result from CCR1 inhibition on non-MM cells, most likely osteoclasts and osteoclast precursors. These results provide a strong rationale for further development of CCR1 antagonists for the treatment of MM and associated osteolytic bone disease.


Assuntos
Quimiocinas/farmacologia , Quimiocinas/uso terapêutico , Mieloma Múltiplo/tratamento farmacológico , Osteólise/tratamento farmacológico , Receptores CCR1/antagonistas & inibidores , Carga Tumoral/efeitos dos fármacos , Administração Oral , Animais , Morte Celular/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Microambiente Celular/efeitos dos fármacos , Quimiocinas/administração & dosagem , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Humanos , Imunocompetência/efeitos dos fármacos , Inflamação/tratamento farmacológico , Inflamação/patologia , Camundongos , Camundongos Endogâmicos C57BL , Modelos Biológicos , Monócitos/efeitos dos fármacos , Monócitos/metabolismo , Mieloma Múltiplo/complicações , Mieloma Múltiplo/patologia , Osteoclastos/efeitos dos fármacos , Osteoclastos/metabolismo , Osteoclastos/patologia , Osteólise/complicações , Osteólise/patologia , Ratos , Receptores CCR1/metabolismo
13.
Peptides ; 32(9): 1866-71, 2011 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-21855588

RESUMO

Visceral adipose tissue-derived serpin (vaspin) improves glucose tolerance and insulin sensitivity in diet-induced obese mice. Chemerin may increase insulin sensitivity in adipose tissue and seems to be associated with several key aspects of metabolic syndrome. Decreased levels of omentin-1 are associated with increasing obesity and insulin resistance. Our study aimed to investigate the effects of vaspin, chemerin and omentin-1 acute administration on feeding and hypothalamic gene expression of peptides which play a key role in feeding regulation. 35 rats were injected into the arcuate nucleus (ARC) of the hypothalamus with either saline (n=8), vaspin (1µg/kg; n=9), chemerin (8µg/kg; n=9), or omentin-1 (8µg/kg; n=9). Food intake in the following 24h was recorded, thereafter rats were sacrificed. Total RNA was extracted from hypothalami and reverse transcribed to evaluate hypothalamic gene expression of agouti-related peptide (AgRP), neuropeptide Y (NPY), orexin-A, cocaine- and amphetamine-regulated transcript (CART), corticotrophin releasing hormone (CRH) and proopiomelanocortin (POMC), by real-time reverse transcription polymerase chain reaction. Compared to vehicle, vaspin injection significantly decreased feeding, while chemerin and omentin-1 had no effect in the tested dose. Vaspin treatment significantly decreased NPY and increased POMC gene expression. Chemerin treatment led to a significant increase of both AgRP and POMC gene expression. Omentin-1 treatment did not modify gene expression of the investigated peptides. Therefore, vaspin is an adipokine triggering anorectic pathways in the hypothalamus, where reduction of NPY and increase of POMC mRNA levels mediate feeding inhibition. Chemerin and omentin-1 have no effect on feeding in the tested dose.


Assuntos
Quimiocinas/farmacologia , Citocinas/farmacologia , Comportamento Alimentar/efeitos dos fármacos , Hipotálamo/efeitos dos fármacos , Lectinas/farmacologia , Serpinas/farmacologia , Proteína Relacionada com Agouti/genética , Proteína Relacionada com Agouti/metabolismo , Animais , Depressores do Apetite/farmacologia , Quimiocinas/administração & dosagem , Citocinas/administração & dosagem , Ingestão de Alimentos/efeitos dos fármacos , Proteínas Ligadas por GPI/administração & dosagem , Proteínas Ligadas por GPI/farmacologia , Regulação da Expressão Gênica , Humanos , Hipotálamo/citologia , Peptídeos e Proteínas de Sinalização Intercelular , Lectinas/administração & dosagem , Masculino , Camundongos , Neuropeptídeo Y/genética , Neuropeptídeo Y/metabolismo , Neuropeptídeos/genética , Neuropeptídeos/metabolismo , Reação em Cadeia da Polimerase/métodos , Pró-Opiomelanocortina/genética , Pró-Opiomelanocortina/metabolismo , Ratos , Ratos Wistar , Proteínas Recombinantes/administração & dosagem , Proteínas Recombinantes/farmacologia , Serpinas/administração & dosagem
14.
J Control Release ; 155(2): 184-92, 2011 Oct 30.
Artigo em Inglês | MEDLINE | ID: mdl-21708196

RESUMO

Patients with malignant non-Hodgkin's lymphomas (NHL) of B-cell lineages relapse despite initial anti-tumor response to chemotherapy or antibody treatments. Failure to eliminate the tumor is often because of inadequate priming, low cell numbers and suboptimal phenotype of effector T cells. Here we describe a new biomaterial-based controlled-release paradigm to treat weakly immunogenic NHLs by in-situ amplifying the number of functional, antigen-specific T-helper 1 (Th1) cells following immunotherapy. An injectable, synthetic immune priming center (sIPC) consisting of an in-situ crosslinking, chemokine-carrying hydrogel and both DNA- and siRNA dual-loaded microparticles, is reported. This sIPC chemo attracts a large number of immature dendritic cells (DCs) at the site of administration and efficiently co-delivers both DNA antigens and interleukin-10 (IL10)-silencing siRNA to those cells. Using a murine model of A20 B cell lymphoma, we demonstrate that combination of DNA-antigen delivery and IL10 silencing, synergistically activate recruited immature DCs and cause a strong shift towards Th1 response while suppressing Th2 and Th17 cytokines. sIPC-based immunotherapy showed 45% more CD8+ cytotoxic T cell (CTL) response and 53% stronger CD4+ CTL activity compared to naked DNA vaccine. In addition, in-vivo sIPC immunization induced significant protection (p<0.01) against subsequent tumor challenge. Such a multi-modal, injectable system that simultaneously delivers chemokines, siRNA and DNA antigens to DCs marks a new approach to in-situ priming and modulation during immunotherapy and could provide effective vaccination strategies against cancers and infectious diseases.


Assuntos
Vacinas Anticâncer/administração & dosagem , Quimiocinas/administração & dosagem , DNA/administração & dosagem , Portadores de Fármacos/química , Linfoma de Células B/imunologia , RNA Interferente Pequeno/administração & dosagem , Células Th1 , Animais , Antígenos de Neoplasias/imunologia , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD8-Positivos/imunologia , Vacinas Anticâncer/imunologia , Quimiocinas/imunologia , Reagentes para Ligações Cruzadas , DNA/imunologia , Preparações de Ação Retardada , Células Dendríticas/citologia , Células Dendríticas/imunologia , Feminino , Hidrogéis , Idiotipos de Imunoglobulinas/imunologia , Injeções Intramusculares , Interleucina-10/imunologia , Linfoma de Células B/terapia , Camundongos , Camundongos Endogâmicos BALB C , RNA Interferente Pequeno/imunologia , Linfócitos T Citotóxicos/imunologia , Células Th1/citologia , Células Th1/imunologia
15.
J Immunol ; 186(4): 2219-28, 2011 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-21242526

RESUMO

Exosomes derived from dendritic cells or tumor cells are a population of nanometer-sized membrane vesicles that can induce specific antitumor immunity. During investigation of the effects of hyperthermia on antitumor immune response, we found that exosomes derived from heat-stressed tumor cells (HS-TEX) could chemoattract and activate dendritic cells (DC) and T cells more potently than that by conventional tumor-derived exosomes. We show that HS-TEX contain chemokines, such as CCL2, CCL3, CCL4, CCL5, and CCL20, and the chemokine-containing HS-TEX are functionally competent in chemoattracting CD11c(+) DC and CD4(+)/CD8(+) T cells both in vitro and in vivo. Moreover, the production of chemokine-containing HS-TEX could be inhibited by ATP inhibitor, calcium chelator, and cholesterol scavenger, indicating that the mobilization of chemokines into exosomes was ATP- and calcium-dependent and via a lipid raft-dependent pathway. We consistently found that the intracellular chemokines could be enriched in lipid rafts after heat stress. Accordingly, intratumoral injection of HS-TEX could induce specific antitumor immune response more efficiently than that by tumor-derived exosomes, thus inhibiting tumor growth and prolonging survival of tumor-bearing mice more significantly. Therefore, our results demonstrate that exosomes derived from HS-TEX represent a kind of efficient tumor vaccine and can chemoattract and activate DC and T cells, inducing more potent antitumor immune response. Release of chemokines through exosomes via lipid raft-dependent pathway may be a new method of chemokine exocytosis.


Assuntos
Vacinas Anticâncer/imunologia , Carcinoma Pulmonar de Lewis/prevenção & controle , Quimiocinas/metabolismo , Exossomos/metabolismo , Transtornos de Estresse por Calor/prevenção & controle , Melanoma Experimental/prevenção & controle , Microdomínios da Membrana/imunologia , Transdução de Sinais/imunologia , Animais , Vacinas Anticâncer/administração & dosagem , Vacinas Anticâncer/metabolismo , Carcinoma Pulmonar de Lewis/imunologia , Carcinoma Pulmonar de Lewis/metabolismo , Linhagem Celular Tumoral , Células Cultivadas , Quimiocinas/administração & dosagem , Quimiocinas/imunologia , Técnicas de Cocultura , Células Dendríticas/imunologia , Células Dendríticas/metabolismo , Exossomos/imunologia , Transtornos de Estresse por Calor/imunologia , Transtornos de Estresse por Calor/metabolismo , Temperatura Alta , Masculino , Melanoma Experimental/imunologia , Melanoma Experimental/metabolismo , Microdomínios da Membrana/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Subpopulações de Linfócitos T/imunologia , Subpopulações de Linfócitos T/metabolismo
16.
Proc Natl Acad Sci U S A ; 108(1): 67-72, 2011 Jan 04.
Artigo em Inglês | MEDLINE | ID: mdl-21149682

RESUMO

Porous biomaterials have been widely used as scaffolds in tissue engineering and cell-based therapies. The release of biological agents from conventional porous scaffolds is typically governed by molecular diffusion, material degradation, and cell migration, which do not allow for dynamic external regulation. We present a new active porous scaffold that can be remotely controlled by a magnetic field to deliver various biological agents on demand. The active porous scaffold, in the form of a macroporous ferrogel, gives a large deformation and volume change of over 70% under a moderate magnetic field. The deformation and volume variation allows a new mechanism to trigger and enhance the release of various drugs including mitoxantrone, plasmid DNA, and a chemokine from the scaffold. The porous scaffold can also act as a depot of various cells, whose release can be controlled by external magnetic fields.


Assuntos
Materiais Biocompatíveis , Sistemas de Liberação de Medicamentos/métodos , Géis/uso terapêutico , Magnetismo/métodos , Polímeros/uso terapêutico , Tecidos Suporte , Quimiocinas/administração & dosagem , DNA/administração & dosagem , Preparações de Ação Retardada/uso terapêutico , Mitoxantrona/administração & dosagem , Porosidade
17.
Cancer J ; 16(4): 325-35, 2010.
Artigo em Inglês | MEDLINE | ID: mdl-20693843

RESUMO

Chemokines (ie, chemoattractant cytokines) are a family of small secreted molecules that mediate leukocyte migration. It is becoming increasingly more evident that chemokines play an integral role in the initiation of a specific immune response. With respect to cancer, chemokines are being studied for both their role in tumor biology and as promising immunotherapy candidates. We review several areas of chemokine importance in tumor immunity and discuss the experimental evidence that is leading to the clinical use of this cytokine family in new treatment approaches for patients with cancer.


Assuntos
Quimiocinas/imunologia , Neoplasias/imunologia , Neoplasias/terapia , Animais , Quimiocinas/administração & dosagem , Quimiocinas/genética , Quimiocinas/metabolismo , Humanos , Receptores de Quimiocinas/imunologia
19.
Brain Res ; 1308: 24-34, 2010 Jan 13.
Artigo em Inglês | MEDLINE | ID: mdl-19857473

RESUMO

Chemokine-like factor 1 (CKLF1) is a newly cloned chemotactic cytokine. The roles of CKLF1 in the immune system and the respiratory system have been reported, but its function in the nervous system is still remaining unclear. We aimed to investigate the role of CKLF1 in the nerve cell migration and its regulatory mechanisms. By chemotaxis assays and wound-healing assays, CKLF1 stimulated the migration of SH-SY5Y cells dose-dependently. By immunofluorescence staining, CKLF1 induced actin polymerization. By western blotting, proline-rich tyrosine kinase 2 (PYK2) was phosphorylated at Tyr-402 in response to CKLF1 and this phosphorylation was apparently suppressed by phospholipase C-gamma inhibitor U73122, but not extracellular Ca(2+) chelator EGTA. Furthermore, after transfection of dominant-negative mutant PYK2 plasmid, the chemotaxis upon CKLF1 was significantly attenuated in SH-SY5Y cells. Concluding, CKLF1 stimulates the migration of SH-SY5Y cells dose-dependently by activating non-extracellular Ca(2+)-dependent tyrosine kinases pathway and inducing actin polymerization.


Assuntos
Cálcio/metabolismo , Movimento Celular/efeitos dos fármacos , Quimiocinas/administração & dosagem , Quinase 2 de Adesão Focal/metabolismo , Neurônios/efeitos dos fármacos , Actinas/metabolismo , Análise de Variância , Western Blotting , Adesão Celular/efeitos dos fármacos , Linhagem Celular , Células Cultivadas , Citoesqueleto/efeitos dos fármacos , Citoesqueleto/metabolismo , Relação Dose-Resposta a Droga , Inibidores Enzimáticos/farmacologia , Estrenos/farmacologia , Imunofluorescência , Quinase 2 de Adesão Focal/genética , Humanos , Proteínas com Domínio MARVEL , Fosfolipase C gama/metabolismo , Fosforilação/efeitos dos fármacos , Pirrolidinonas/farmacologia , Transdução de Sinais/efeitos dos fármacos , Transfecção , Cicatrização/efeitos dos fármacos
20.
Biomaterials ; 30(28): 5187-200, 2009 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-19560815

RESUMO

Polymer-based, injectable systems that can simultaneously deliver multiple bioactive agents in a controlled manner could significantly enhance the efficacy of next generation therapeutics. For immunotherapies to be effective, both prophylactically or therapeutically, it is not only critical to drive the antigen (Ag)-specific immune response strongly towards either T helper type 1 (Th1) or Th2 phenotype, but also to promote recruitment of a high number of antigen-presenting cells (APCs) at the site of immunization. We have recently reported a microparticle-based system capable of simultaneously delivering siRNA and DNA to APCs. Here we present an in-situ crosslinkable, injectable formulation containing dendritic cell (DC)-chemo-attractants and dual-mode DNA-siRNA loaded microparticles to attract immature DCs and simultaneously deliver, to the migrated cells, immunomodulatory siRNA and plasmid DNA antigens. These low crosslink density hydrogels were designed to degrade within 2-7 days in vitro and released chemokines in a sustained manner. Chemokine carrying gels attracted 4-6 folds more DCs over a sustained period in vitro, compared to an equivalent bolus dose. Interestingly, migrated DCs were able to infiltrate the hydrogels and efficiently phagocytose the siRNA-DNA carrying microparticles. Hydrogel embedded microparticles co-delivering Interleukin-10 siRNA and plasmid DNA antigens exhibited efficient Interleukin-10 gene knockdown in migrated primary DCs in vitro.


Assuntos
Quimiocinas/administração & dosagem , DNA/administração & dosagem , Células Dendríticas/citologia , Portadores de Fármacos/química , Hidrogéis/química , RNA Interferente Pequeno/administração & dosagem , Animais , Células da Medula Óssea/citologia , Células da Medula Óssea/imunologia , Células Cultivadas , Quimiocinas/imunologia , Reagentes para Ligações Cruzadas , DNA/imunologia , Células Dendríticas/imunologia , Feminino , Técnicas de Silenciamento de Genes , Interleucina-10/genética , Interleucina-10/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Plasmídeos/administração & dosagem , Plasmídeos/imunologia , RNA Interferente Pequeno/imunologia
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