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1.
Resusc Plus ; 17: 100538, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-38205148

RESUMO

Background: International guidelines for neuroprotection following out-of-hospital cardiac arrest (OHCA) recommend fever prevention ahead of routine temperature management. This study aimed to identify any effect of changing from targeted temperature management to fever prevention on neurological outcome following OHCA. Methods: A retrospective observational cohort study was conducted of consecutive admissions to an ICU at a tertiary OHCA centre. Comparison was made between a period of protocolised targeted temperature management (TTM) to 36 °C and a period of fever prevention. Results: Data were available for 183 patients. Active temperature management was administered in 86/118 (72%) of the TTM cohort and 20/65 (31%) of the fever prevention group. The median highest temperature prior to the start of temperature management was significantly lower in the TTM group at 35.6 (IQR 34.9-36.2) compared to 37.9 °C (IQR 37.7-38.2) in the fever prevention group (adjusted p < 0.001).There was no difference in the proportion of patients discharged with Cerebral Performance Category 1 or 2 between the groups (42% vs. 40%, p = 0.88). Patients in the fever prevention group required a reduced duration of noradrenaline (36 vs. 46 h, p = 0.03) and a trend towards a reduced duration of propofol (37 vs. 56 h, p = 0.06).In unadjusted analysis, use of active temperature management (irrespective of group) appeared to be associated with decreased risk of poor outcome (OR = 0.43, 95% CI 0.23-0.78) but after adjustment for patient age, presenting rhythm, witnessed arrest and duration of CPR, this was no longer significant (OR = 0.93, 95% CI 0.37-2.31, p = 0.88). Conclusion: Switching from TTM to fever prevention following OHCA was associated with similar rates of neurological outcomes, with a possible decrease in sedation and vasopressor requirements.

2.
J Neuroimmunol ; 209(1-2): 33-9, 2009 Apr 30.
Artigo em Inglês | MEDLINE | ID: mdl-19232440

RESUMO

Based on gene expression data, we tested the P8A-CCL2 variant of the chemokine CCL2, able to interfere with the chemotactic properties of the parental molecule, in relapsing-remitting (RR)-EAE SJL. Only preventive treatment significantly delayed disease onset in a dose dependent manner. P8A-CCL2 administration, however, decreased demyelination, axonal loss and number of CNS infiltrating T cells and macrophages. Immunological analysis revealed that P8A-CCL2 does not act on Ag-specific T cell proliferation and does not interfere with the differentiation of IFNgamma-releasing effectors T cells. These results suggest that the therapeutic mechanism of P8A-CCL2 may rely on interference with immune cell recruitment.


Assuntos
Quimiocina CCL2/farmacologia , Quimiotaxia de Leucócito/efeitos dos fármacos , Encefalomielite Autoimune Experimental/tratamento farmacológico , Encefalomielite Autoimune Experimental/imunologia , Bainha de Mielina/efeitos dos fármacos , Adulto , Animais , Proliferação de Células/efeitos dos fármacos , Quimiocina CCL2/síntese química , Quimiocina CCL2/uso terapêutico , Quimiotaxia de Leucócito/imunologia , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Encefalomielite Autoimune Experimental/fisiopatologia , Feminino , Humanos , Fatores Imunológicos/farmacologia , Fatores Imunológicos/uso terapêutico , Interferon gama/metabolismo , Macrófagos/efeitos dos fármacos , Macrófagos/imunologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Pessoa de Meia-Idade , Bainha de Mielina/imunologia , Bainha de Mielina/patologia , Fármacos Neuroprotetores/farmacologia , Fármacos Neuroprotetores/uso terapêutico , Linfócitos T/efeitos dos fármacos , Linfócitos T/imunologia , Degeneração Walleriana/tratamento farmacológico , Degeneração Walleriana/imunologia , Degeneração Walleriana/fisiopatologia
3.
J R Coll Physicians Edinb ; 39(4): 357-61, 2009 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-20509461

RESUMO

Henry Matthew was appointed a consultant in the Royal Infirmary of Edinburgh in 1955, by which time he was a highly regarded general physician with an interest in cardiology. In 1964 he agreed, almost certainly reluctantly, to head the recently designated Regional Poisoning Treatment Centre, which he did until his retirement ten years later. Matthew quickly established himself as an authority in clinical toxicology, mainly from an unrivalled experience of treating poisoned patients, day-in and day-out, but also by publishing original research, letters and books. Such were his contributions that he is regarded as the father of clinical toxicology.


Assuntos
Toxicologia/história , História do Século XX , História do Século XXI , Escócia
4.
Biochem Soc Trans ; 34(Pt 3): 422-6, 2006 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-16709177

RESUMO

Chemokines exert their biological activity through high-affinity interactions with cell-surface receptors, thereby activating specific signalling pathways, and a second low-affinity interaction with proteoglycans. Proteoglycans consist of a protein core, to which GAG (glycosaminoglycan) chains are attached. The GAGs are long, linear, sulphated and highly charged heterogeneous polysaccharides that are expressed throughout the body in different forms depending on the developmental or pathological state of the organ/organism. Mechanistically, the GAG interaction is thought to facilitate the retention of chemokines on cell surfaces, thereby forming a high local concentration required for cell activation. Recently, we demonstrated that certain chemokines require interactions with GAGs for their in vivo function. Additionally we have shown that chemokines oligomerize on immobilized GAGs, and this ability to form higher order oligomers has also been shown to be essential for the activity of certain chemokines in vivo. We believe that interference with the chemokine-GAG interaction provides a novel anti-inflammatory strategy, exemplified by a variant of RANTES (regulated upon activation, normal T-cell expressed and secreted) that has abrogated GAG binding and oligomerization properties.


Assuntos
Quimiocinas/metabolismo , Proteoglicanas/metabolismo , Animais , Quimiocinas/fisiologia , Humanos , Proteoglicanas/fisiologia
5.
Cytokine Growth Factor Rev ; 16(6): 625-36, 2005 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-15990353

RESUMO

Despite their key role in inflammation, the apparent redundancy in the chemokine system is often cited as an argument against probing chemokines as therapeutic targets for inflammation. However, this in vitro redundancy frequently does not translate to the in vivo situation, as exemplified by the use of specific receptor antagonists, ligand neutralizing or receptor blocking antibodies and gene-deleted mice in models of human disease. Specificity may be conferred onto the chemokine system by fine-tuning of responses both temporally and spatially through their highly specific interactions with glycosaminoglycans (GAGs). In this survey, we present evidence for specificity in the interaction and introduce emerging technologies that enable detailed assessment of protein-GAG interactions. Finally, we address the issue of exploitation of this interaction for therapeutic advantage.


Assuntos
Quimiocinas/metabolismo , Sistemas de Liberação de Medicamentos , Glicosaminoglicanos/metabolismo , Doenças do Sistema Imunitário/tratamento farmacológico , Glicosaminoglicanos/química , Glicosaminoglicanos/classificação , Humanos , Modelos Moleculares
6.
Annu Rev Biochem ; 74: 385-410, 2005.
Artigo em Inglês | MEDLINE | ID: mdl-15952892

RESUMO

Immune modulators such as cytokines and growth factors exert their biological activity through high-affinity interactions with cell-surface receptors, thereby activating specific signaling pathways. However, many of these molecules also participate in low-affinity interactions with another class of molecules, referred to as proteoglycans. Proteoglycans consist of a protein core to which glycosaminoglycan (GAG) chains are attached. The GAGs are long, linear, sulfated, and highly charged heterogeneous polysaccharides that are expressed throughout the body in different forms, depending on the developmental or pathological state of the organ/organism. They participate in many biological functions, including organogenesis and growth control, cell adhesion, signaling, inflammation, tumorigenesis, and interactions with pathogens. Recently, it was demonstrated that certain chemokines require interactions with GAGs for their in vivo function. The GAG interaction is thought to provide a mechanism for retaining chemokines on cell surfaces, facilitating the formation of chemokine gradients. These gradients serve as directional cues to guide the migration of the appropriate cells in the context of their inflammatory, developmental, and homeostatic functions. In this review, we discuss GAGs and their interaction with proteins, with a special emphasis on the chemokine system.


Assuntos
Quimiocinas/metabolismo , Glicosaminoglicanos/metabolismo , Proteínas/metabolismo , Animais , Sequência de Carboidratos , Quimiocinas/química , Quimiocinas/genética , Glicosaminoglicanos/química , Glicosaminoglicanos/genética , Humanos , Técnicas In Vitro , Substâncias Macromoleculares , Modelos Moleculares , Dados de Sequência Molecular , Estrutura Molecular , Proteínas/química , Proteínas/genética , Proteoglicanas/química , Proteoglicanas/genética , Proteoglicanas/metabolismo
7.
Clin Med (Lond) ; 5(6): 621-5, 2005.
Artigo em Inglês | MEDLINE | ID: mdl-16411359

RESUMO

Levels of staffing and access to diagnostics at weekends are recognised to be significantly lower than on weekdays. It is unclear if subsequent inpatient mortality and readmission rates for acute medical admissions are increased for weekend admissions compared to those on a weekday. A large Canadian study demonstrated increased weekend mortality but does the Edinburgh healthcare model support these findings? This study analysed all hospital admissions in 2001 to the Royal Infirmary of Edinburgh for six predetermined diagnoses (total 3,244): chronic obstructive pulmonary disease, cerebrovascular accidents, pulmonary embolism, pneumonia, collapse and upper gastrointestinal bleed. We compared hospital mortality rates, readmission rates and hospital length of stay for weekend admissions as compared to those on a weekday. Weekend admission was not associated with significantly higher in-hospital mortality, readmission rates or increased length of stay compared to the weekday equivalent for any of the six conditions. The implementation of an acute medical admissions unit in the Royal Infirmary of Edinburgh, with consistent staffing levels and 24-hour access to diagnostics for the early phase of critical illness, may have helped address the discrepancy in care suggested by previous studies.


Assuntos
Emergências/classificação , Serviço Hospitalar de Emergência/estatística & dados numéricos , Acessibilidade aos Serviços de Saúde , Mortalidade Hospitalar , Admissão do Paciente/estatística & dados numéricos , Admissão e Escalonamento de Pessoal , Idoso , Idoso de 80 Anos ou mais , Serviços de Diagnóstico/estatística & dados numéricos , Serviços de Diagnóstico/provisão & distribuição , Emergências/epidemiologia , Feminino , Férias e Feriados , Humanos , Masculino , Pessoa de Meia-Idade , Periodicidade , Escócia/epidemiologia , Fatores de Tempo , Resultado do Tratamento , Recursos Humanos
8.
J Toxicol Clin Toxicol ; 42(1): 1-26, 2004.
Artigo em Inglês | MEDLINE | ID: mdl-15083932

RESUMO

This Position Paper was prepared using the methodology agreed by the American Academy of Clinical Toxicology (AACT) and the European Association of Poisons Centres and Clinical Toxicologists (EAPCCT). All relevant scientific literature was identified and reviewed critically by acknowledged experts using set criteria. Well-conducted clinical and experimental studies were given precedence over anecdotal case reports and abstracts were not considered. A draft Position Paper was then produced and presented at the North American Congress of Clinical Toxicology in October 2001 and at the EAPCCT Congress in May 2002 to allow participants to comment on the draft after which a revised draft was produced. The Position Paper was subjected to detailed peer review by an international group of clinical toxicologists chosen by the AACT and the EAPCCT, and a final draft was approved by the boards of the two societies. The Position Paper includes a summary statement (Position Statement) for ease of use, which will also be published separately, as well as the detailed scientific evidence on which the conclusions of the Position Paper are based. Urine alkalinization is a treatment regimen that increases poison elimination by the administration of intravenous sodium bicarbonate to produce urine with a pH > or = 7.5. The term urine alkalinization emphasizes that urine pH manipulation rather than a diuresis is the prime objective of treatment; the terms forced alkaline diuresis and alkaline diuresis should therefore be discontinued. Urine alkalinization increases the urine elimination of chlorpropamide, 2,4-dichlorophenoxyacetic acid, diflunisal, fluoride, mecoprop, methotrexate, phenobarbital, and salicylate. Based on volunteer and clinical studies, urine alkalinization should be considered as first line treatment for patients with moderately severe salicylate poisoning who do not meet the criteria for hemodialysis. Urine alkalinization cannot be recommended as first line treatment in cases of phenobarbital poisoning as multiple-dose activated charcoal is superior. Supportive care, including the infusion of dextrose, is invariably adequate in chlorpropamide poisoning. A substantial diuresis is required in addition to urine alkalinization in the chlorophenoxy herbicides, 2,4-dichlorophenoxyacetic acid, and mecoprop, if clinically important herbicide elimination is to be achieved. Volunteer studies strongly suggest that urine alkalinization increases fluoride elimination, but this is yet to be confirmed in clinical studies. Although urine alkalinization is employed clinically in methotrexate toxicity, currently there is only one study that supports its use. Urine alkalinization enhances diflunisal excretion, but this technique is unlikely to be of value in diflunisal poisoning. In conclusion, urine alkalinization should be considered first line treatment in patients with moderately severe salicylate poisoning who do not meet the criteria for hemodialysis. Urine alkalinization and high urine flow (approximately 600 mL/h) should also be considered in patients with severe 2,4-dichlorophenoxyacetic acid and mecoprop poisoning. Administration of bicarbonate to alkalinize the urine results in alkalemia (an increase in blood pH or reduction in its hydrogen ion concentration); pH values approaching 7.70 have been recorded. Hypokalemia is the most common complication but can be corrected by giving potassium supplements. Alkalotic tetany occurs occasionally, but hypocalcemia is rare. There is no evidence to suggest that relatively short-duration alkalemia (more than a few hours) poses a risk to life in normal individuals or in those with coronary and cerebral arterial disease.


Assuntos
Álcalis/urina , Centros de Controle de Intoxicações , Guias de Prática Clínica como Assunto , Toxicologia/métodos , Animais , Europa (Continente) , Humanos , Concentração de Íons de Hidrogênio , Sociedades Médicas , Estados Unidos
9.
Biochem Soc Trans ; 32(Pt 2): 366-77, 2004 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-15046611

RESUMO

Chemokines are small chemoattractant cytokines that control a wide variety of biological and pathological processes, ranging from immunosurveillance to inflammation, and from viral infection to cancer. Genetic and pharmacological studies have shown that chemokines are responsible for the excessive recruitment of leucocytes to inflammatory sites and damaged tissue. In the present paper, we discuss the rationale behind interfering with the chemokine system and introduce various points for therapeutic intervention using either protein-based or small-molecule inhibitors. Unlike other cytokines, chemokines signal via seven-transmembrane GPCRs (G-protein-coupled receptors), which are favoured targets by the pharmaceutical industry, and, as such, they are the first cytokines for which small-molecule-receptor antagonists have been developed. In addition to the high-affinity receptor interaction, chemokines have an in vivo requirement to bind to GAGs (glycosaminoglycans) in order to mediate directional cell migration. Prevention of the GAG interaction has been shown to be a viable therapeutic strategy. Targeting chemokine intracellular signalling pathways offers an alternative small-molecule approach. One of the key signalling targets downstream of a variety of chemokine receptors identified to date is PI3Kgamma (phosphoinositide 3-kinase gamma), a member of the class I PI3K family. Thus the chemokine system offers many potential entry points for innovative anti-inflammatory therapies for autoimmune diseases, such as multiple sclerosis, rheumatoid arthritis and allergic contact dermatitis.


Assuntos
Quimiocinas/antagonistas & inibidores , Animais , Bioquímica/métodos , Glicosaminoglicanos/metabolismo , Humanos , Ligantes , Camundongos , Camundongos Knockout , Modelos Biológicos , Modelos Químicos , Fosfatidilinositol 3-Quinases/metabolismo , Receptores Acoplados a Proteínas G/metabolismo , Transdução de Sinais
10.
Br J Surg ; 90(6): 698-704, 2003 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-12808617

RESUMO

BACKGROUND: Severe acute pancreatitis leads to a systemic inflammatory response characterized by widespread leucocyte activation and, as a consequence, distant lung injury. In CC chemokines the first two cysteine residues are adjacent to each other. The aim of this study was to evaluate the effect of Met-RANTES, a CC chemokine receptor antagonist, on pancreatic inflammation and lung injury in caerulein-induced acute pancreatitis in mice. METHODS: Acute pancreatitis was induced in mice by hourly intraperitoneal injection of caerulein. Met-RANTES was administered either 30 min before or 1 h after starting caerulein injections, and pancreatic inflammation and lung injury were assessed. There were five groups of eight mice each including controls. RESULTS: Treatment with Met-RANTES had little effect on caerulein-induced pancreatic damage. Met-RANTES, however, reduced lung injury when given either before administration of caerulein (mean(s.e.m.) lung myeloperoxidase (MPO) 1.47(0.19) versus 3.70(0.86)-fold increase over control, P = 0.024; mean(s.e.m.) microvascular permeability 1.15(0.05) versus 3.57(0.63) lavage to plasma fluorescein isothiocyanate-labelled albumin fluorescence ratio (L/P) per cent, P = 0.002) or after caerulein administration (lung MPO 1.96(0.27) versus 3.65(0.63)-fold increase over control, P = 0.029; microvascular permeability 0.94(0.04) versus 2.85(0.34) L/P per cent, P < 0.001). CONCLUSION: Treatment with Met-RANTES reduces lung damage associated with caerulein-induced pancreatitis in mice. Chemokine receptor antagonists may be of use for the treatment of the systemic complications of acute pancreatitis.


Assuntos
Anti-Inflamatórios/administração & dosagem , Quimiocina CCL5/análogos & derivados , Quimiocina CCL5/administração & dosagem , Pneumopatias/prevenção & controle , Pancreatite/complicações , Síndrome de Resposta Inflamatória Sistêmica/prevenção & controle , Doença Aguda , Animais , Ceruletídeo/toxicidade , Avaliação Pré-Clínica de Medicamentos , Pneumopatias/patologia , Camundongos , Microcirculação , Pancreatite/patologia , Receptores de Quimiocinas/antagonistas & inibidores
12.
Blood ; 98(12): 3476-8, 2001 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-11719391

RESUMO

Macrophage inflammatory protein-1alpha (MIP-1alpha) is a member of the chemokine family of proinflammatory mediators. In addition to its inflammatory roles, MIP-1alpha has been shown to be active as an inhibitor of primitive hemopoietic cell proliferation. Indeed, a dysfunction in this inhibitory process has been postulated to contribute to leukemogenesis. Research has been aimed at characterizing the receptor involved in cellular inhibition by MIP-1alpha. This study demonstrates that of all the beta-chemokines tested, only MIP-1alpha is capable of inhibiting primitive hemopoietic cell proliferation. Because no MIP-1alpha-specific receptors have been identified, this suggests that inhibition is mediated by an uncharacterized receptor. Further evidence for the involvement of a novel receptor in this process is the equivalent potencies of MIP-1alphaS and MIP-1alphaP variants of human MIP-1alpha and the fact that primitive cells from bone marrow derived from individual MIP-1alpha receptor null mice display a full response to MIP-1alpha inhibition.


Assuntos
Divisão Celular/efeitos dos fármacos , Células-Tronco Hematopoéticas/citologia , Proteínas Inflamatórias de Macrófagos/farmacologia , Receptores de Quimiocinas/fisiologia , Animais , Células da Medula Óssea/citologia , Quimiocina CCL3 , Quimiocina CCL4 , Camundongos , Camundongos Knockout , Receptores de Quimiocinas/deficiência
13.
J Exp Med ; 194(10): 1541-7, 2001 Nov 19.
Artigo em Inglês | MEDLINE | ID: mdl-11714760

RESUMO

The chemokine thymus and activation-regulated chemokine (TARC; CCL17) is displayed by cutaneous (but not intestinal) venules, and is thought to trigger vascular arrest of circulating skin homing memory T cells, which uniformly express the TARC receptor CC chemokine receptor (CCR)4. Cutaneous T cell-attracting chemokine (CTACK; CCL27), expressed by skin keratinocytes, also attracts cutaneous memory T cells, and is hypothesized to assist in lymphocyte recruitment to skin as well. Here we show that chronic cutaneous inflammation induces CD4 T cells expressing E-selectin binding activity (a marker of skin homing memory cells) in draining lymph node, and that these E-selectin ligand+ T cells migrate efficiently to TARC and to CTACK. In 24 h in vivo homing assays, stimulated lymph node T cells from wild-type mice or, surprisingly, from CCR4-deficient donors migrate efficiently to inflamed skin; and an inhibitory anti-CTACK antibody has no effect on wild-type lymphocyte recruitment. However, inhibition with anti-CTACK monoclonal antibody abrogates skin recruitment of CCR4-deficient T cells. We conclude that CTACK and CCR4 can both support homing of T cells to skin, and that either one or the other is required for lymphocyte recruitment in cutaneous delayed type hypersensitivity.


Assuntos
Quimiocinas CC/fisiologia , Dermatite/imunologia , Linfócitos/fisiologia , Receptores de Quimiocinas/metabolismo , Receptores de Quimiocinas/fisiologia , Transferência Adotiva , Animais , Movimento Celular , Quimiocina CCL17 , Feminino , Ligantes , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Ratos , Receptores CCR10 , Receptores CCR4
14.
Curr Opin Pharmacol ; 1(4): 417-24, 2001 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-11710742

RESUMO

Chemokines are cytokines that specifically direct the trafficking of immune cells in the body. They offer a novel point of therapeutic intervention, as inhibiting specific chemokines and receptors could prevent the excessive recruitment of leukocytes to sites of inflammation. This approach could be considered to act upstream of the therapies used today which, for the most part, act on the cells already at the site of inflammation. The receptors for chemokines are G-protein-coupled seven-transmembrane receptors, which are particularly tractable for the pharmaceutical industry. The search for small-molecule inhibitors of these receptors has been fruitful and the numbers of patents and, more recently, peer-reviewed publications are growing rapidly. The first clinical trial was initiated this year, so although it is too soon to be able to report these results we hope to see the outcome of this research in the near future.


Assuntos
Quimiocinas/fisiologia , Sistema Imunitário/fisiologia , Animais , Asma/tratamento farmacológico , Rejeição de Enxerto/imunologia , HIV/efeitos dos fármacos , Humanos , Esclerose Múltipla/tratamento farmacológico
15.
J Immunol ; 167(6): 3443-53, 2001 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-11544337

RESUMO

Cytokines and chemokines are responsible for the attraction and activation of eosinophils in allergic and inflammatory diseases. Whereas cytokines such as IL-3, IL-5, and GM-CSF activate eosinophils via heterodimeric receptors containing a distinct alpha-chain (binding domain) and a common beta-chain (signaling domain), chemokines such as eotaxin activate eosinophils via seven-transmembrane G(i) protein-coupled CCRs. Recent studies have demonstrated the importance of CCR3 on human eosinophils that undergo receptor recycling after chemokine activation, but the modulation of this receptor by cytokines has not yet been addressed. In this study, we demonstrate that IL-3 induces a dose- and time-dependent down-regulation of CCR3 from the surface of human eosinophils comparable to the CCR3-specific ligand eotaxin, whereas IL-5, GM-CSF, IL-4, IL-10, IL-13, IFN-gamma, and TNF-alpha had no effect. Maximal down-regulation of CCR3 in response to IL-3 was reached at 24 h. Reduction of CCR3 surface protein in response to IL-3 could be prevented by an anti-IL-3 mAb and was neither due to the release of CC chemokines nor to nonspecific binding of IL-3 to CCR3. Moreover, down-regulation was prevented by phenylarsine oxide, a nonspecific inhibitor of receptor internalization. After 24 h, IL-3-induced decrease of CCR3 surface expression correlated with diminished mRNA expression, suggesting a transcriptional regulation mechanism. Since wortmannin partially inhibited IL-3- but not eotaxin-induced CCR3 down-regulation, receptor down-modulation seems to underlie different signaling events. Therefore, these data suggest a novel role for the cytokine IL-3 in the activation process of eosinophils and its predominant chemokine receptor CCR3.


Assuntos
Quimiocinas CC , Regulação para Baixo/efeitos dos fármacos , Eosinófilos/efeitos dos fármacos , Interleucina-3/farmacologia , RNA Mensageiro/biossíntese , Receptores de Quimiocinas/biossíntese , Androstadienos/farmacologia , Animais , Arsenicais/farmacologia , Sinalização do Cálcio/efeitos dos fármacos , Células Cultivadas , Quimiocina CCL11 , Citocinas/farmacologia , Relação Dose-Resposta a Droga , Endocitose/efeitos dos fármacos , Inibidores Enzimáticos/farmacologia , Eosinófilos/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Fator Estimulador de Colônias de Granulócitos e Macrófagos/farmacologia , Humanos , Interferon gama/farmacologia , Interleucinas/farmacologia , Medições Luminescentes , Camundongos , RNA Mensageiro/genética , Receptores CCR3 , Receptores de Quimiocinas/genética , Proteínas Recombinantes de Fusão/biossíntese , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fatores de Tempo , Transcrição Gênica/efeitos dos fármacos , Fator de Necrose Tumoral alfa/farmacologia , Wortmanina
16.
Eur J Immunol ; 31(8): 2458-66, 2001 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-11500830

RESUMO

The viral CC chemokine macrophage inhibitory protein-II (vMIP-II) encoded by human herpes virus 8 (HHV-8) binds to multiple chemokine receptors, however, its ability to control the initial recruitment of specific leukocyte subtypes from the peripheral circulation has not been fully clarified. Here we show that vMIP-II blocks the firm arrest and transmigration of monocytes or Th1-like T lymphocytes triggered by RANTES immobilized on activated human microvascular endothelium (HMVEC) under flow conditions. The internalization of the receptors CCR1 and CCR5 that mediate arrest and transmigration of these cells in response to RANTES was prevented by vMIP-II, supporting its role as an antagonist of CCR1 and CCR5. In contrast, vMIP-II triggered the firm arrest of eosinophils and Th2-like T cells by engaging CCR3, as confirmed by its down-regulation. Immunohistochemical analysis of HHV-8-associated Kaposi's sarcoma lesions marked by vMIP-II expression and mononuclear cell infiltration revealed a predominance of Th2-type CCR3(+) lymphocytes over Th1-type CXCR3(+)/CCR5(+) leukocytes, indicating that as a CCR3 agonist vMIP-II can drive a Th2-type immune response in vivo. Thus, our data provide evidence for a immunomodulatory role of vMIP-II in directing inflammatory cell recruitment away from a Th1-type towards a Th2-type response and thereby facilitating evasion from cytotoxic reactions.


Assuntos
Quimiocinas/imunologia , Citotoxicidade Imunológica/imunologia , Herpesvirus Humano 8/imunologia , Monócitos/imunologia , Células Th2/imunologia , Antagonistas dos Receptores CCR5 , Adesão Celular/efeitos dos fármacos , Divisão Celular/efeitos dos fármacos , Linhagem Celular , Quimiocina CCL5/antagonistas & inibidores , Quimiocina CCL5/metabolismo , Quimiocina CCL5/farmacologia , Quimiocinas/genética , Quimiocinas/farmacologia , Quimiotaxia de Leucócito/efeitos dos fármacos , Citotoxicidade Imunológica/efeitos dos fármacos , Endocitose/efeitos dos fármacos , Endotélio Vascular/citologia , Endotélio Vascular/efeitos dos fármacos , Endotélio Vascular/metabolismo , Ensaio de Imunoadsorção Enzimática , Eosinófilos/citologia , Eosinófilos/efeitos dos fármacos , Humanos , Imuno-Histoquímica , Interleucina-1/imunologia , Ativação Linfocitária/efeitos dos fármacos , Monócitos/citologia , Monócitos/efeitos dos fármacos , Receptores CCR1 , Receptores CCR3 , Receptores CCR5/metabolismo , Receptores de Quimiocinas/agonistas , Receptores de Quimiocinas/antagonistas & inibidores , Receptores de Quimiocinas/metabolismo , Sarcoma de Kaposi/imunologia , Sarcoma de Kaposi/metabolismo , Sarcoma de Kaposi/patologia , Sarcoma de Kaposi/virologia , Células Th1/citologia , Células Th1/efeitos dos fármacos , Células Th1/imunologia , Células Th2/citologia , Células Th2/efeitos dos fármacos , Células Th2/metabolismo
17.
J Biol Chem ; 276(37): 34408-18, 2001 Sep 14.
Artigo em Inglês | MEDLINE | ID: mdl-11448957

RESUMO

The CC chemokine receptor CCR5 mediates chemotaxis of leukocytes and serves as a principal co-receptor for macrophage-tropic human immunodeficiency virus type 1. To identify determinants on the CCR5 carboxyl-terminal domain that regulate receptor signaling and internalization, we generated several CCR5 mutants, which were progressively shortened from the COOH terminus or had carboxyl-terminal serine, cysteine, or leucine residues substituted by alanine and expressed them in RBL-2H3 cells. Using fluorescence resonance energy transfer between beta-arrestin and CCR5 tagged with cyan and yellow variants of green fluorescent protein, we show that high affinity association of the two molecules in living cells requires intact carboxyl-terminal serine phosphorylation sites. Phosphorylation-deficient truncation or Ser/Ala replacement mutants of CCR5 mediated a sustained calcium response and enhanced granular enzyme release in RANTES-stimulated cells. Carboxyl-terminal serine residues are critically involved in CCR5 endocytosis and a dileucine motif, similar to that implicated in the regulation of CXCR2 and CXCR4, contributes to the internalization of CCR5 in a phosphorylation-independent manner. Despite their prominent role in receptor desensitization and internalization, beta-arrestins are dispensable for the CCR5-mediated stimulation of mitogen-activated protein kinase pathways in RBL-2H3 cells. We also show that CCR5 is palmitoylated on carboxyl-terminal cysteine residues. Inhibition of CCR5 palmitoylation by alanine mutagenesis of cysteines or treatment with a palmitate analogue inhibitor profoundly reduces phorbol 12-myristate 13-acetate- and RANTES-induced receptor phosphorylation, homologous desensitization, and internalization. Alanine mutagenesis of serine, cysteine, or leucine residues or the limited carboxyl-terminal truncation of CCR5 did not impair chemokine-stimulated migration of RBL-2H3 cells. Together these results indicate that post-translational modifications of carboxyl-terminal serine and cysteine residues have a significant impact on receptor deactivation and internalization.


Assuntos
Receptores CCR5/química , Sequência de Aminoácidos , Arrestina/metabolismo , Cálcio , Linhagem Celular , Quimiocina CCL5/farmacologia , Ativação Enzimática , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Dados de Sequência Molecular , Ácido Palmítico/metabolismo , Fosforilação , Receptores CCR5/fisiologia , Relação Estrutura-Atividade , Acetato de Tetradecanoilforbol/farmacologia , Transfecção
18.
J Immunol ; 167(1): 228-34, 2001 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-11418653

RESUMO

Dendritic cells (DC) were purified by flow cytometry from rat tracheal mucosa; they exhibited the phenotypic characteristics of immature DC including high endocytic activity, low CD80/86 expression, and in vitro responsiveness to a broad range of CC chemokines. Daily treatment of adult rats with the selective CCR1 and CCR5 antagonist Met-RANTES reduced baseline numbers of tracheal intraepithelial DC by 50-60%, and pretreatment of animals with Met-RANTES before inhalation of aerosol containing heat-killed bacteria abolished the rapid DC influx into the epithelium that occurred in untreated controls, implicating CCR1 and CCR5 and their ligands in recruitment of immature DC precursors into resting airway tissues and during acute bacterial-induced inflammation. Comparable levels of DC recruitment were observed during airway mucosal Sendai virus infection and after aerosol challenge of sensitized animals with the soluble recall Ag OVA. However, Met-RANTES did not affect these latter responses, indicating the use of alternative chemokine receptors/ligands for DC recruitment, or possibly attraction of different DC subsets, depending on the nature of the eliciting stimulus.


Assuntos
Movimento Celular/imunologia , Células Dendríticas/imunologia , Células Dendríticas/patologia , Células Epiteliais/imunologia , Células Epiteliais/patologia , Interfase/imunologia , Receptores de Quimiocinas/fisiologia , Traqueia/imunologia , Traqueia/patologia , Administração por Inalação , Administração Intranasal , Aerossóis , Animais , Separação Celular , Células Dendríticas/citologia , Células Dendríticas/metabolismo , Células Epiteliais/citologia , Células Epiteliais/metabolismo , Inflamação/imunologia , Inflamação/microbiologia , Inflamação/virologia , Injeções Intraperitoneais , Moraxella catarrhalis/imunologia , Ovalbumina/administração & dosagem , Ovalbumina/imunologia , Ratos , Ratos Endogâmicos , Receptores de Quimiocinas/biossíntese , Mucosa Respiratória/citologia , Mucosa Respiratória/imunologia , Mucosa Respiratória/metabolismo , Mucosa Respiratória/patologia , Respirovirus/imunologia , Solubilidade , Fatores de Tempo , Traqueia/citologia , Traqueia/metabolismo
19.
J Immunol ; 167(1): 524-31, 2001 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-11418691

RESUMO

The understanding of the mechanisms underlying eosinophil recruitment in vivo may aid in the development of novel strategies for the treatment of allergic disorders. In this study, we investigated the role of chemokines in the cascade of events leading to eosinophil recruitment in a stem cell factor (SCF)- and leukotriene B(4) (LTB(4))-dependent allergic pleurisy model in mice. The intrapleural administration of the eosinophil-active chemokines eotaxin, RANTES, and macrophage-inflammatory protein 1alpha (MIP-1alpha) induced a time- and dose-dependent eosinophil recruitment. Pretreatment with anti-eotaxin, but not anti-RANTES or anti-MIP-1alpha, blocked the recruitment of eosinophils following Ag challenge of sensitized animals, and significant eotaxin immunoreactivity was detected in the pleural cavity of these animals. Similarly, only the anti-eotaxin inhibited the eosinophil recruitment induced by injection of SCF in naive animals. However, blockade of SCF did not inhibit the release of eotaxin after Ag challenge of sensitized mice. Akin to its effects on SCF and in the allergic reaction, eotaxin-induced eosinophil recruitment was blocked by the LTB(4) receptor antagonist CP105696. Nevertheless, SCF, but not eotaxin, appeared to regulate the endogenous release of LTB(4) after Ag challenge. Finally, we show that low doses of eotaxin synergized with LTB(4) to induce eosinophil recruitment in the pleural cavity. Overall, the present results show that eotaxin and SCF-induced LTB(4) cooperate to induce eosinophil recruitment into sites of allergic inflammation. Cooperation between inflammatory mediators must be an important phenomenon in vivo, explaining both the ability of lower concentrations of mediators to induce a full-blown functional response and the effectiveness of different strategies at inhibiting these responses.


Assuntos
Movimento Celular/imunologia , Quimiocinas CC , Citocinas/fisiologia , Leucotrieno B4/biossíntese , Pleurisia/imunologia , Hipersensibilidade Respiratória/imunologia , Fator de Células-Tronco/administração & dosagem , Animais , Quimiocina CCL11 , Quimiocina CCL3 , Quimiocina CCL4 , Quimiocina CCL5/administração & dosagem , Quimiocina CCL5/análogos & derivados , Citocinas/administração & dosagem , Injeções Intraperitoneais , Injeções Subcutâneas , Leucotrieno B4/fisiologia , Proteínas Inflamatórias de Macrófagos/administração & dosagem , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Ovalbumina/administração & dosagem , Pleura/imunologia , Pleura/patologia , Pleurisia/patologia , Receptores de Quimiocinas/antagonistas & inibidores , Hipersensibilidade Respiratória/patologia
20.
J Leukoc Biol ; 69(6): 977-85, 2001 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-11404385

RESUMO

Human RANTES (CCL5) and MIP-1alpha (CCL3) bind and activate several CC chemokine receptors. RANTES is a high-affinity ligand for CCR1 and CCR5, and it binds CCR3 with moderate affinity and CCR4 with low affinity. MIP-1alpha has similar binding characteristics to RANTES except that it does not bind to CCR3. Here we have generated a chimera of human MIP-1alpha and RANTES, called MIP/RANTES, consisting of the eight amino terminal residues of MIP-1alpha preceding the CC motif, and the remainder of the sequence is RANTES. The chimera is able to induce chemotaxis of human monocytes. MIP/RANTES has >100-fold reduction in binding to CCR1 and does not bind to CCR3 but retains full, functional binding to CCR5. It has equivalent affinity for CCR5 to MIP-1alpha and RANTES, binding with an IC(50) of 1.12 nM, and is able to mobilize calcium and induce endocytosis of CCR5 in PBMC in a manner equi-potent to RANTES. It also retains the ability to inhibit R5 using HIV-1 strains. Therefore, we conclude that the amino terminus of RANTES is not involved in CCR5 binding, but it is essential for CCR1 and CCR3.


Assuntos
Quimiocina CCL5/metabolismo , Proteínas Inflamatórias de Macrófagos/metabolismo , Receptores CCR5/metabolismo , Receptores de Quimiocinas/metabolismo , Sequência de Aminoácidos , Ligação Competitiva , Sinalização do Cálcio/efeitos dos fármacos , Quimiocina CCL3 , Quimiocina CCL4 , Quimiocina CCL5/análogos & derivados , Quimiocina CCL5/química , Quimiotaxia/efeitos dos fármacos , Regulação para Baixo/efeitos dos fármacos , HIV-1/fisiologia , Proteínas Inflamatórias de Macrófagos/química , Dados de Sequência Molecular , Monócitos/efeitos dos fármacos , Ligação Proteica , Estrutura Terciária de Proteína , Receptores CCR1 , Receptores CCR4 , Proteínas Recombinantes de Fusão/metabolismo , Proteínas Recombinantes de Fusão/fisiologia , Alinhamento de Sequência , Homologia de Sequência de Aminoácidos
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