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Proinflammatory profile of in vitro monocytes in the ageing is affected by lymphocytes presence

Pinke, Karen Henriette; Calzavara, Bruno; Faria, Patricia Freitas; Nascimento, Magda Paula Pereira do; Venturini, James; Lara, Vanessa Soares
Fonte: Biomed Central; London Publicador: Biomed Central; London
Tipo: Artigo de Revista Científica
Português
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36.84%
Background: Aging is associated with complex and constant remodeling of the immune function, resulting in an increasing susceptibility to infection and others diseases. The infections caused by Gram-negative microorganisms, present in nursing homes and hospitals, constitute one of the most common infections in the elderly, and are mainly combated by innate immune cells. Although the functions of innate immunity seem more preserved during aging than of adaptive immune mechanisms, two systems operate in an integrated way in the body, so that injury in one part of the immune system inevitably affects the other as they are part of a defensive network. The aim of this study was to investigate the in vitro production of proinflammatory (TNF-α, IL-6, IL-1α, CXCL-8 and MCP-1) and antiinflammatory (TGF-α and IL-10) cytokines by monocytes, stimulated or not (basal) with lipopolysaccharide, from healthy young and elderly subjects. By means of PBMCs, we also studied if cytokine profile is altered in these different patient groups, in the presence of lymphocytes, under the same experimental conditions. Results: The monocytes from elderly presented higher basal production of TNF-α, MCP-1 and lower of TGF-α than young monocytes. PBMC showed similar cytokines production...

Produção de citocinas pelos monócitos de idosos com estomatite protética associada a Candida; Cytokine production by monocytes from elderly patients with Candida-associated denture stomatitis

Faria, Patricia Freitas
Fonte: Biblioteca Digitais de Teses e Dissertações da USP Publicador: Biblioteca Digitais de Teses e Dissertações da USP
Tipo: Tese de Doutorado Formato: application/pdf
Publicado em 07/10/2011 Português
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Estomatite protética associada a Cândida (EPC), a lesão mais frequente em usuários de próteses removíveis, principalmente os idosos, caracteriza-se por uma inflamação da mucosa bucal que suporta a prótese. Está fortemente associada com Candida Albicans, bem como com fatores locais e sistêmicos, como a deficiência da resposta imune. Os monócitos são importantes na resposta protetora contra o fungo, produzindo citocinas que recrutam e ativam leucócitos. Existem alterações funcionais dessas células com o avanço da idade. Não foi possível encontrar na literatura dados referentes à função imunomodulatória dos monócitos de idosos com EPC. O presente trabalho pretendeu avaliar a produção de citocinas por essas células, estimuladas in vitro com C. albicans, obtidas do sangue periférico de idosos usuários de prótese total superior (PTS) com EPC, comparando-se com idosos usuários de PTS sem EPC, e com idosos e jovens não usuários de PTS. Os monócitos isolados foram cultivados em placas de cultura de 24 poços, na ausência ou presença de lipopolissacarídeo (LPS) ou C. albicans ATCC 90028 morta pelo calor. Após 18 horas, o sobrenadante foi coletado e submetido ao ensaio de imunoabsorção por ligação enzimática (ELISA) para dosagem das citocinas pró- inflamatórias fator de necrose tumoral- (TNF-)...

Estudo de biomarcadores imunológicos  das funções de monócitos derivados de pacientes HIV+; Study of immunological biomarkers of functions of monocytes derived from HIV + patients

Cacemiro, Maira da Costa
Fonte: Biblioteca Digitais de Teses e Dissertações da USP Publicador: Biblioteca Digitais de Teses e Dissertações da USP
Tipo: Dissertação de Mestrado Formato: application/pdf
Publicado em 14/02/2014 Português
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36.76%
O vírus da imunodeficiência humana (HIV) é o responsável pela pandemia mundial da síndrome da imunodeficiência adquirida (AIDS). Uma vez infectado pelo HIV, o hospedeiro apresenta a forma aguda da doença, caracterizada por aumento da carga viral circulante, rápido declínio das células TCD4+ e ativação da resposta imune inata. Quando o HIV se estabelece no organismo, induz latência em algumas células e leva à cronicidade da infecção e nessa fase o principal alvo do HIV são as células T CD4+, observa-se então constante diminuição desta população, que tem como consequência a imunodeficiência, com desativação de outras células imunes, como os monócitos, macrófagos, células Natural Killer e neutrófilos. Portanto, há o favorecimento das infecções oportunistas por fungos, bactérias, parasitas e outros vírus, além do surgimento de neoplasias principais responsáveis pela morbidade e mortalidade relacionadas à AIDS. Para conter a destruição do sistema imune pelo vírus, inicia-se a terapia antirretroviral (TARV), sendo que o parâmetro disponível na clínica para início da terapia é a carga viral e contagem de células TCD4+. Neste sentido, investigamos se fatores relacionados à função e fenótipo de monócitos de pacientes HIV+ poderiam servir como biomarcadores da progressão da infecção. Para tanto...

Aumento da expressão do receptor Toll-like 2 em monócitos do sangue periférico de pacientes com artrite psoriásica; Increased expression of Toll-like receptor 2 in peripheral blood monocytes from patients with psoriatic arthritis

Carrasco, Solange
Fonte: Biblioteca Digitais de Teses e Dissertações da USP Publicador: Biblioteca Digitais de Teses e Dissertações da USP
Tipo: Dissertação de Mestrado Formato: application/pdf
Publicado em 27/05/2014 Português
Relevância na Pesquisa
36.76%
INTRODUÇÃO: Os receptores Toll-like 2 e 4 (TLR-2 e TLR-4) são capazes de ativar células imunes inatas em resposta a bactérias Gram-positivas e Gram-negativas, respectivamente. Na artrite psoriásica (APs), doença articular inflamatória crônica, fatores genéticos, ambientais e infecciosos parecem estar envolvidos. OBJETIVO: Avaliar as expressões dos receptores: TLR-2; TLR-4; CD114 e do CD116 em monócitos e neutrófilos do sangue periférico de pacientes com APs e adicionalmente a prevalência do HLA-B27. MÉTODOS: Quarenta e cinco pacientes com diagnóstico de APs conforme os critérios CASPAR e 32 indivíduos saudáveis foram estudados. Dentre os 45 pacientes, 27 apresentavam APs ativa (DAS28 > 2,6) e 18 APs inativa (DAS28 < 2,5). A leitura das expressões do TLR-2, TLR-4, CD14, CD66, CD114, CD116 e do HLA-B27 foi realizada por citometria de fluxo no FACSCalibur da marca Becton-Dickson, utilizando anticorpos monoclonais da BD Biosciences, anti-humanos produzidos em murino. Os anticorpos monoclonais (AcMo) para marcar receptores de membrana empregados foram: CD14 conjugado com PerCP-Cy5.5 para marcar população de monócitos; CD66 conjugado com PE e FITC para população de neutrófilos; CD114 para marcar receptor de fator estimulatório de colônias de granulócitos e CD116 para marcar receptor de fator estimulatório de colônia de granulócitos-macrófagos. A análise estatística utilizou o teste U de Mann-Whitney e o teste exato de Fisher. Os valores obtidos em porcentagem foram expressos como média ± intervalo interquartil...

Production of pro- and anti-inflammatory cytokines by monocytes from patients with paracoccidioidomycosis

Peracoli, MTS; Kurokawa, C. S.; Calvi, S. A.; Mendes, R. P.; Pereira, PCM; Marques, S. A.; Soares, AMVC
Fonte: Elsevier B.V. Publicador: Elsevier B.V.
Tipo: Artigo de Revista Científica Formato: 413-418
Português
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36.79%
Monocytes and macrophages can produce a large repertoire of cytokines and participate in the pathogenesis of granulomatous diseases. We investigated the production of pro- and anti-inflammatory cytokines by monocytes from patients with active paracoccidioidomycosis. Peripheral blood monocytes from 37 patients and 29 healthy controls were cultivated with or without 10 mug/ml of lipopolysaccharide (LPS) for 18 h at 37 degreesC, and the cytokine levels were determined in the culture supernatants by enzyme immunoassay. The results showed that the endogenous levels of tumor necrosis factor alpha (TNF-alpha), interleukin-1 beta (IL-1beta), IL-6, IL-8, IL-10 and transforming growth factor beta detected in the supernatant of patient monocytes cultivated without stimulus were significantly higher than those produced by healthy controls. These data demonstrated that monocytes from patients with active paracoccidioidomycosis produce high levels of cytokines with both inflammatory and anti-inflammatory activities. However, patient monocytes produced significantly lower TNF-alpha and IL-6 levels in response to LPS when compared to normal subjects, suggesting an impairment in their capacity to produce these cytokines after LPS stimulation. Concentrations of IL-1beta...

Interleukin-15 augments oxidative metabolism and fungicidal activity of human monocytes against Paracoccidioides brasiliensis

Bannwart, Camila Ferreira; Martins, Rosana A Rodrigues; Nakaira-Takahashi, Érika; Dias-Melício, Luciane A.; Soares, Ângela MV Campos; Peraçoli, Maria Terezinha S.
Fonte: Instituto Oswaldo Cruz, Ministério da Saúde Publicador: Instituto Oswaldo Cruz, Ministério da Saúde
Tipo: Artigo de Revista Científica Formato: 866-872
Português
Relevância na Pesquisa
36.79%
Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP); Processo FAPESP: 05/54283-7; Processo FAPESP: 03/13743-0; Interleukin (IL)-15 is a pleiotropic cytokine that regulates the proliferation and survival of many cell types. IL-15 is produced by monocytes and macrophages against infectious agents and plays a pivotal role in innate and adaptive immune responses. This study analyzed the effect of IL-15 on fungicidal activity, oxidative metabolism and cytokine production by human monocytes challenged in vitro with Paracoccidioides brasiliensis (Pb18), the agent of paracoccidioidomycosis. Peripheral blood monocytes were pre-incubated with IL-15 and then challenged with Pb18. Fungicidal activity was assessed by viable fungi recovery from cultures after plating on brain-heart infusion-agar. Superoxide anion (O2-), hydrogen peroxide (H2O2), tumour necrosis factor-alpha (TNF-α), IL-6, IL-15 and IL-10 production by monocytes were also determined. IL-15 enhanced fungicidal activity against Pb18 in a dose-dependent pattern. This effect was abrogated by addition of anti-IL-15 monoclonal antibody. A significant stimulatory effect of IL-15 on O2- and H2O2 release suggests that fungicidal activity was dependent on the activation of oxidative metabolism. Pre-treatment of monocytes with IL-15 induced significantly higher levels of TNF-α...

Downregulation of nuclear factor-kappa B (NF-kappa B) pathway by silibinin in human monocytes challenged with Paracoccidioides brasiliensis

Bannwart-Castro, Camila F.; Nakaira-Takahagi, Erika; Golim, Marjorie Assis; Lopes de Medeiros, Leonardo Teixeira; Romao, Mariana; Weel, Ingrid Cristina; Serrao Peracoli, Maria Terezinha
Fonte: Pergamon-Elsevier B.V. Ltd Publicador: Pergamon-Elsevier B.V. Ltd
Tipo: Artigo de Revista Científica Formato: 880-886
Português
Relevância na Pesquisa
36.79%
Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP); Processo FAPESP: 08/08374-9; Aims: Silibinin is the major active component of silymarin, a polyphenolic plant flavonoid that has anti-inflammatory effects. The modulatory effect of silibinin on monocyte function against Paracoccidioides brasiliensis (Pb18) has not yet been demonstrated. The present study investigated whether the effect of silibinin on nuclear factor-kappa B (NF-kappa B) pathways may affect the production of tumor necrosis factor-alpha (TNF-alpha), interleukin-10 (IL-10), transforming growth factor beta (TGF-beta 1), prostaglandin E(2) (PGE(2)), nitric oxide (NO) and fungicidal activity of human monocytes challenged in vitro with Pb18.Main methods: Peripheral blood monocytes from healthy individuals were treated with silibinin and challenged with Pb18 for 18 h. TNF-alpha, IL-10, TGF-beta 1 and PGE(2) expression were determined by immunoenzymatic assay (ELISA) and NO release was determined by the accumulation of nitrite in culture supernatants. Fungicidal activity of monocytes was analyzed after treatment with interferon-gamma plus silibinin and challenge with Pb18. NF-kappa B activation in cultured monocytes was evaluated by flow cytometry and ELISA.Key findings: Silibinin partially inhibited p65NF-kappa B activation as the number of cells expressing this factor was reduced and the concentration of nuclear p65NF-kappa B was low...

Effect of prostaglandins on the production of H2O2 and cytokines that modulate the fungicidal activity of human monocytes against Paracoccidioides brasiliensis

De Campos Soares, Ângela Maria Victoriano
Fonte: Universidade Estadual Paulista Publicador: Universidade Estadual Paulista
Tipo: Artigo de Revista Científica Formato: 149
Português
Relevância na Pesquisa
36.79%
Human monocytes lack fungicidal activity against high virulent strains of Paracoccidioides brasiliensis, the etiological agent of paracoccidioidomycosis, even after IFN-γ activation. However, monocytes treated with indomethacin (INDO) or INDO plus IFN-γ effectively killed this fungus, suggesting an inhibitory role of prostaglandins in this process. Thus, the purpose of this work was to test if this regulatory effect of prostaglandin was associated with alterations on H2O2 production and/or on modulatory cytokines levels, such as TNF-α, IL-10, and IL-6. Peripheral blood monocytes obtained from 10 healthy donors were incubated for 18 hours in the presence or absence of IFN-γ, INDO, or IFN-γ plus INDO, and further challenged with a high virulent strain of P. brasiliensis (Pb18) for 4 hours. Then, the monocytes cultures were evaluated for H2O2 release and fungicidal activity calculated by counting the colony forming units after plating. Moreover, on supernatants of the same cultures, TNF-α, IL-10, IL-6, and PGE2 concentrations were evaluated by ELISA. Monocytes treated with INDO or INDO plus IFN-γ presented higher fungicidal activity associated with the release of higher levels of H2O2 and TNF-α, but lesser levels of PGE2, when compared to nontreated cells. However...

Silibinin attenuates oxidative metabolism and cytokine production by monocytes from preeclamptic women.

Cristofalo, R.; Bannwart-Castro, C. F.; Magalhães, C. G.; Borges, V. T.; Peraçoli, José Carlos; Witkin, S. S.; Peraçoli, M. T.
Fonte: Universidade Estadual Paulista Publicador: Universidade Estadual Paulista
Tipo: Artigo de Revista Científica Formato: 268-275
Português
Relevância na Pesquisa
36.79%
Silibinin is a polyphenolic plant flavonoid with anti-inflammatory properties. The present study investigated the effect of silibinin on oxidative metabolism and cytokine production - tumor necrosis factor-alpha (TNF-α), interleukin (IL)12, granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-6, IL-10, and transforming growth factor beta (TGF-β1) - by peripheral blood monocytes (PBM) from preeclamptic pregnant women. It is a case-controlled study involving women with preeclampsia (PE, n = 30) compared with normotensive pregnant (NT, n = 30) and with non-pregnant (NP, n = 30) women. Monocytes were obtained and cultured with or without silibinin (5 μM or 50 μM) for 18 h. Superoxide anion (O2-) and hydrogen peroxide (H2O2) release were determined by specific assays, and cytokine levels were determined by immunoenzymatic assays (ELISA). Monocytes from preeclamptic women cultured without stimulus released higher levels of O22, H2O2 and TNF-α, and lower levels of IL-10 and TGF-β1 than did monocytes from NT and NP women. Treatment in vitro with silibinin significantly inhibited spontaneous O2- and H2O2 release and TNF-α production by monocytes from preeclamptic women. The main effect of silibinin was obtained at 50 μM concentration. Thus...

Proinflammatory profile of in vitro monocytes in the ageing is affected by lymphocytes presence

Pinke, Karen Henriette; Calzavara, Bruno; Faria, Patricia Freitas; do Nascimento, Magda Paula Pereira; Venturini, James; Lara, Vanessa Soares
Fonte: Universidade Estadual Paulista Publicador: Universidade Estadual Paulista
Tipo: Artigo de Revista Científica
Português
Relevância na Pesquisa
36.84%
Background: Aging is associated with complex and constant remodeling of the immune function, resulting in an increasing susceptibility to infection and others diseases. The infections caused by Gram-negative microorganisms, present in nursing homes and hospitals, constitute one of the most common infections in the elderly, and are mainly combated by innate immune cells. Although the functions of innate immunity seem more preserved during aging than of adaptive immune mechanisms, two systems operate in an integrated way in the body, so that injury in one part of the immune system inevitably affects the other as they are part of a defensive network. The aim of this study was to investigate the in vitro production of proinflammatory (TNF-α, IL-6, IL-1β, CXCL-8 and MCP-1) and anti-inflammatory (TGF-β and IL-10) cytokines by monocytes, stimulated or not (basal) with lipopolysaccharide, from healthy young and elderly subjects. By means of PBMCs, we also studied if cytokine profile is altered in these different patient groups, in the presence of lymphocytes, under the same experimental conditions.Results: The monocytes from elderly presented higher basal production of TNF-α, MCP-1 and lower of TGF-β than young monocytes. PBMC showed similar cytokines production...

Effect of monocytes/macrophages on the early osteogenic differentiation of hBMSCs

Pirraco, R. P.; Reis, R. L.; Marques, A. P.
Fonte: Wiley & Sons, Inc. Publicador: Wiley & Sons, Inc.
Tipo: Artigo de Revista Científica
Publicado em /05/2013 Português
Relevância na Pesquisa
36.81%
Heterotypic cell interactions are essential for the homeostasis of bone tissue, in particular the widely studied interaction between osteoblasts and osteoclasts. Closely related with osteoclasts are monocytes/macrophages. These have been shown to produce osteogenic factors, e.g. BMP-2, which plays a key role in bone metabolism. However, the mechanisms through which monocytes/macrophages interact with osteoblasts are still elusive. The aim of this work was to assess the influence of human peripheral blood monocytes/macrophages over the early osteogenic differentiation of human bone marrow stromal cells (hBMSCs) in the presence of dexamethasone-supplemented medium. The co-cultures were performed using porous transwells that allowed the interaction between both cell types through the production of paracrine factors. The potential effect of BMP-2 produced by monocytes/macrophages was addressed by adding an anti-BMP-2 antibody to the co-cultures. hBMSCs cultured in the presence of monocytes/macrophages had a higher proliferation rate than hBMSCs monocultures. The quantification of early osteogenic marker alkaline phosphatase (ALP) revealed higher activity of this enzyme in cells in the co-culture throughout the time of culture. Both of these effects were inhibited by adding an anti-BMP-2 antibody to the cultures. Moreover...

Interleukin-15 augments oxidative metabolism and fungicidal activity of human monocytes against Paracoccidioides brasiliensis

Bannwart,Camila Ferreira; Martins,Rosana A Rodrigues; Nakaira-Takahashi,Érika; Dias-Melício,Luciane A; Soares,Ângela MV Campos; Peraçoli,Maria Terezinha S
Fonte: Instituto Oswaldo Cruz, Ministério da Saúde Publicador: Instituto Oswaldo Cruz, Ministério da Saúde
Tipo: Artigo de Revista Científica Formato: text/html
Publicado em 01/11/2010 Português
Relevância na Pesquisa
36.79%
Interleukin (IL)-15 is a pleiotropic cytokine that regulates the proliferation and survival of many cell types. IL-15 is produced by monocytes and macrophages against infectious agents and plays a pivotal role in innate and adaptive immune responses. This study analyzed the effect of IL-15 on fungicidal activity, oxidative metabolism and cytokine production by human monocytes challenged in vitro with Paracoccidioides brasiliensis (Pb18), the agent of paracoccidioidomycosis. Peripheral blood monocytes were pre-incubated with IL-15 and then challenged with Pb18. Fungicidal activity was assessed by viable fungi recovery from cultures after plating on brain-heart infusion-agar. Superoxide anion (O2-), hydrogen peroxide (H2O2), tumour necrosis factor-alpha (TNF-α), IL-6, IL-15 and IL-10 production by monocytes were also determined. IL-15 enhanced fungicidal activity against Pb18 in a dose-dependent pattern. This effect was abrogated by addition of anti-IL-15 monoclonal antibody. A significant stimulatory effect of IL-15 on O2- and H2O2 release suggests that fungicidal activity was dependent on the activation of oxidative metabolism. Pre-treatment of monocytes with IL-15 induced significantly higher levels of TNF-α, IL-10 and IL-15 production by cells challenged with the fungus. These results suggest a modulatory effect of IL-15 on pro and anti-inflammatory cytokine production...

Avaliação dos monócitos na Leishmaniose Tegumentar Americana; Evaluation of monocytes profile in American Tegumentary Leishmaniasis

PEREIRA, Ledice Inacia de Araujo
Fonte: Universidade Federal de Goiás; BR; UFG; Doutorado em Medicina Tropical; Ciências da Saúde Publicador: Universidade Federal de Goiás; BR; UFG; Doutorado em Medicina Tropical; Ciências da Saúde
Tipo: Tese de Doutorado Formato: application/pdf
Português
Relevância na Pesquisa
36.85%
American Tegumentary Leishmaniasis (ATL) is caused by Leishmania protozoan that infects mononuclear phagocytic cells leading to cutaneous or mucosal lesions. The mechanisms responsible for parasite control or persistence are not completely known. Human monocytes are blood cells subdivided into three subsets (classical, nonclassical and intermediate) according to the CD14 and CD16 expression that is associated with different phenotypical and functional characteristics. The aim of this study was to evaluate the profile of monocytes in ATL. First, it was analyzed the nonclassical monocytes (CD14loCD16+) and CD56+CD16+ NK cell frequencies in peripheral blood (by using flow cytometry) of a patient with diffuse cutaneous leishmaniasis (DCL) before, during and after immunochemotherapy (chemotherapy treatment together with L. (L.) amazonensis and L. (V.) braziliensis monovalent vaccines plus BCG). Then, in localized cutaneous leishmaniasis (LCL) patients (n = 32) and healthy donors, the three monocyte subsets (CD14hiCD16-, CD14hiCD16+, CD14loCD16+) were evaluated by flow cytometry; in the whole blood cultures we evaluated the expression of cytokines in CD14+ monocytes activated with lipopolysaccharide (LPS, by flow cytometry), and the secretion of proinflammatory (tumor necrosis factor...

AICL is a ligand for the human NK receptor NKp80/KLRF1 and mediates an activating crosstalk between NK cells and monocytes; AICL ist ein Ligand des humanen NK Rezeptors NKp80/KLRF1 und vermittelt eine aktivierende Interaktion zwischen NK-Zellen und Monozyten

Welte, Stefan
Fonte: Universidade de Tubinga Publicador: Universidade de Tubinga
Tipo: Dissertação
Português
Relevância na Pesquisa
36.85%
NK cell activity is governed by the complex interplay of multiple activating, inhibitory and co-stimulatory receptors. Therefore, a thorough understanding of NK cell biology requires the functional definition of these various receptors and their ligands. The NKC-encoded, homodimeric C-type lectin-like molecule NKp80 has been described as an activating orphan receptor, which is almost exclusively expressed by NK cells and stimulates NK cytolysis. This thesis identifies the adjacently encoded C-type lectin-like molecule AICL as a physiological ligand of NKp80. It also defines AICL as a myeloid-specific, activating receptor expressed on monocytes, macrophages and granulocytes. AICL is up-regulated in response to TLR stimulation and down-regulated during differentiation of monocytes to immature DCs. Cross-linking of AICL stimulated TNFalpha-release by monocytes, which was further enhanced by LPS stimulation. Additionally, it was shown that the NKp80-AICL interaction promotes lysis of AICL-expressing cells by NK cells. Malignant U937 cells were strongly lyzed by NK cells, whereas cytolysis of autologous LPS-activated monocytes was considerably lower or even absent depending on the donor. In other studies NK cell-mediated killing of infected monocytes and macrophages has been reported...

Phagozytose und Rezeptorexpression von Monozyten unter dem Enfluss künstlicher Kolloide und LPS in vitro; Influence of artificial colloids and LPS on phagocytosis and receptor expression of monocytes in vitro

Aberle, Dagmar Barbara
Fonte: Universidade de Tubinga Publicador: Universidade de Tubinga
Tipo: Dissertação
Português
Relevância na Pesquisa
36.87%
Gegenstand der vorliegenden Arbeit sind Untersuchungen zum Einfluss künstlicher Kolloide, Dextran, Hydroxyethylstärke (HES) und Gelatine auf die Phagozytoseleistung von Monozyten in vitro. Es wurden vergleichende Untersuchungen zwischen aus buffy coat isolierten Monozyten und Monozyten im Vollblut durchgeführt. In einem weiteren Ansatz wurden Rezeptorexpressionen von isolierten Monozyten nach Inkubation mit künstlichen Kolloiden bestimmt. Der Einfluss von Lipopolysaccharid (LPS) wurde in allen Versuchsansätzen ebenfalls untersucht. Isolierte Monozyten wurden mit Dextran, HES und Gelatine jeweils in Konzentrationen von 10, 20 und 40 mg/ml inkubiert. Unter Berücksichtigung eines Einflusses von LPS (100 ng/ml) wurde die Phagozytosekapazität der Monozyten für fluoreszenzmarkierte Latex-Beads in einer durchflusszytometrischen Analyse untersucht: Die Ergebnisse zeigen eine signifikante Einschränkung der phagozytären Leistung unter steigenden Kolloidkozentrationen, bei Inkubation mit Dextran und Gelatine bereits in klinisch relevanten Konzentrationen von 10 mg/ml. Der Einfluss von LPS ist ebenfalls signifikant und führt zu einem zusätzlichen Rückgang der Phagozytose. Bei den identischen Versuchsansätzen an Monozyten im Vollblut kam es hingegen zu einer Zunahme der phagozytären Kapazität unter steigenden Kolloidkonzentrationen. Dextran erhöht die phagozytäre Leistung von Monozyten in Konzentrationen von 10 mg/ml und 20 mg/ml sogar signifikant. Der stimulierende Einfluss der Kolloide ist auch unter LPS-Einfluss zu beobachten...

Lipopolysaccharide-Mediated Regulation of IL-17 Receptor Levels in Human Monocytes

ZHANG, Xiubo
Fonte: Quens University Publicador: Quens University
Tipo: Tese de Doutorado
Português
Relevância na Pesquisa
36.81%
IL-17 promotes inflammation through the recruitment of monocytes and induction of various chemokines and inflammatory cytokines. Monocytes respond to IL-17 through the heteromeric IL-17 receptor (IL-17R) composed of subunits IL-17RA and IL-17RC. Together, monocytes and IL-17 amplify inflammation. Controlling the cellular response to IL-17 is crucial to prevent hyperactivation of inflammatory responses, which could lead to chronic inflammatory diseases. The cellular response to increased IL-17 levels may be limited by controlling the receptor levels. Before we understand how monocytes respond to IL-17 during infection, we must first characterize the expression of IL-17R in these cells in response to LPS, a well-characterized pro-inflammatory signal. The aim of this study is to understand the mechanisms which regulate IL-17R levels in human monocytes. IL-17R mRNA and protein levels were measured in response to LPS by RT-PCR and Western blot analysis in primary human monocytes, peripheral blood mononuclear cells (PBMC), and the human monocytic cell line, THP-1. LPS enhanced IL-17RA and RC transcript levels in monocytes and PBMC. In contrast, IL-17RA protein levels decreased with LPS treatment in these cells. Investigation into mechanisms regulating IL-17RA protein levels lead to the observation that IL-17RA undergoes receptor degradation in response to LPS. This work identifies for the first time that 1) LPS enhances transcript levels of IL-17R and 2) after LPS treatment...

Analise por citometria de fluxo da expressão dos receptores FCy e complemento em monocitos de pacientes com agamaglobulinemia ligada ao X e imunodeficiencia comum variavel; Flow cytometry analysis of the expression of FCy and complement receptors in monocytes of X-linked agammaglobulinaemia and common variable immunodeficiency patients

Ana Lidia Braga Amoras
Fonte: Biblioteca Digital da Unicamp Publicador: Biblioteca Digital da Unicamp
Tipo: Tese de Doutorado Formato: application/pdf
Publicado em 18/12/2009 Português
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36.79%
Recentemente, demonstramos defeitos na fagocitose e quimiotaxia de monócitos em pacientes com agamaglobulinemia ligada ao X (ALX) e imunodeficiência comum variável (ICV). Existem poucos dados da expressão in vivo dos receptores para a região constante de imunoglobulina G (IgG) (FcyR) e receptores de complemento (RC) nesses pacientes. O objetivo deste estudo foi investigar a expressão dos FcyR e RC nos monócitos de pacientes com ALX e ICV e compará-la com controles saudáveis. Amostras de sangue total foram obtidas de 10 pacientes com ALX, 12 com ICV e 18 controles saudáveis. O fenótipo dos monócitos foi determinado por citometria de fluxo de células marcadas com CD14+. A expressão dos receptores de superfície FcyRI (CD64), FcyRII (CD32) e FcyRIII (CD16), CR1 (CD35) e CR3 (CD11b e CD18) foi medida através da determinação da proporção de células CD14+ para cada receptor e pela densidade do receptor. Comparado ao controle, observou-se uma porcentagem significantemente maior de monócitos CD16+ e CD35+ nos pacientes com ALX (P=0,002 e P=0,007, respectivamente). A expressão da intensidade de fluorescência relativa (IFR) do FcyRII (CD32) e FcyRIII (CD16) foi significantemente mais baixa nos monócitos de pacientes com ICV em comparação aos controles (P=0...

Lipopolysaccharide-induced expression of cell surface receptors and cell activation of neutrophils and monocytes in whole human blood

Gomes,N.E.; Brunialti,M.K.C.; Mendes,M.E.; Freudenberg,M.; Galanos,C.; Salomão,R.
Fonte: Associação Brasileira de Divulgação Científica Publicador: Associação Brasileira de Divulgação Científica
Tipo: Artigo de Revista Científica Formato: text/html
Publicado em 01/09/2010 Português
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Lipopolysaccharide (LPS) activates neutrophils and monocytes, inducing a wide array of biological activities. LPS rough (R) and smooth (S) forms signal through Toll-like receptor 4 (TLR4), but differ in their requirement for CD14. Since the R-form LPS can interact with TLR4 independent of CD14 and the differential expression of CD14 on neutrophils and monocytes, we used the S-form LPS from Salmonella abortus equi and the R-form LPS from Salmonella minnesota mutants to evaluate LPS-induced activation of human neutrophils and monocytes in whole blood from healthy volunteers. Expression of cell surface receptors and reactive oxygen species (ROS) and nitric oxide (NO) generation were measured by flow cytometry in whole blood monocytes and neutrophils. The oxidative burst was quantified by measuring the oxidation of 2',7'-dichlorofluorescein diacetate and the NO production was quantified by measuring the oxidation of 4-amino-5-methylamino-2',7'-difluorofluorescein diacetate. A small increase of TLR4 expression by monocytes was observed after 6 h of LPS stimulation. Monocyte CD14 modulation by LPS was biphasic, with an initial 30% increase followed by a 40% decrease in expression after 6 h of incubation. Expression of CD11b was rapidly up-regulated...

Ratio of monocytes to lymphocytes in peripheral blood in patients diagnosed with active tuberculosis

Wang,Jun; Yin,Yongmei; Wang,Xuedong; Pei,Hao; Kuai,Shougang; Gu,Lan; Xing,Huiqin; Zhang,Yu; Huang,Qiusheng; Guan,Bin
Fonte: Brazilian Society of Infectious Diseases Publicador: Brazilian Society of Infectious Diseases
Tipo: Artigo de Revista Científica Formato: text/html
Publicado em 01/04/2015 Português
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Objective: The ratio of monocytes to lymphocytes in peripheral blood could reflect an indi- vidual's immunity to Mycobacterium tuberculosis. The objective of this study was to evaluate the relationship between ratio of monocytes to lymphocytes and clinical status of patients with active tuberculosis. Methods: This was a retrospective review of data collected from the clinical database of The Fifth People's Hospital of Wuxi, Medical College of Jiangnan University. A total of 419 patients who had newly diagnosed active tuberculosis and 108 cases from 419 patients with tuberculosis therapy either near completion or completed were selected. Controls were 327 healthy donors. Results: Median ratio of monocytes to lymphocytes was 0.36 (IQR, 0.22-0.54) in patients before treatment, and 0.16 (IQR, 0.12-0.20) in controls (p < 0.001). Ratio of monocytes to lymphocytes <9% or >25% was significant predictors for active tuberculosis (OR = 114.73, 95% CI, 39.80-330.71; OR = 89.81, 95% CI, 53.18-151.68, respectively). After treatment, the median ratio of monocytes to lymphocytes recovered to be nearly normal. Compared to other patients, patients with extrapulmonary tuberculosis and of age >60 years were more likely to have extreme ratio of monocytes to lymphocytes (AOR = 2.57...

Mechanisms of cytokine release induced by electronegative LDL in monocytes. The role of ceramide and the receptors CD14-TLR4

Estruch Alrich, Montserrat
Fonte: [Barcelona] : Universitat Autònoma de Barcelona, Publicador: [Barcelona] : Universitat Autònoma de Barcelona,
Tipo: Tesis i dissertacions electròniques; info:eu-repo/semantics/doctoralThesis; info:eu-repo/semantics/publishedVersion Formato: application/pdf
Publicado em //2014 Português
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L'LDL electronegativa (LDL(-)) és una LDL modificada present en circulació amb propietats inflamatòries incloent la inducció de l'alliberament de citoquines en cèl·lules relacionades amb l'arteriosclerosi, tals com monòcits i cèl·lules endotelials. Es coneix poc sobre les vies intracel·lulars activades per l'LDL(-), en especial en monòcits. En aquesta tesi s'han estudiat els primers passos a través dels quals l'LDL(-) indueix l'alliberament d'IL-6, IL-10 i MCP-1 en monòcits humans. Concretament, s'han estudiat els components inflamatoris de l'LDL(-) i la seva interacció amb receptors cel·lulars de monòcits. En relació amb els components inflamatoris, es va observar que la ceramida (CER) és, en part, responsable dels efectes inflamatoris de l'LDL(-). La CER es relaciona amb un augment en la susceptibilitat a l'agregació de l'LDL(-) i a la seva inducció de citoquines en monòcits. Aquestes propietats de l'LDL(-) augmenten a 37ºC i disminueixen per la incubació d'aquesta lipoproteina amb l'HDL. El contingut en CER en l'LDL(-) augmenta per acció de l'activitat tipus fosfolipasa C (PLC), intrínseca en l'LDL(-) i que també es veu inhibida per l'HDL. Aquest fet suggereix que l'activitat PLC de l'LDL(-) participa en les modificacions que tenen lloc en l'LDL nativa (LDL(+)) per a la formació de LDL(-). L'enriquiment d'LDL en CER (CER-LDL)) i el tractament d'LDL amb PLC promouen una LDL que mimetitza l'alliberament de citoquines de l'LDL(-). Tanmateix...