RESUMO
Interleukin (IL)-33 is an important cytokine in the tumour microenvironment; it is known to promote the growth and metastasis of solid cancers, such as gastric, colorectal, ovarian and breast cancer. Our group demonstrated that the IL-33/ST2 pathway enhances the development of squamous cell carcinoma (SCC). Conversely, other researchers have reported that IL-33 inhibits tumour progression. In addition, the crosstalk between IL-33, cancer cells and immune cells in SCC remains unknown. The aim of this study was to investigate the effect of IL-33 on the biology of head and neck SCC lines and to evaluate the impact of IL-33 neutralisation on the T cell response in a preclinical model of SCC. First, we identified epithelial and peritumoural cells as a major local source of IL-33 in human SCC samples. Next, in vitro experiments demonstrated that the addition of IL-33 significantly increased the proliferative index, motility and invasiveness of SCC-25 cells, and downregulated MYC gene expression in SCC cell lines. Finally, IL-33 blockade significantly delayed SCC growth and led to a marked decrease in the severity of skin lesions. Importantly, anti-IL-33 monoclonal antibody therapy increase the percentage of CD4+IFNγ+ T cells and decreased CD4+ and CD8+ T cells secreting IL-4 in tumour-draining lymph nodes. Together, these data suggest that the IL-33/ST2 pathway may be involved in the crosstalk between the tumour and immune cells by modulating the phenotype of head and neck SCC and T cell activity. IL-33 neutralisation may offer a novel therapeutic strategy for SCC.
Assuntos
Carcinoma de Células Escamosas , Movimento Celular , Proliferação de Células , Interleucina-33 , Ativação Linfocitária , Interleucina-33/metabolismo , Humanos , Carcinoma de Células Escamosas/patologia , Carcinoma de Células Escamosas/imunologia , Carcinoma de Células Escamosas/metabolismo , Animais , Ativação Linfocitária/imunologia , Invasividade Neoplásica , Camundongos , Linhagem Celular Tumoral , Neoplasias de Cabeça e Pescoço/imunologia , Neoplasias de Cabeça e Pescoço/patologia , Neoplasias de Cabeça e Pescoço/metabolismo , Linfócitos T/imunologia , Linfócitos T/metabolismo , Microambiente Tumoral/imunologia , FemininoRESUMO
Interleukin (IL)-33 is an important cytokine in the tumour microenvironment; it is known to promote the growth and metastasis of solid cancers, such as gastric, colorectal, ovarian and breast cancer. Our group demonstrated that the IL-33/ST2 pathway enhances the development of squamous cell carcinoma (SCC). Conversely, other researchers have reported that IL-33 inhibits tumour progression. In addition, the crosstalk between IL-33, cancer cells and immune cells in SCC remains unknown. The aim of this study was to investigate the effect of IL-33 on the biology of head and neck SCC lines and to evaluate the impact of IL-33 neutralisation on the T cell response in a preclinical model of SCC. First, we identified epithelial and peritumoural cells as a major local source of IL-33 in human SCC samples. Next, in vitro experiments demonstrated that the addition of IL-33 significantly increased the proliferative index, motility and invasiveness of SCC-25 cells, and downregulated MYC gene expression in SCC cell lines. Finally, IL-33 blockade significantly delayed SCC growth and led to a marked decrease in the severity of skin lesions. Importantly, anti-IL-33 monoclonal antibody therapy increase the percentage of CD4+IFNγ+ T cells and decreased CD4+ and CD8+ T cells secreting IL-4 in tumour-draining lymph nodes. Together, these data suggest that the IL-33/ST2 pathway may be involved in the crosstalk between the tumour and immune cells by modulating the phenotype of head and neck SCC and T cell activity. IL-33 neutralisation may offer a novel therapeutic strategy for SCC.
Assuntos
Animais , Camundongos , Carcinoma de Células Escamosas/imunologia , Carcinoma de Células Escamosas/patologia , Movimento Celular , Proliferação de Células , Ativação Linfocitária/imunologia , Carcinoma de Células Escamosas/metabolismo , Linfócitos T , Linhagem Celular Tumoral , Interleucina-33/metabolismo , Neoplasias de Cabeça e PescoçoRESUMO
Este estudio, tuvo como objetivo, analizar la contaminación de la arena de plazas de recreación de las escuelas municipales de educación infantil, del municipio de Pelotas, RS, Brasil. Se recolectar muestras de arena de 20 escuelas, provenientes de cuatro barrios. Las muestras fueron procesadas por el método de centrifugado-flotación. Muestras de 11 escuelas, 55% del total, fueron positivas para parásitos con potencial zoonótico. Ancylostoma fue el género más prevalente, siendo encontrado en nueve escuelas, el 45% del total. En los barrios, lo que presentó mayor contaminación, fue el Fragata, con 80,0% (4/5 escuelas), el barrio Tres Ventas, no presentó muestras positivas, siendo de esta forma, el que presentó menor porcentaje de contaminación. La aparición de parásitos, indican que las medidas preventivas deben ser adoptadas, impidiendo el acceso de animales al espacio interno de las escuelas, evitando la contaminación de la arena y demás lugares por huevos y larvas de parásitos con potencial zoonótico, como los encontrados en este estudio y así disminuir la exposición de niños y demás personas a estos agentes.
This study, had as objective, to analyze the contamination of the sand of recreation squares of the municipal schools of pre-school education, in the municipality of Pelotas, RS, Brazil. Sand samples were collected from 20 schools from four neighborhoods. The samples were processed by the centrifugal-flotation method. Samples from 11 schools, 55% of the total, were positive for parasites with zoonotic potential. Ancylostoma was the most prevalent genus, being found in nine schools, 45% of the total. Among the neighborhoods, the one that presented the highest contamination, was the Fragata, with 80,0% (4/5), the Três Vendas neighborhood, did not present positive samples, being, in this way, the one that presented smaller percentage of contamination. The occurrence of parasites indicates that preventive measures should be adopted, preventing the access of animals to the internal space of schools, avoiding the contamination of sand and other sites by eggs and larvae of parasites with zoonotic potential, as those found in this study and thus reduce the exposure of children and others to these agents.
Este estudo, teve como objetivo, analisar a contaminação da areia de praças de recreação das escolas municipais de educação infantil, do município de Pelotas, RS, Brasil. Foram coletadas amostras de areia de 20 escolas, provenientes de quatro bairros. As amostras foram processadas pelo método de centrifugo-flutuação. Amostras de 11 escolas, 55% do total, foram positivas para parasitos com potencial zoonótico. Ancylostoma foi o gênero mais prevalente, sendo encontrado em nove escolas, 45% do total. Dentre os bairros, o que apresentou maior contaminação, foi o Fragata, com 80,0% (4/5 escolas), o bairro Três Vendas, não apresentou amostras positivas, sendo desta forma, o que apresentou menor percentual de contaminação. A ocorrência de parasitos, indicam que medidas preventivas devem ser adotadas, impedindo o acesso de animais ao espaço interno das escolas, evitando a contaminação da areia e demais locais por ovos e larvas de parasitos com potencial zoonótico, como os encontrados neste estudo e assim diminuir a exposição de crianças e demais pessoas a estes agentes.
Assuntos
Humanos , Animais , Criança , Areia/análise , Parasitos , Parques Recreativos , Saneamento de Escolas , Áreas Verdes/análise , Brasil , ZoonosesRESUMO
Este estudio, tuvo como objetivo, analizar la contaminación de la arena de plazas de recreación de las escuelas municipales de educación infantil, del municipio de Pelotas, RS, Brasil. Se recolectar muestras de arena de 20 escuelas, provenientes de cuatro barrios. Las muestras fueron procesadas por el método de centrifugado-flotación. Muestras de 11 escuelas, 55% del total, fueron positivas para parásitos con potencial zoonótico. Ancylostoma fue el género más prevalente, siendo encontrado en nueve escuelas, el 45% del total. En los barrios, lo que presentó mayor contaminación, fue el Fragata, con 80,0% (4/5 escuelas), el barrio Tres Ventas, no presentó muestras positivas, siendo de esta forma, el que presentó menor porcentaje de contaminación. La aparición de parásitos, indican que las medidas preventivas deben ser adoptadas, impidiendo el acceso de animales al espacio interno de las escuelas, evitando la contaminación de la arena y demás lugares por huevos y larvas de parásitos con potencial zoonótico, como los encontrados en este estudio y así disminuir la exposición de niños y demás personas a estos agentes.(AU)
This study, had as objective, to analyze the contamination of the sand of recreation squares of the municipal schools of pre-school education, in the municipality of Pelotas, RS, Brazil. Sand samples were collected from 20 schools from four neighborhoods. The samples were processed by the centrifugal-flotation method. Samples from 11 schools, 55% of the total, were positive for parasites with zoonotic potential. Ancylostoma was the most prevalent genus, being found in nine schools, 45% of the total. Among the neighborhoods, the one that presented the highest contamination, was the Fragata, with 80,0% (4/5), the Três Vendas neighborhood, did not present positive samples, being, in this way, the one that presented smaller percentage of contamination. The occurrence of parasites indicates that preventive measures should be adopted, preventing the access of animals to the internal space of schools, avoiding the contamination of sand and other sites by eggs and larvae of parasites with zoonotic potential, as those found in this study and thus reduce the exposure of children and others to these agents.(AU)
Este estudo, teve como objetivo, analisar a contaminação da areia de praças de recreação das escolas municipais de educação infantil, do município de Pelotas, RS, Brasil. Foram coletadas amostras de areia de 20 escolas, provenientes de quatro bairros. As amostras foram processadas pelo método de centrifugo-flutuação. Amostras de 11 escolas, 55% do total, foram positivas para parasitos com potencial zoonótico. Ancylostoma foi o gênero mais prevalente, sendo encontrado em nove escolas, 45% do total. Dentre os bairros, o que apresentou maior contaminação, foi o Fragata, com 80,0% (4/5 escolas), o bairro Três Vendas, não apresentou amostras positivas, sendo desta forma, o que apresentou menor percentual de contaminação. A ocorrência de parasitos, indicam que medidas preventivas devem ser adotadas, impedindo o acesso de animais ao espaço interno das escolas, evitando a contaminação da areia e demais locais por ovos e larvas de parasitos com potencial zoonótico, como os encontrados neste estudo e assim diminuir a exposição de crianças e demais pessoas a estes agentes.(AU)
Assuntos
Humanos , Animais , Criança , Areia/análise , Áreas Verdes/análise , Parques Recreativos , Saneamento de Escolas , Parasitos , Brasil , ZoonosesRESUMO
Recent evidence indicates that nonprofessional immune cells such as epithelial cells, endothelial cells, and fibroblasts also contribute to innate immunity via secretion of cytokines. Fibroblasts are the principal type of cell found in the periodontal connective tissues and they are involved in the immune response during periodontal disease. The role of fibroblasts in the recognition of pathogens via Toll-like receptors (TLRs) has been established; however, few studies have been conducted concerning the involvement of innate immune receptors in the recognition of Candida albicans by gingival fibroblast. In the current study, we investigate the functional activity of TLR2, cluster of differentiation 14 (CD14), and myeloid differentiation primary response gene 88 (MyD88) molecules in the recognition of C. albicans by gingival fibroblast. First, we identified that gingival fibroblasts expressed TLR2, TLR3, and TLR4. Our results showed that TLR agonists had no effect on these receptors' expression by TLR2, MyD88, and CD14-deficient cells. Notably, C. albicans and a synthetic triacylated lipoprotein (Pam3CSK4) induced a remarkable increase of TLR3 expression on MyD88-deficient gingival fibroblasts. TLR4 expression levels were lower than TLR2 and TLR3 levels and remained unchanged after TLR agonist stimulation. Gingival fibroblasts presented morphological similarities; however, TLR2 deficiency on these cells leads to a lower proliferative response, whereas the deficiency on CD14 expression resulted in lower levels of type I collagen by these cells. In addition, the recognition of C. albicans by gingival fibroblasts had an effect on the secretion of cytokines and it was dependent on a specific recognition molecule. Specifically, tumor necrosis factor-α (TNF-α) production after the recognition of C. albicans was dependent on MyD88, CD14, and TLR2 molecules, whereas the production of interleukin-1ß (IL-1ß) and IL-13 was dependent on TLR2. These findings are the first to describe a role of gingival fibroblast in the recognition of C. albicans and the pathways involved in this process. An understanding of these pathways may lead to alternative treatments for patients with periodontal disease.
Assuntos
Candida albicans/metabolismo , Fibroblastos/microbiologia , Gengiva/microbiologia , Receptores de Lipopolissacarídeos/metabolismo , Fator 88 de Diferenciação Mieloide/metabolismo , Receptor 2 Toll-Like/metabolismo , Receptor 4 Toll-Like/metabolismo , Actinas/metabolismo , Animais , Células Cultivadas , Colágeno/metabolismo , Citocinas/metabolismo , Fibroblastos/metabolismo , Imunidade Inata , Mediadores da Inflamação/metabolismo , Camundongos Endogâmicos C57BL , Camundongos KnockoutRESUMO
Adhesive interactions play important roles in coordinating T cell migration and activation, which are mediated by binding of integrins to RGD motif found on extracellular matrix proteins. Disintegrins, isolated from snake venoms, contain the RGD sequence that confers selectivity to integrin interaction. We have investigated the ability of three RGD-disintegrins, ligands of alpha(5)beta(1) and alpha(v)beta(3), Flavoridin (Fl), Kistrin (Kr) and Echistatin (Ech), in modulating the activation of human T lymphocyte. The disintegrins induced T cell proliferation and CD69 expression. This activation parallels with actin cytoskeleton reorganization and tyrosine phosphorylation. Furthermore, the peptides induced focal adhesion kinase (FAK) and phosphoinositide 3-kinase (PI3K) activation. Finally, RGD-disintegrins were capable of driving NF-kappaB nuclear translocation and c-Fos expression, in a PI3K and ERK1/2 activities dependent manner. This report is the first to show that RGD-disintegrins interact with integrins on human T lymphocyte surface, modulating cell proliferation and activation of specific pathways coupled to integrin receptor.
Assuntos
Desintegrinas/farmacologia , Integrinas/metabolismo , Ativação Linfocitária/efeitos dos fármacos , Oligopeptídeos/farmacologia , Transdução de Sinais/efeitos dos fármacos , Linfócitos T/efeitos dos fármacos , Actinas/metabolismo , Animais , Antígenos CD/imunologia , Antígenos de Diferenciação de Linfócitos T/imunologia , Núcleo Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Citoesqueleto/efeitos dos fármacos , Ativação Enzimática/efeitos dos fármacos , Proteína-Tirosina Quinases de Adesão Focal/metabolismo , Humanos , Lectinas Tipo C , Ativação Linfocitária/imunologia , NF-kappa B/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo , Fosfotirosina/metabolismo , Ligação Proteica/efeitos dos fármacos , Transporte Proteico/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-fos/metabolismo , Linfócitos T/citologia , Linfócitos T/enzimologia , Linfócitos T/imunologiaRESUMO
The effects of jarastatin (JT), a monomeric RGD-disintegrin, were compared with those of the heterodimeric MLD-disintegrin, EC3, on human neutrophil activation and functions. Both disintegrins inhibited neutrophil chemotaxis induced by fMet-Leu-Phe and were also potent chemotactic agents. These effects were accompanied by an increase in actin polymerization, and both were inhibited by genistein, a tyrosine kinase inhibitor. While JT, but not other RGD-disintegrins, inhibited EC3-induced chemotaxis, EC3 was not able to inhibit JT effect. The chemotactic effect of JT was blocked by anti-alpha(M) antibody whereas anti-alpha(9)beta(1) inhibited EC3 effect. Both JT and EC3 induced focal adhesion kinase (FAK) and phosphoinositide 3-kinase (PI3K) activation. Accordingly, LY294002, a PI3K inhibitor, impaired their chemotactic effect on neutrophils. JT induced Erk-2 translocation to nucleus and a delay of the spontaneous apoptosis of neutrophils in vitro. In contrast, EC3 inhibited Erk-2 activation and had a proapoptotic effect. These effects were reverted by PD98059, an MEK 1/2 inhibitor and blocked by z-VAD-FMK, a caspase inhibitor. In addition, JT, but not EC3, increased the IL-8 mRNA levels in neutrophils. The data indicate that JT and EC3 directly activate an integrin-coupled signaling and modulate the MAPK pathway in different ways, leading the neutrophils to express different functional response.
Assuntos
Membrana Celular/metabolismo , Quimiotaxia de Leucócito/efeitos dos fármacos , Desintegrinas/farmacologia , Integrinas/efeitos dos fármacos , Neutrófilos/efeitos dos fármacos , Venenos de Víboras/farmacologia , Transporte Ativo do Núcleo Celular/efeitos dos fármacos , Transporte Ativo do Núcleo Celular/fisiologia , Animais , Anticorpos/farmacologia , Apoptose/efeitos dos fármacos , Apoptose/fisiologia , Extratos Celulares , Membrana Celular/efeitos dos fármacos , Células Cultivadas , Quimiotaxia de Leucócito/fisiologia , Desintegrinas/antagonistas & inibidores , Interações Medicamentosas/fisiologia , Inibidores Enzimáticos/farmacologia , Quinase 1 de Adesão Focal , Proteína-Tirosina Quinases de Adesão Focal , Regulação da Expressão Gênica/efeitos dos fármacos , Regulação da Expressão Gênica/fisiologia , Humanos , Integrinas/metabolismo , Interleucina-8/genética , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Sistema de Sinalização das MAP Quinases/fisiologia , Proteína Quinase 1 Ativada por Mitógeno/efeitos dos fármacos , Proteína Quinase 1 Ativada por Mitógeno/metabolismo , Neutrófilos/metabolismo , Fragmentos de Peptídeos/farmacologia , Fosfatidilinositol 3-Quinases/efeitos dos fármacos , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Tirosina Quinases/efeitos dos fármacos , Proteínas Tirosina Quinases/metabolismo , RNA Mensageiro/efeitos dos fármacos , RNA Mensageiro/metabolismoRESUMO
Recently, a new protein containing a disintegrin domain, alternagin-C (Alt-C), was purified from Bothrops alternatus venom. Unlike other disintegrins, in Alt-C an ECD amino acid mogif takes the place of the RGD sequence. Most disintegrins contain an RGD/KGD sequence and are very potent inhibitors of platelet aggregation, as well as other cell interactions with the extracellular matrix, including tumor cell metastasis and angiogenesis. The present study investigated the effects of Alt-C on human neutrophil chemotaxis in vitro and the activation of integrin-mediated pathways. Alt-C showed a potent chemotactic effect for human neutrophils when compared to N-formyl-methionyl-leucyl-phenylalanine peptide (fMLP), a classic chemotactic agent. Moreover, preincubation of neutrophils with Alt-C significantly inhibited chemotaxis toward fMLP and itself. In addition, a peptide containing an ECD sequence presented a chemotactic activity and significantly inhibited chemotaxis induced by Alt-C and fMLP. A significant increase of F-actin content was observed in cells treated with Alt-C, showing that the chemotactic activity of Alt-C on neutrophils is driven by actin cytoskeleton dynamic changes. Furthermore, this protein was able to induce an increase of phosphotyrosine content triggering focal adhesion kinase activation and its association with phosphatidylinositol 3-kinase. Alt-C was also able to induce a significant increase in extracellular signal-regulated kinase 2 nuclear translocation. The chemotactic activity of Alt-C was partially inhibited by LY294002, a specific phosphatidylinositol 3-kinase inhibitor, and by PD98056, a Map kinase kinase inhibitor. These findings suggest that Alt-C can trigger human neutrophil chemotaxis modulated by intracellular signals characteristic of integrin-activated pathways and that these effects could be related to the ECD mogif present in disintegrin-like domain.
Assuntos
Desintegrinas/química , Desintegrinas/fisiologia , Integrinas/metabolismo , Neutrófilos/citologia , Transdução de Sinais , Actinas/metabolismo , Transporte Ativo do Núcleo Celular , Motivos de Aminoácidos , Animais , Bothrops/metabolismo , Movimento Celular , Núcleo Celular/metabolismo , Quimiotaxia , Citoesqueleto/metabolismo , Relação Dose-Resposta a Droga , Inibidores Enzimáticos/farmacologia , Matriz Extracelular/metabolismo , Flavonoides/farmacologia , Humanos , Immunoblotting , Imuno-Histoquímica , Proteína Quinase 1 Ativada por Mitógeno/metabolismo , N-Formilmetionina Leucil-Fenilalanina/química , Metástase Neoplásica , Neovascularização Patológica , Neutrófilos/metabolismo , Peptídeos/química , Fosfatidilinositol 3-Quinases/metabolismo , Fosfotirosina/metabolismo , Testes de Precipitina , Estrutura Terciária de Proteína , Venenos de Serpentes/metabolismo , Fatores de TempoRESUMO
A autora procura demonstrar a importância de Melanie Klein no desenvolvimento e nos progressos da Psicanálise e em sua atividade como pioneira na Psicanálise da criança, assinalando suas principais contribuições teóricas e técnicas à Psicanálise