Differential macrophage activation alters the expression profile of NTPDase and Ecto-5´-nucleotidase
Autor(a) principal: | |
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Data de Publicação: | 2012 |
Outros Autores: | , , , , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UFRGS |
Texto Completo: | http://hdl.handle.net/10183/225270 |
Resumo: | Macrophages are key elements in the inflammatory process, whereas depending on the micro-environmental stimulation they exhibit a pro-inflammatory (classical/M1) or an anti-inflammatory/reparatory (alternative/M2) phenotype. Extracellular ATP can act as a danger signal whereas adenosine generally serves as a negative feedback mechanism to limit inflammation. The local increase in nucleotides communication is controlled by ectonucleotidases, such as members of the ectonucleoside triphosphate diphosphohydrolase (E-NTPDase) family and ecto-59-nucleotidase/CD73 (ecto-59-NT). In the present work we evaluated the presence of these enzymes in resident mice M1 (macrophages stimulated with LPS), and M2 (macrophages stimulated with IL-4) macrophages. Macrophages were collected by a lavage of the mice (6–8 weeks) peritoneal cavity and treated for 24 h with IL-4 (10 ng/mL) or LPS (10 ng/mL). Nitrite concentrations were measured using the Greiss reaction. Supernatants were harvested to determine cytokines and the ATPase, ADPase and AMPase activities were determined by the malachite green method and HPLC analysis. The expression of selected surface proteins was evaluated by flow cytometry. The results reveal that M1 macrophages presented a decreased ATP and AMP hydrolysis in agreement with a decrease in NTPDase1, -3 and ecto-59-nucleotidase expression compared to M2. In contrast, M2 macrophages showed a higher ATP and AMP hydrolysis and increased NTPDase1, -3 and ecto-59-nucleotidase expression compared to M1 macrophages. Therefore, macrophages of the M1 phenotype lead to an accumulation of ATP while macrophages of the M2 phenotype may rapidly convert ATP to adenosine. The results also showed that P1 and P2 purinoreceptors present the same mRNA profile in both phenotypes. In addition, M2 macrophages, which have a higher ATPase activity, were less sensitive to cell death. In conclusion, these changes in ectoenzyme activities might allow macrophages to adjust the outcome of the extracellular purinergic cascade in order to fine-tune their functions during the inflammatory set. |
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Zanin, Rafael FernandesBraganhol, ElizandraBergamin, Letícia ScusselCampesato, Luis Felipe IngrassiaZanotto Filho, AlfeuMoreira, Jose Claudio FonsecaMorrone, Fernanda BuenoSevigny, JeanSchetinger, Maria Rosa ChitolinaWyse, Angela Terezinha de SouzaBattastini, Ana Maria Oliveira2021-08-06T04:41:27Z20121932-6203http://hdl.handle.net/10183/225270000822638Macrophages are key elements in the inflammatory process, whereas depending on the micro-environmental stimulation they exhibit a pro-inflammatory (classical/M1) or an anti-inflammatory/reparatory (alternative/M2) phenotype. Extracellular ATP can act as a danger signal whereas adenosine generally serves as a negative feedback mechanism to limit inflammation. The local increase in nucleotides communication is controlled by ectonucleotidases, such as members of the ectonucleoside triphosphate diphosphohydrolase (E-NTPDase) family and ecto-59-nucleotidase/CD73 (ecto-59-NT). In the present work we evaluated the presence of these enzymes in resident mice M1 (macrophages stimulated with LPS), and M2 (macrophages stimulated with IL-4) macrophages. Macrophages were collected by a lavage of the mice (6–8 weeks) peritoneal cavity and treated for 24 h with IL-4 (10 ng/mL) or LPS (10 ng/mL). Nitrite concentrations were measured using the Greiss reaction. Supernatants were harvested to determine cytokines and the ATPase, ADPase and AMPase activities were determined by the malachite green method and HPLC analysis. The expression of selected surface proteins was evaluated by flow cytometry. The results reveal that M1 macrophages presented a decreased ATP and AMP hydrolysis in agreement with a decrease in NTPDase1, -3 and ecto-59-nucleotidase expression compared to M2. In contrast, M2 macrophages showed a higher ATP and AMP hydrolysis and increased NTPDase1, -3 and ecto-59-nucleotidase expression compared to M1 macrophages. Therefore, macrophages of the M1 phenotype lead to an accumulation of ATP while macrophages of the M2 phenotype may rapidly convert ATP to adenosine. The results also showed that P1 and P2 purinoreceptors present the same mRNA profile in both phenotypes. In addition, M2 macrophages, which have a higher ATPase activity, were less sensitive to cell death. In conclusion, these changes in ectoenzyme activities might allow macrophages to adjust the outcome of the extracellular purinergic cascade in order to fine-tune their functions during the inflammatory set.application/pdfengPLoS ONE. San Francisco. Vol. 7, no. 2 (Feb. 2012), e31205, 10 f.5'-nucleotidaseAtivação de macrófagosRNA mensageiroDifferential macrophage activation alters the expression profile of NTPDase and Ecto-5´-nucleotidaseEstrangeiroinfo:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFRGSinstname:Universidade Federal do Rio Grande do Sul (UFRGS)instacron:UFRGSTEXT000822638.pdf.txt000822638.pdf.txtExtracted Texttext/plain51703http://www.lume.ufrgs.br/bitstream/10183/225270/2/000822638.pdf.txt2d1d96ed0c76eb17d65fced32e341c4dMD52ORIGINAL000822638.pdfTexto completo (inglês)application/pdf683469http://www.lume.ufrgs.br/bitstream/10183/225270/1/000822638.pdf7c602ba5f6dd0fd22f09da4652e7576eMD5110183/2252702023-01-18 06:02:29.752532oai:www.lume.ufrgs.br:10183/225270Repositório de PublicaçõesPUBhttps://lume.ufrgs.br/oai/requestopendoar:2023-01-18T08:02:29Repositório Institucional da UFRGS - Universidade Federal do Rio Grande do Sul (UFRGS)false |
dc.title.pt_BR.fl_str_mv |
Differential macrophage activation alters the expression profile of NTPDase and Ecto-5´-nucleotidase |
title |
Differential macrophage activation alters the expression profile of NTPDase and Ecto-5´-nucleotidase |
spellingShingle |
Differential macrophage activation alters the expression profile of NTPDase and Ecto-5´-nucleotidase Zanin, Rafael Fernandes 5'-nucleotidase Ativação de macrófagos RNA mensageiro |
title_short |
Differential macrophage activation alters the expression profile of NTPDase and Ecto-5´-nucleotidase |
title_full |
Differential macrophage activation alters the expression profile of NTPDase and Ecto-5´-nucleotidase |
title_fullStr |
Differential macrophage activation alters the expression profile of NTPDase and Ecto-5´-nucleotidase |
title_full_unstemmed |
Differential macrophage activation alters the expression profile of NTPDase and Ecto-5´-nucleotidase |
title_sort |
Differential macrophage activation alters the expression profile of NTPDase and Ecto-5´-nucleotidase |
author |
Zanin, Rafael Fernandes |
author_facet |
Zanin, Rafael Fernandes Braganhol, Elizandra Bergamin, Letícia Scussel Campesato, Luis Felipe Ingrassia Zanotto Filho, Alfeu Moreira, Jose Claudio Fonseca Morrone, Fernanda Bueno Sevigny, Jean Schetinger, Maria Rosa Chitolina Wyse, Angela Terezinha de Souza Battastini, Ana Maria Oliveira |
author_role |
author |
author2 |
Braganhol, Elizandra Bergamin, Letícia Scussel Campesato, Luis Felipe Ingrassia Zanotto Filho, Alfeu Moreira, Jose Claudio Fonseca Morrone, Fernanda Bueno Sevigny, Jean Schetinger, Maria Rosa Chitolina Wyse, Angela Terezinha de Souza Battastini, Ana Maria Oliveira |
author2_role |
author author author author author author author author author author |
dc.contributor.author.fl_str_mv |
Zanin, Rafael Fernandes Braganhol, Elizandra Bergamin, Letícia Scussel Campesato, Luis Felipe Ingrassia Zanotto Filho, Alfeu Moreira, Jose Claudio Fonseca Morrone, Fernanda Bueno Sevigny, Jean Schetinger, Maria Rosa Chitolina Wyse, Angela Terezinha de Souza Battastini, Ana Maria Oliveira |
dc.subject.por.fl_str_mv |
5'-nucleotidase Ativação de macrófagos RNA mensageiro |
topic |
5'-nucleotidase Ativação de macrófagos RNA mensageiro |
description |
Macrophages are key elements in the inflammatory process, whereas depending on the micro-environmental stimulation they exhibit a pro-inflammatory (classical/M1) or an anti-inflammatory/reparatory (alternative/M2) phenotype. Extracellular ATP can act as a danger signal whereas adenosine generally serves as a negative feedback mechanism to limit inflammation. The local increase in nucleotides communication is controlled by ectonucleotidases, such as members of the ectonucleoside triphosphate diphosphohydrolase (E-NTPDase) family and ecto-59-nucleotidase/CD73 (ecto-59-NT). In the present work we evaluated the presence of these enzymes in resident mice M1 (macrophages stimulated with LPS), and M2 (macrophages stimulated with IL-4) macrophages. Macrophages were collected by a lavage of the mice (6–8 weeks) peritoneal cavity and treated for 24 h with IL-4 (10 ng/mL) or LPS (10 ng/mL). Nitrite concentrations were measured using the Greiss reaction. Supernatants were harvested to determine cytokines and the ATPase, ADPase and AMPase activities were determined by the malachite green method and HPLC analysis. The expression of selected surface proteins was evaluated by flow cytometry. The results reveal that M1 macrophages presented a decreased ATP and AMP hydrolysis in agreement with a decrease in NTPDase1, -3 and ecto-59-nucleotidase expression compared to M2. In contrast, M2 macrophages showed a higher ATP and AMP hydrolysis and increased NTPDase1, -3 and ecto-59-nucleotidase expression compared to M1 macrophages. Therefore, macrophages of the M1 phenotype lead to an accumulation of ATP while macrophages of the M2 phenotype may rapidly convert ATP to adenosine. The results also showed that P1 and P2 purinoreceptors present the same mRNA profile in both phenotypes. In addition, M2 macrophages, which have a higher ATPase activity, were less sensitive to cell death. In conclusion, these changes in ectoenzyme activities might allow macrophages to adjust the outcome of the extracellular purinergic cascade in order to fine-tune their functions during the inflammatory set. |
publishDate |
2012 |
dc.date.issued.fl_str_mv |
2012 |
dc.date.accessioned.fl_str_mv |
2021-08-06T04:41:27Z |
dc.type.driver.fl_str_mv |
Estrangeiro info:eu-repo/semantics/article |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
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article |
status_str |
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http://hdl.handle.net/10183/225270 |
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1932-6203 |
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000822638 |
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1932-6203 000822638 |
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http://hdl.handle.net/10183/225270 |
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PLoS ONE. San Francisco. Vol. 7, no. 2 (Feb. 2012), e31205, 10 f. |
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