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

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

Autor Gomes, Natalia Estevam Autor UNIFESP Google Scholar
Brunialti, Milena Karina Coló Autor UNIFESP Google Scholar
Mendes, Marialice Erdelyi Autor UNIFESP Google Scholar
Freudenberg, M. Google Scholar
Galanos, C. Google Scholar
Salomão, Reinaldo Autor UNIFESP Google Scholar
Instituição Universidade Federal de São Paulo (UNIFESP)
Max-Planck-Institute for Immunobiology
Resumo 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, doubling after 5 min on monocytes, while down-regulation of CXCR2 was observed on neutrophils, reaching a 50% reduction after 6 h. LPS induced low production of ROS and NO. This study shows a complex LPS-induced cell surface receptor modulation on human monocytes and neutrophils, with up- and down-regulation depending on the receptor. R- and S-form LPS activate human neutrophils similarly, despite the low CD14 expression, if the stimulation occurs in whole blood.
Palavra-chave Lipopolysaccharide
Neutrophils
Monocytes
Cell surface receptors
Reactive oxygen species
Nitric oxide
Idioma Inglês
Financiador Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
Número do financiamento FAPESP: 2006/58744-1
Data de publicação 2010-09-01
Publicado em Brazilian Journal of Medical and Biological Research. Associação Brasileira de Divulgação Científica, v. 43, n. 9, p. 853-858, 2010.
ISSN 0100-879X (Sherpa/Romeo, fator de impacto)
Publicador Associação Brasileira de Divulgação Científica
Extensão 853-858
Fonte http://dx.doi.org/10.1590/S0100-879X2010007500078
Direito de acesso Acesso aberto Open Access
Tipo Artigo
Web of Science WOS:000283262800007
SciELO S0100-879X2010000900007 (estatísticas na SciELO)
Endereço permanente http://repositorio.unifesp.br/handle/11600/5910

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