期刊
CYTOMETRY PART B-CLINICAL CYTOMETRY
卷 76B, 期 4, 页码 271-284出版社
WILEY
DOI: 10.1002/cyto.b.20474
关键词
monocytes; phagocytosis; subpopulations; cord blood; GFP; apoptosis; ROS
资金
- German Sepsis Society (Deutsche Sepsis Gesellschaft)
- Sanitatsrat Dr. Alexander Hubner und Gemahlin Stiftung (Deutscher Stifterverband)
Background: Neonatal sepsis is characterized by an excessive inflammatory response induced by immune cells (monocytes). We investigated the initial stage of monocyte-pathogen interaction, i.e. bacterial ingestion and degradation at the single-cell level, by comparing a new flow cytometric procedure with culture methods. We also examined the hypothesis that, in terms of phagocytosis-induced cell death (PICD), phenotype, or cytokine production, cord blood monocytes (CBMO) differ from monocytes derived from adults (peripheral blood monocytes, PBMO). Methods: Phagocytosis and intracellular degradation were assessed by means of flow cytometry and bacterial cultures of green fluorescent protein-labeled group B Streptococci (GBS) and Escherichia coli. The production of reactive oxygen species (ROS) was measured through luminol-enhanced chemiluminescence. Apoptosis, phenotype, and cytokine production were assessed through flow cytometry. Results: Flow cytometry and bacterial cultures showed no difference between phagocytosis and degradation of GBS and E coli by PBMO and CBMO. A high correlation between both methods was observed. No difference in ROS production was evident. In comparison with PBMO, CBMO apoptosis was lower after exposure to GBS and E coli. Similarities were found between nonapoptotic monocytes and pro-inflammatory monocytes. Conclusions: PICD is lower in CBMO during the early stages of monocyte-pathogen interaction. Our results emphasize that monocyte apoptosis has a potential role in tailoring the immune response in neonatal sepsis. (C) 2009 Clinical Cytometry Society
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