Journal
JOURNAL OF THE RENIN-ANGIOTENSIN-ALDOSTERONE SYSTEM
Volume 12, Issue 3, Pages 295-303Publisher
SAGE PUBLICATIONS LTD
DOI: 10.1177/1470320310391332
Keywords
Monocytes; TNF-alpha; IL-6; Inflammation; Angiotensin II; AT(1)R
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Funding
- Nova Scotia Health Research Foundation [42311]
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Hypothesis: Monocytes produce pro-inflammatory cytokines in response to Angiotensin II (AngII). Introduction: AngII has been suggested by many to be pro-inflammatory and likely to contribute to the migration of leukocytes in patients with cardiovascular conditions. Materials and methods: Monocytes were purified from peripheral blood mononuclear cells (PBMCs) by negative selection using antibodies conjugated to magnetic beads. Detection of CD14(+) and AT(1)R expression was achieved by double-labeling flow cytometry. Highly purified monocytes were then stimulated with AngII (6 and 24 h) to assess IL-6 and TNF-alpha transcript levels by qRT-PCR and protein secretion by ELISA. Results: Monocytes comprised 9.7 +/- 2.0% of the PBMCs. Monocyte isolation by negative selection yielded a purity of up to 99.8%. We demonstrated AT1R expression on 9.5 +/- 0.3% of highly purifed CD14(+)/CD16(-) monocytes. Stimulation of highly purified monocytes with AngII resulted in increased transcript levels of IL-6 at 6 h but not at 24 h, and increased secretion of IL-6 in a dose-dependent manner compared with controls (p < 0.01). Conversely, there was no increase in TNF-alpha mRNA transcripts or protein secretion. Conclusions: We provide evidence that a CD14(+)/CD16(-) subset of highly purified human monocytes express AT(1)R and respond to AngII exposure in vitro by producing IL-6 but not TNF-alpha.
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