Highly purified human peripheral blood monocytes produce IL-6 but not TNFα in response to angiotensin II

Abstract

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 AT1R 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-α 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-α mRNA transcripts or protein secretion. Conclusions: We provide evidence that a CD14+/CD16- subset of highly purified human monocytes express AT1R and respond to AngII exposure in vitro by producing IL-6 but not TNF-α.