Quantification of Monocyte Chemotactic Activity In Vivo and Characterization of Blood Monocyte Derived Macrophages

Abstract

Tissue homeostasis and repair are critically dependent on the recruitment of monocyte-derived macrophages. Both under-and over-recruitment of monocyte-derived macrophages can impair wound healing. We showed that high fat and high sugar diets promote monocyte priming and dysfunction, converting healthy blood monocytes into a hyper chemotactic phenotype poised to differentiate into macrophages with dysregulated activation profiles and impaired phenotypic plasticity. The over-recruitment of monocyte-derived macrophages and recruitment of macrophages with dysregulated activation profiles is believed to be a major contributor to the development of chronic inflammatory diseases associated with metabolic disorders, including atherosclerosis and obesity. The goal of this protocol is to quantify the chemotactic activity of blood monocytes as a biomarker for monocyte priming and dysfunction and to characterize the macrophage phenotype blood monocytes are poised to differentiate into in these mouse models. Using single cell Western blot analysis, we show that after 24 h 33%of cells recruited into MCP-1-loaded basement membrane-derived gel plugs injected into mice are monocytes and macrophages; 58% after day 3. However, on day 5, monocyte and macrophage numbers were significantly decreased. Finally, we show that this assays also allows for the isolation of live macrophages from the surgically retrieved basement membrane-derived gel plugs, which can then be subjected to subsequent characterization by single cell Western blot analysis.

Serum monocyte chemotactic protein 1 and soluble mannose receptor aid predictive diagnosis of pediatric sepsis

“…It has been reported that mechanically ventilated patients who developed ventilator-associated pneumonia had significant higher levels of MCP-1 (P<0.05) compared with patients who developed ventilator pneumonia for the purpose of developing ventilator pneumonia. Further analysis in the study revealed that when patients with ventilator pneumonia progressed to respiratory distress syndrome, the levels of MCP-1 again showed a significant elevation compared with patients who did not develop respiratory distress syndrome [17][18][19]. Several studies have pointed out that plasma MCP-1 levels can be used as a biomarker for predicting organ dysfunction, organ failure, and mortality in patients with sepsis, as well as for the diagnosis of sepsis [20,21].…”