Figure 1

Gating strategy utilized for flow cytometric analysis of bovine peripheral blood leukocytes showing representative data from one animal. After gating on viable (propidium-iodide-negative) cells (A), cell doublets were excluded by SSC-A and SSC-H gating (B). Bovine mononuclear cells (MNC) and granulocytes (PMN) were gated based on their forward and side scatter characteristics and their percentages were calculated (C). Three-color immunofluorescence of bovine MNC with mAbs to CD172a, CD14 and CD16 defines three monocyte subsets in peripheral blood. Viable mononuclear cells, based on forward and side scatter characteristics, were gated on CD172a-positive cells (D). Dot plots of CD14 and CD16 expression display classical monocytes (CD14⁺CD16⁻, upper left), intermediate monocytes (CD14⁺CD16⁺, upper right) and nonclassical monocytes (CD14⁻CD16⁺, lower right) (E)