FIG 6.

CD14 expression is largely restricted to macrophages. Expression of CD14 on liver granulocytes analyzed by flow cytometry. Single-cell suspensions from livers were prepared as described in Materials and Methods from C57BL/6 mice (n = 5) infected with S. mansoni for 12 weeks and surface stained for CD11b, F4/80, SiglecF, Gr1, and CD14 by a flow cytometer. (A) Live cells were gated for CD11b⁺ and CD11b⁻ populations. Expression of CD14 was first determined in CD11b⁺ (tinted peak) and CD11b⁻ (open peak) liver cells. (B) CD11b⁺ cells were further gated for Gr1 and F4/80 expression to separate out neutrophil (CD11b⁺Gr1⁺) and CD11b⁺F4/80⁺ populations. The CD11b⁺F4/80⁺ population was further gated for SiglecF expression with SiglecF⁺ cells as eosinophils and SiglecF-low or -negative (SiglecF⁻) cells as macrophages. Expression of CD14 was determined in neutrophil (1), eosinophil (2), and macrophage (3) populations. (C) To determine the percentages of CD14⁺ macrophages, eosinophils, or neutrophils, CD11b⁺ cells were gated for CD14 expression. CD11b⁺CD14⁺ cells were further gated for F4/80, SiglecF, and Gr1 expression (upper panel). The positive gate (control) for each population was determined by gating CD11b⁺ cells with their respective markers (Gr1, F4/80, or SiglecF) (lower panel). The same gates were then applied to the CD11b⁺CD14⁺ populations to determine the percentage of CD14-positive macrophage (CD14⁺F4/80+SiglecF⁻), eosinophil (CD14⁺SiglecF⁺), or neutrophil (CD14⁺Gr1⁺) populations (upper panel). Results represent at least 3 independent experiments.