Figure 10. LPC treatment of primary human PMNs increases primary granule fusion with Gc phagosomes.

(A–B) PMNs were infected with Gc for 45 min, then given media with or without LPC. Intracellular and extracellular Gc were discriminated from one another along with an antibody raised against neutrophil elastase (NE). Extracellular Gc appear red/blue, intracellular Gc appear blue only, and neutrophil elastase staining appears green. In (A) arrowheads indicate bacterial phagosomes positive for granule proteins, while arrows indicate phagosomes negative for granule proteins. The percent of phagosomes exhibiting enrichment of neutrophil elastase with and without LPC treatment is reported in B. (C–E) PMNs were infected with Gc and left untreated (C) or treated with LPC (D). Viable Gc (green) and nonviable Gc (red) were discriminated using Baclight viability dyes SYTO9 and propidium iodide, and extracellular Gc were labeled with soybean lectin (blue). Extracellular viable Gc appear teal, intracellular nonviable Gc appear red, and intracellular viable Gc appear green. In C and D arrows indicate viable, intracellular Gc and arrow heads indicate nonviable, intracellular Gc. The percent of viable extracellular and intracellular Gc in untreated and LPC-treated PMNs is reported in E. Asterisks indicate P < 0.05 by Student’s two-tailed t test.