Annexin III translocates to the periphagosomal region when neutrophils ingest opsonized yeast

After phagocytosis, killing and digestion of ingested microorganisms depend on fusion of phagocytic vesicle membranes with membranes of intracellular vesicles (azurophil and specific granules). There is considerable evidence that phagosome-granule membrane fusion is regulated by transient increases in intracellular ionized Ca2+. In previous studies, we found that a cytosolic Ca2(+)-dependent membrane-binding protein, annexin III, represents over 1% of the total protein of human neutrophils and promotes tight contact between membranes of isolated specific granules in vitro. To determine whether annexin III localizes to the region of phagosome-granule membrane fusion in vivo, we used a monospecific polyclonal antibody to stain fixed, permeabilized neutrophils that had ingested opsonized yeast. We found that annexin III concentrates in the region surrounding the phagosome. Annexin III was concentrated ninefold in the periphagosomal region compared with the cell body, as demonstrated by laser scanning confocal microscopy. Periphagosomal translocation of annexin III occurred whether yeast were opsonized with IgG, complement, or both, and persisted for at least 1 h after phagocytosis. This is not a general phenomenon, inasmuch as calmodulin was as abundant in the cell body as in the periphagosomal region. These findings imply that annexin III plays a specialized role in the metabolic and structural events that accompany phagocytosis.