Activated neutrophils exhibit enhanced phagocytosis of Cryptococcus neoformans opsonized with normal human serum

Abstract

We studied the effect of agents that activate neutrophils on phagocytosis of C. neoformans. The amount of CR3 on the surface of neutrophils was used as a marker for neutrophil activation. Surface CR3 was estimated by flow cytometry using phycoerythrin-labelled anti-CR3 (anti-Leu-15) monoclonal antibody. Phagocytosis was determined by incubation of neutrophils with cryptococci that had been preincubated with normal human serum. We found that treatment of neutrophils with (i) the chemotactic peptide N-formyl-methionyl-leucyl-phenylalanine, (ii) zymosan activated serum, (iii) supernatant fluid from a mixed leucocyte culture, or (iv) supernatant fluid from human leucocytes cultured with phytohaemagglutinin produced a dose-dependent increase in CR3 density. These agents also markedly enhanced phagocytosis of opsonized cryptococci in a parallel dose-dependent fashion. These results indicate that phagocytosis of cryptococci opsonized with normal human serum is markedly enhanced by treatment of neutrophils with reagents that stimulate neutrophils. Our results demonstrate that neutrophils activated in an appropriate manner are capable of efficient phagocytosis of encapsulated cryptococci. This potential phagocyte activity may account in part for the high natural resistance to cryptococcosis.