Localization of eosinophil-derived neurotoxin and eosinophil cationic protein in neutrophilic leukocytes

Eosinophil-derived neurotoxin (EDN) and eosinophil cationic protein (ECP) are generally regarded as eosinophil-specific proteins. We tested whether EDN and ECP are present iu mature neutrophils, By indirect immunofluorescence, both eosinophils and neutrophils stained with antibodies to EDN and ECP. Lq-sates of purified (<0.1% eosinophil contamination) neutrophils contained EDN, 112 +/- 4 ng/10(6) cells, and ECP, 163 +/- 2 ng/10(6) cells, whereas eosinophil major basic protein (MBP) was not detectable. Electron microscopic examination of immunogold-labeled buffy coat cells stained with EDN antibody showed that EDN is localized to neutrophil granules. Finally, EDN mRNA was detected in lysates of highly purified neutrophils (0.001% eosinophil contamination) by the reverse transcription-polymerase chain reaction, We conclude that proteins that are either identical to or immunologically crossreactive with EDN and ECP are present in neutrophils and that EDN is synthesized and localized to neutrophil granules. Thus, caution must be exercised in interpreting the presence of EDN and ECP as specific markers of eosinophil-associated inflammation in human disease.