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Assessing the delivery of neutrophils to tissues in neutropenia
DG Wright, AI Meierovics and JM Foxley
Studies of neutrophil kinetics in neutropenic individuals, as well as
clinical observations of variability in the occurrence of infection among
patients with neutropenia, have suggested that blood neutrophil counts may
not uniformly reflect the effective delivery of neutrophils to
extravascular tissues where the cells perform their principal host defense
functions. To evaluate this possibility we developed a sensitive,
reproducible method of measuring the extravascular delivery of neutrophils
to a normal mucosal site of neutrophil turnover. This method is based upon
the quantification of neutrophils recoverable from saline mouth wash
specimens. Twenty-five mL specimens, obtained in a controlled manner from
neutropenic patients and normal subjects, were centrifuged and the
sediments resuspended in 1.0 mL Hank's buffer with 2 micrograms acridine
orange, incubated at 37 degrees C for 15 minutes, and then examined in a
hemocytometer chamber by fluorescence microscopy. Neutrophils could be
clearly distinguished by their characteristic fluorescence and were
counted. With this method as few as 1,500 neutrophils were detected
reliably in mouth wash specimens. Mucosal neutrophil counts varied less
than 10% with repeated sampling of individual subjects over 5-day periods
and were consistently greater than 1.3 X 10(5)/specimen in non-neutropenic
individuals. Although profound neutropenia was generally reflected by lower
than normal oral mucosal neutrophil counts, these counts were significantly
higher in individuals with chronic severe neutropenia (blood neutrophils
less than 300/mm3) than in patients with acute neutropenia of comparable
severity that had developed following chemotherapy. Also, in individuals
recovering from profound neutropenia, neutrophils usually reappeared
earlier in mouth wash specimens than in blood, and oral mucosal neutrophil
counts attained recovery levels more rapidly than did blood counts. This
phenomenon was particularly evident in an individual with cyclic
neutropenia. Moreover, mucosal neutrophils could occasionally be detected
in profoundly neutropenic patients when neutrophils were not present in
blood samples. These findings indicate that mucosal neutrophil counts in
individuals with neutropenia provide information about the delivery of
neutrophils to tissues that may not be apparent from blood neutrophil
counts alone.
Volume 67,
Issue 4,
pp. 1023-1030,
04/01/1986
Copyright © 1986 by The American Society of Hematology
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