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Evidence for small intracellular vesicles in human blood phagocytes
containing cytochrome b558 and the adhesion molecule CD11b/CD18
J Calafat, TW Kuijpers, H Janssen, N Borregaard, AJ Verhoeven and D Roos
Division of Cell Biology, The Netherlands Cancer Institute, Amsterdam.
Human neutrophils contain a rapidly mobilizable pool of so-called secretory
vesicles distinct from the azurophil granules and specific granules. Using
human albumin as a marker for these intracellular vesicles in
immuno-electron microscopy, we found that part of the cytochrome b558 in
non-purified whole blood neutrophils colocalized in these vesicles. This
was detected with monoclonal antibody (MoAb) CLB- 48, binding to the high
molecular weight subunit of cytochrome b558. Approximately 65% of the
albumin-containing vesicles showed MoAb CLB-48 labeling. This was also
found in eosinophilic granulocytes and in monocytes. Cytofluorimetric
determination of cytochrome b558 expression on the plasma membrane of
intact, nonpurified granulocytes (and monocytes) with MoAb 7D5, which is
directed against an extracellular epitope of cytochrome b558, did not show
any binding. However, granulocytes (and monocytes) significantly bound 7D5
after density centrifugation. The positive binding of 7D5 to purified
neutrophilic granulocytes correlated with a strongly reduced labeling of
cytochrome b558 in the albumin-positive vesicles. Binding of CD11b MoAb
CLB-B2.12 to the alpha subunit of the complement receptor type 3 (CR3) on
the surface of intact, nonpurified neutrophils was detected to a limited
extent in whole blood samples, but was strongly increased upon density
gradient centrifugation of the cells, as we have described before.
Investigation at the ultrastructural level showed that the CD11b antigen
codistributed with albumin in vesicular structures in nonpurified
phagocytes, especially in neutrophils and eosinophils. Together, these data
substantiate the idea of an intracellular store that can be easily
mobilized (even under the simple stress condition of density gradient
centrifugation). Such mobilization may result in the expression of
cytochrome b558 on the plasma membrane, as was indicated in this study.
Apart from cytochrome b558, several other surface membrane molecules, as we
show here for the integrin CD11b/CD18 (CR3), are probably also located in
these rapidly mobilizable intracellular vesicles.
Volume 81,
Issue 11,
pp. 3122-3129,
06/01/1993
Copyright © 1993 by The American Society of Hematology
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