Journal of Leukocyte Biology Myeloid cells, immune suppression, tumor immunology
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Published online before print December 30, 2005
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© by The Society for Leukocyte Biology
Journal of Leukocyte Biology, doi:10.1189/jlb.0505248

Received for publication May 9, 2005.
Revised November 4, 2005.
Accepted for publication November 7, 2005.

Article

Localization of human neutrophil interleukin-8 (CXCL-8) to organelle(s) distinct from the classical granules and secretory vesicles

Sara Pellmé *@, Matthias Mörgelin {dagger}, Hans Tapper {dagger}, Ulf-Henrik Mellqvist {ddagger}, Claes Dahlgren *, and Anna Karlsson *

*Department of Rheumatology and Inflammation Research, University of Göteborg, Sweden; {dagger}Department of Cell and Molecular Biology, Lund University, Sweden; and {ddagger}Hematology Section, Sahlgrenska University Hospital, Göteborg, Sweden

@ To whom correspondence should be addressed. E-mail: sara.pellme{at}microbio.gu.se.


  Abstract

Mature human neutrophils contain small amounts of interleukin-8 [CXC chemokine ligand 8 (CXCL-8)], which upon proinflammatory activation, increases significantly. It has been suggested that the CXCL-8 content of resting human neutrophils is stored in the secretory vesicles. Here, we have used a fractionation technique, which allows isolation of these vesicles, and we find that CXCL-8 neither colocalizes with the secretory vesicles nor with markers of any of the classical neutrophil granules. To increase resolution in the system, we induced CXCL-8 production by lipopolysaccharide. After 8 h of stimulation, CXCL-8 was visualized within the cell using immunoelectron microscopy. The images revealed CXCL-8-containing stuctures resembling neutrophil granules, and these were distinct from all known neutrophil organelles, as shown by double immunostaining. Further, the CXCL-8 organelle was present in nonstimulated neutrophil cytoplasts, entities lacking all other known granules and secretory vesicles. Upon fractionation of the cytoplasts, CXCL-8 was found to partly cofractionate with calnexin, a marker for endoplasmic reticulum (ER). Thus, part of CXCL-8 may be localized to the ER or ER-like structures in the neutrophil.

Key Words: chemokines • inflammation • subcellular organization • cell activation




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