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Blood, 1 May 2006, Vol. 107, No. 9, pp. 3520-3526.
Prepublished online as a Blood First Edition Paper on January 3, 2006; DOI 10.1182/blood-2005-10-4285.
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Submitted October 31, 2005
Accepted December 28, 2005
Monocyte-derived CXCL7 peptides in the marrow microenvironment (ME)
Manoj M Pillai, Mineo Iwata, Norihiro Awaya, Lynn Graf, and Beverly Torok-Storb*
Clinical Research Division, Fred Hutchinson Cancer Research Center, Seattle, WA, USA; Department of Medicine, University of Washington School of Medicine, Seattle, WA, USA
Clinical Research Division, Fred Hutchinson Cancer Research Center, Seattle, WA, USA
* Corresponding author; email: btorokst{at}fhcrc.org; blarson@fhcrc.org.
The marrow microenvironment consists of several different interacting cell types including hematopoietic-derived monocyte/macrophages and nonhematopoietic-derived stromal cells. Gene expression profiles of stromal cells and monocytes cultured together differ from those of each population alone. Here we report that CXCL7 gene expression, previously described as limited to the megakaryocyte lineage, is expressed by monocytes co-cultured with stromal cells. CXCL7 gene expression was confirmed by quantitative RT-PCR, and secretion of protein was detected by ELISA and Western blot. At least two stromal-derived activities, one yet to be identified, were required for optimal expression of CXCL7 by monocytes. NAP-2, the shortest form of CXCL7 detected in the co-culture media, was confirmed to decrease the size and number of CFU-Meg colonies. The pro-peptide LDGF, previously reported to be mitogenic for fibroblasts, was not secreted by stimulated monocytes. The recombinant form of LDGF produced in a prokaryotic expression system did not have biological activity in our hands. The monocytic source of CXCL7 was also detected by immunohistochemistry in normal bone marrow biopsies, indicating an in vivo function. We conclude that stromal-stimulated monocytes can serve as an additional source for CXCL7 peptides in the microenvironment and may contribute to the local regulation of megakaryocytopoiesis.
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