Distinct roles of CSF family cytokines in macrophage infiltration and activation in glioma progression and injury response
Malgorzata Sielska
Laboratory of Molecular Neurobiology, Neurobiology Center, Nencki Institute of Experimental Biology, Warsaw, Poland
Search for more papers by this authorPiotr Przanowski
Laboratory of Molecular Neurobiology, Neurobiology Center, Nencki Institute of Experimental Biology, Warsaw, Poland
Search for more papers by this authorBartosz Wylot
Laboratory of Molecular Neurobiology, Neurobiology Center, Nencki Institute of Experimental Biology, Warsaw, Poland
Search for more papers by this authorKonrad Gabrusiewicz
Laboratory of Molecular Neurobiology, Neurobiology Center, Nencki Institute of Experimental Biology, Warsaw, Poland
Search for more papers by this authorMarta Maleszewska
Laboratory of Molecular Neurobiology, Neurobiology Center, Nencki Institute of Experimental Biology, Warsaw, Poland
Search for more papers by this authorMagdalena Kijewska
Laboratory of Molecular Neurobiology, Neurobiology Center, Nencki Institute of Experimental Biology, Warsaw, Poland
Search for more papers by this authorMalgorzata Zawadzka
Laboratory of Molecular Neurobiology, Neurobiology Center, Nencki Institute of Experimental Biology, Warsaw, Poland
Search for more papers by this authorJoanna Kucharska
Laboratory of Molecular Neurobiology, Neurobiology Center, Nencki Institute of Experimental Biology, Warsaw, Poland
Search for more papers by this authorKatyayni Vinnakota
Max Delbrück Center for Molecular Medicine, Cellular Neuroscience, Berlin, Germany
Search for more papers by this authorHelmut Kettenmann
Max Delbrück Center for Molecular Medicine, Cellular Neuroscience, Berlin, Germany
Search for more papers by this authorKatarzyna Kotulska
The Children's Memorial Health Institute, Warsaw, Poland
Search for more papers by this authorWieslawa Grajkowska
The Children's Memorial Health Institute, Warsaw, Poland
Search for more papers by this authorCorresponding Author
Bozena Kaminska
Laboratory of Molecular Neurobiology, Neurobiology Center, Nencki Institute of Experimental Biology, Warsaw, Poland
Correspondence to: Professor Bozena Kaminska, PhD, Laboratory of Molecular Neurobiology, Nencki Institute of Experimental Biology, 3 Pasteur Str., 02-093 Warsaw, Poland. e-mail: [email protected]Search for more papers by this authorMalgorzata Sielska
Laboratory of Molecular Neurobiology, Neurobiology Center, Nencki Institute of Experimental Biology, Warsaw, Poland
Search for more papers by this authorPiotr Przanowski
Laboratory of Molecular Neurobiology, Neurobiology Center, Nencki Institute of Experimental Biology, Warsaw, Poland
Search for more papers by this authorBartosz Wylot
Laboratory of Molecular Neurobiology, Neurobiology Center, Nencki Institute of Experimental Biology, Warsaw, Poland
Search for more papers by this authorKonrad Gabrusiewicz
Laboratory of Molecular Neurobiology, Neurobiology Center, Nencki Institute of Experimental Biology, Warsaw, Poland
Search for more papers by this authorMarta Maleszewska
Laboratory of Molecular Neurobiology, Neurobiology Center, Nencki Institute of Experimental Biology, Warsaw, Poland
Search for more papers by this authorMagdalena Kijewska
Laboratory of Molecular Neurobiology, Neurobiology Center, Nencki Institute of Experimental Biology, Warsaw, Poland
Search for more papers by this authorMalgorzata Zawadzka
Laboratory of Molecular Neurobiology, Neurobiology Center, Nencki Institute of Experimental Biology, Warsaw, Poland
Search for more papers by this authorJoanna Kucharska
Laboratory of Molecular Neurobiology, Neurobiology Center, Nencki Institute of Experimental Biology, Warsaw, Poland
Search for more papers by this authorKatyayni Vinnakota
Max Delbrück Center for Molecular Medicine, Cellular Neuroscience, Berlin, Germany
Search for more papers by this authorHelmut Kettenmann
Max Delbrück Center for Molecular Medicine, Cellular Neuroscience, Berlin, Germany
Search for more papers by this authorKatarzyna Kotulska
The Children's Memorial Health Institute, Warsaw, Poland
Search for more papers by this authorWieslawa Grajkowska
The Children's Memorial Health Institute, Warsaw, Poland
Search for more papers by this authorCorresponding Author
Bozena Kaminska
Laboratory of Molecular Neurobiology, Neurobiology Center, Nencki Institute of Experimental Biology, Warsaw, Poland
Correspondence to: Professor Bozena Kaminska, PhD, Laboratory of Molecular Neurobiology, Nencki Institute of Experimental Biology, 3 Pasteur Str., 02-093 Warsaw, Poland. e-mail: [email protected]Search for more papers by this authorNo conflicts of interest were declared.
Abstract
Gliomas attract brain-resident (microglia) and peripheral macrophages and reprogram these cells into immunosuppressive, pro-invasive cells. M-CSF (macrophage colony-stimulating factor, encoded by the CSF1 gene) has been implicated in the control of recruitment and polarization of macrophages in several cancers. We found that murine GL261 glioma cells overexpress GM-CSF (granulocyte–macrophage colony-stimulating factor encoded by the CSF2 gene) but not M-CSF when compared to normal astrocytes. Knockdown of GM-CSF in GL261 glioma cells strongly reduced microglia-dependent invasion in organotypical brain slices and growth of intracranial gliomas and extended animal survival. The number of infiltrating microglia/macrophages (Iba1+ cells) and intratumoural angiogenesis were reduced in murine gliomas depleted of GM-CSF. M1/M2 gene profiling in sorted microglia/macrophages suggests impairment of their pro-invasive activation in GM-CSF-depleted gliomas. Deficiency of M-CSF (op/op mice) did not affect glioma growth in vivo and the accumulation of Iba1+ cells, but impaired accumulation of Iba1+ cells in response to demyelination. These results suggest that distinct cytokines of the CSF family contribute to macrophage infiltration of tumours and in response to injury. The expression of CSF2 (but not CSF1) was highly up-regulated in glioblastoma patients and we found an inverse correlation between CSF2 expression and patient survival. Therefore we propose that GM-CSF triggers and drives the alternative activation of tumour-infiltrating microglia/macrophages in which these cells support tumour growth and angiogenesis and shape the immune microenvironment of gliomas. Copyright © 2013 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.
Supporting Information
Filename | Description |
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path4192-sup-0001-FigureS1.docWord document, 191.5 KB | (A) B6C3Fe a/a-Csf1op/J mice were genotyped by the TaqMan allelic discrimination method. Three populations of mice: wild-type, heterozygote, and homozygote were clearly identified. (B) Flow cytometry analysis of CD11b+CD45low microglial cells, CD11b+CD45high macrophages, and CD11b−CD45+ lymphocytes showed the diminished percentage of microglia cells in naïve op/op compared with wt mice; n = 6 mice per group, *p < 0.05 (left panel). EGFP-expressing GL261 glioma cells were intracerebrally inoculated to op/op and wild-type mice. After 15 days analysis of number of infiltrating immune cells microglia and macrophages was performed. Flow cytometry analysis of CD11b+CD45low, CD11b+CD45high cells and CD11b-CD45+ lymphocytes in tumour-bearing brains shows no significant differences in percentage of microglia in op/op and wt mice; n = 6 mice per group (right panel). |
path4192-sup-0002-FigureS2.docWord document, 205.5 KB | (A) The expression of Cd68, Itgam, Ptprc, Ccr2, and Mrc1 genes was determined using qPCR in CD11b+ cells (microglia/macrophages) isolated from sham operated and EGFP-GL261glioma-bearing brains at 15 days after implantation. (B) The levels of Cd68 mRNA were determined using qPCR in CD11b+CD45low (microglia), CD11b+CD45high (macrophages) cells isolated by FACS from EGFP-GL261 glioma-bearing brains on day 15 after implantation. Monocytes (CD11b+) were magnetically isolated from blood of EGFP-GL261 glioma-bearing mice. |
Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.
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