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Updated: May 4, 2026
Isolation and Flow Cytometric Analysis of Immune Cells from the Ischemic Mouse Brain
Published on: February 12, 2016
Allometric dose retranslation unveiled substantial immunological side effects of granulocyte colony-stimulating
Daniel-Christoph Wagner1, Claudia Pösel, Isabell Schulz
1From the Fraunhofer Institute for Cell Therapy and Immunology, Leipzig, Germany (D.-C.W., C.P., I.S., G.S., J.B., F.L., J.S., K.M., G.W.); Translational Centre for Regenerative Medicine, University of Leipzig, Leipzig, Germany (D.-C.W., J.B., G.W.); and Stroke and Neurovascular Regulation Laboratory, Massachusetts General Hospital, Harvard Medical School, Boston, MA (J.B.).
Background And Purpose:
Granulocyte colony-stimulating factor (GCSF) showed robust neuroprotective and neuroregenerative properties after stroke in rodents but failed to meet study end points in patients. Because immunologic side effects of GCSF may have escaped preclinical testing because of nonallometric dose translation, we hypothesized those as possible reasons.
Methods:
Stroke was induced in C57BL/6 mice by 45-minute filament middle cerebral artery occlusion. GCSF was administered at 50 and 832.5 μg/kg body weight. Treatment was controlled by vehicle injection, sham surgery, and naive animals. Immune cell counts were assessed in blood, spleen, and brain by multidimensional flow cytometry 1 day after stroke.
Results:
High-dose GCSF significantly altered myeloid and T-cell subpopulations in blood and spleen and caused a tremendous increase of monocytes/macrophages infiltrating the ischemic brain.
Conclusions:
Dose-dependent immunomodulation superimposes central nervous system-specific effects of GCSF after stroke. Adaption of dose or treatment time may overcome this drawback.
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