Abstract
Macrophage phagocytosis plays an important role in hematoma clearance after intracerebral hemorrhage (ICH). This study examined the characteristics of multinucleated giant cells (MGCs), a group of macrophages with multiple nuclei, in a mouse ICH model. Whether MGCs could be increased by treatment with a CD47 blocking antibody and decreased by treatment with clodronate liposomes were also examined. ICH was induced via autologous blood injection. Male adult C57BL/6 mice in different groups had (1) ICH alone; (2) ICH with anti-CD47 blocking antibody or control IgG; and (3) ICH with anti-CD47 antibody combined with clodronate liposomes or control liposomes. The effect of anti-CD47 antibody on MGC formation was also tested in females. Brains were harvested at days 3 or 7 for brain histology. Many MGCs were found at day 3 post-ICH, but were reduced at day 7. MGCs phagocytosed many red blood cells and were heme oxygenase-1, ferritin, YM-1, and iNOS positive. CD47 blocking antibody injection increased MGC numbers in the peri-hematomal zone and in the hematoma in both sexes. Co-injection of clodronate liposomes depleted MGCs in both the hematoma core and the peri-hematomal area. In conclusion, MGCs represent a macrophage/microglia subtype with strong phagocytosis capacity. MGCs exhibited not only an M2 but also an M1 phenotype and appeared involved in hemoglobin degradation. Anti-CD47 antibody boosted the number of MGCs, which may contribute to enhance hematoma clearance. Understanding the exact roles of MGCs in ICH may reveal novel targets for ICH treatment.
Similar content being viewed by others
References
Hankey GJ. Stroke. Lancet. 2017;389:641–54.
Xi G, Keep RF, Hoff JT. Mechanisms of brain injury after intracerebral hemorrhage. Lancet Neurol. 2006;5:53–63.
Keep RF, Andjelkovic AV, Xiang J, Stamatovic SM, Antonetti DA, Hua Y, et al. Brain endothelial cell junctions after cerebral hemorrhage: changes, mechanisms and therapeutic targets. J Cereb Blood Flow Metab. 2018;38:1255–75.
Keep RF, Hua Y, Xi G. Intracerebral haemorrhage: mechanisms of injury and therapeutic targets. Lancet Neurol. 2012;11:720–31.
Wilkinson DA, Keep RF, Hua Y, Xi G. Hematoma clearance as a therapeutic target in intracerebral hemorrhage: from macro to micro. J Cereb Blood Flow Metab. 2018;38:741–5.
Liu R, Li H, Hua Y, Keep RF, Xiao J, Xi G, et al. Early hemolysis within human intracerebral hematomas: an mri study. Transl Stroke Res. 2019;10:52–6.
Hanley DF, Thompson RE, Rosenblum M, Yenokyan G, Lane K, McBee N, et al. Efficacy and safety of minimally invasive surgery with thrombolysis in intracerebral haemorrhage evacuation (mistie iii): a randomised, controlled, open-label, blinded endpoint phase 3 trial. Lancet. 2019;393:1021–32.
Chang CF, Goods BA, Askenase MH, Hammond MD, Renfroe SC, Steinschneider AF, et al. Erythrocyte efferocytosis modulates macrophages towards recovery after intracerebral hemorrhage. J Clin Invest. 2018;128:607–24.
Jing C, Bian L, Wang M, Keep RF, Xi G, Hua Y. Enhancement of hematoma clearance with cd47 blocking antibody in experimental intracerebral hemorrhage. Stroke. 2019;50:1539–47.
Quinn MT, Schepetkin IA. Role of nadph oxidase in formation and function of multinucleated giant cells. J Innate Immun. 2009;1:509–26.
Chambers TJ, Spector WG. Inflammatory giant cells. Immunobiology. 1982;161:283–9.
Shtaya A, Bridges LR, Esiri MM, Lam-Wong J, Nicoll JAR, Boche D, et al. Rapid neuroinflammatory changes in human acute intracerebral hemorrhage. Ann Clin Transl Neurol. 2019;6:1465–79.
Yenari MA, Kauppinen TM, Swanson RA. Microglial activation in stroke: therapeutic targets. Neurotherapeutics. 2010;7:378–91.
Block ML, Zecca L, Hong JS. Microglia-mediated neurotoxicity: uncovering the molecular mechanisms. Nat Rev Neurosci. 2007;8:57–69.
Nakajima K, Yamamoto S, Kohsaka S, Kurihara T. Neuronal stimulation leading to upregulation of glutamate transporter-1 (glt-1) in rat microglia in vitro. Neurosci Lett. 2008;436:331–4.
Thored P, Heldmann U, Gomes-Leal W, Gisler R, Darsalia V, Taneera J, et al. Long-term accumulation of microglia with proneurogenic phenotype concomitant with persistent neurogenesis in adult subventricular zone after stroke. Glia. 2009;57:835–49.
Ni W, Mao S, Xi G, Keep RF, Hua Y. Role of erythrocyte cd47 in intracerebral hematoma clearance. Stroke. 2016;47:505–11.
Liu H, Hua Y, Keep RF, Xi G. Brain ceruloplasmin expression after experimental intracerebral hemorrhage and protection against iron-induced brain injury. Transl Stroke Res. 2019;10:112–9.
Ghorpade A, Persidsky Y, Swindells S, Borgmann K, Persidsky R, Holter S, et al. Neuroinflammatory responses from microglia recovered from hiv-1-infected and seronegative subjects. J Neuroimmunol. 2005;163:145–56.
Fendrick SE, Xue QS, Streit WJ. Formation of multinucleated giant cells and microglial degeneration in rats expressing a mutant cu/zn superoxide dismutase gene. J Neuroinflammation. 2007;4:9.
Vignery A. Macrophage fusion: molecular mechanisms. Methods Mol Biol. 2008;475:149–61.
Ruibal-Ares B, Riera NE, de Bracco MM. Macrophages, multinucleated giant cells, and apoptosis in hiv+ patients and normal blood donors. Clin Immunol Immunopathol. 1997;82:102–16.
McNally AK, Anderson JM. Macrophage fusion and multinucleated giant cells of inflammation. Adv Exp Med Biol. 2011;713:97–111.
McNally AK, Anderson JM. Multinucleated giant cell formation exhibits features of phagocytosis with participation of the endoplasmic reticulum. Exp Mol Pathol. 2005;79:126–35.
Normand G, King RW. Understanding cytokinesis failure. Adv Exp Med Biol. 2010;676:27–55.
Bachstetter AD, Van Eldik LJ, Schmitt FA, Neltner JH, Ighodaro ET, Webster SJ, et al. Disease-related microglia heterogeneity in the hippocampus of alzheimer’s disease, dementia with lewy bodies, and hippocampal sclerosis of aging. Acta Neuropathol Commun. 2015;3:32.
Yamada J, Jinno S. Novel objective classification of reactive microglia following hypoglossal axotomy using hierarchical cluster analysis. J Comp Neurol. 2013;521:1184–201.
Zhao H, Garton T, Keep RF, Hua Y, Xi G. Microglia/macrophage polarization after experimental intracerebral hemorrhage. Transl Stroke Res. 2015;6:407–9.
Burger P, Hilarius-Stokman P, de Korte D, van den Berg TK, van Bruggen R. Cd47 functions as a molecular switch for erythrocyte phagocytosis. Blood. 2012;119:5512–21.
Han X, Sterling H, Chen Y, Saginario C, Brown EJ, Frazier WA, et al. Cd47, a ligand for the macrophage fusion receptor, participates in macrophage multinucleation. J Biol Chem. 2000;275:37984–92.
Griesmann H, Drexel C, Milosevic N, Sipos B, Rosendahl J, Gress TM, et al. Pharmacological macrophage inhibition decreases metastasis formation in a genetic model of pancreatic cancer. Gut. 2017;66:1278–85.
Danenberg HD, Fishbein I, Gao J, Monkkonen J, Reich R, Gati I, et al. Macrophage depletion by clodronate-containing liposomes reduces neointimal formation after balloon injury in rats and rabbits. Circulation. 2002;106:599–605.
Funding
YH, RFK, and GX are supported by grants NS-091545, NS-090925, NS-096917, NS-106746, and NS-112394 from the National Institutes of Health (NIH).
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of Interest
Jialiang Wei, Ming Wang, Chaohui Jing, Richard F. Keep, Ya Hua, and Guohua Xi declare that they have no conflict of interest.
Ethical Approval
All institutional and national guidelines for the care and use of laboratory animals were followed.
Additional information
Publisher’s Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Wei, J., Wang, M., Jing, C. et al. Multinucleated Giant Cells in Experimental Intracerebral Hemorrhage. Transl. Stroke Res. 11, 1095–1102 (2020). https://doi.org/10.1007/s12975-020-00790-4
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12975-020-00790-4