Translational Stroke Research

, Volume 3, Supplement 1, pp 125–131

Gr1+ Macrophages and Dendritic Cells Dominate the Inflammatory Infiltrate 12 h After Experimental Intracerebral Hemorrhage

  • Matthew D. Hammond
  • Youxi Ai
  • Lauren H. Sansing
Original Article


Intracerebral hemorrhage (ICH) is a devastating disease lacking an effective treatment. While the initial injury occurs within minutes, an inflammatory response contributes to ongoing tissue damage over hours to days. Relatively little is known about leukocyte trafficking into the brain in the hours after ICH onset. Understanding these events may lead to identification of new therapeutic targets. Using the blood injection mouse model of ICH, the numbers of leukocytes in the ipsilateral and contralateral brain were quantified by flow cytometry 12 h after surgery. Perihematomal inflammation was confirmed by histology and chemokines and cytokines in the brain quantified by multiplex ELISA. Few neutrophils were detected in the brain 12 h after ICH. The majority of leukocytes consisted of inflammatory macrophages (CD45.1hiCD3Ly6GCD11cCD11b+Gr1+ cells) and inflammatory dendritic cells (CD45.1hiCD3Ly6GCD11cintCD11b+Gr1+ cells). Microglia numbers did not differ between the hemispheres. These results indicate that blood-derived monocyte populations traffic into brain early after ICH and outnumber neutrophils at 12 h.


Intracerebral hemorrhage Inflammation Stroke Macrophages Dendritic cells Neutrophils Neuroinflammation 


  1. 1.
    Qureshi AI, Mendelow AD, Hanley DF. Intracerebral haemorrhage. Lancet. 2009;373:1632–44.PubMedCrossRefGoogle Scholar
  2. 2.
    Wang J, Dore S. Inflammation after intracerebral hemorrhage. J Cereb Blood Flow Metab. 2007;27:894–908.PubMedCrossRefGoogle Scholar
  3. 3.
    Kane PJ, Modha P, Strachan RD, Cook S, Chambers IR, Clayton CB, et al. The effect of immunosuppression on the development of cerebral oedema in an experimental model of intracerebral haemorrhage: whole body and regional irradiation. J Neurol Neurosurg Psychiatry. 1992;55:781–6.PubMedCrossRefGoogle Scholar
  4. 4.
    Lee ST, Chu K, Jung KH, Kim SJ, Kim DH, Kang KM, et al. Anti-inflammatory mechanism of intravascular neural stem cell transplantation in haemorrhagic stroke. Brain. 2008;131:616–29.PubMedCrossRefGoogle Scholar
  5. 5.
    Sansing LH, Kasner SE, McCullough L, Agarwal P, Welsh FA, K. K. Autologous blood injection to model spontaneous intracerebral hemorrhage in mice. Journal of Visualized Experiments 2011; Accessed 9 Feb 2012.
  6. 6.
    Del Bigio MR, Yan H-J, Buist R, Peeling J, del Zoppo GJ. Experimental intracerebral hemorrhage in rats: magnetic resonance imaging and histopathological correlates. Stroke. 1996;27:2312–20.PubMedCrossRefGoogle Scholar
  7. 7.
    Xue M, Del Bigio MR. Intracerebral injection of autologous whole blood in rats: time course of inflammation and cell death. Neurosci Lett. 2000;283:230–2.PubMedCrossRefGoogle Scholar
  8. 8.
    Gong C, Hoff JT, Keep RF. Acute inflammatory reaction following experimental intracerebral hemorrhage in rat. Brain Res. 2000;871:57–65.PubMedCrossRefGoogle Scholar
  9. 9.
    Auffray C, Sieweke MH, Geissmann F. Blood monocytes: development, heterogeneity, and relationship with dendritic cells. Annu Rev Immunol. 2009;27:669–92.PubMedCrossRefGoogle Scholar
  10. 10.
    Serbina NV, Salazar-Mather TP, Biron CA, Kuziel WA, Pamer EG. Tnf/inos-producing dendritic cells mediate innate immune defense against bacterial infection. Immunity. 2003;19:59–70.PubMedCrossRefGoogle Scholar
  11. 11.
    King IL, Dickendesher TL, Segal BM. Circulating ly-6c + myeloid precursors migrate to the cns and play a pathogenic role during autoimmune demyelinating disease. Blood. 2009;113:3190–7.PubMedCrossRefGoogle Scholar
  12. 12.
    Dimitrijevic OB, Stamatovic SM, Keep RF, Andjelkovic AV. Absence of the chemokine receptor ccr2 protects against cerebral ischemia/reperfusion injury in mice. Stroke. 2007;38:1345–53.PubMedCrossRefGoogle Scholar
  13. 13.
    Schilling M, Strecker JK, Ringelstein EB, Schabitz WR, Kiefer R. The role of cc chemokine receptor 2 on microglia activation and blood-borne cell recruitment after transient focal cerebral ischemia in mice. Brain Res. 2009;1289:79–84.PubMedCrossRefGoogle Scholar
  14. 14.
    Yao Y, Tsirka SE. The CCL2-CCR2 system affects the progression and clearance of intracerebral hemorrhage. Glia. 2012;60(6):908–18.PubMedCrossRefGoogle Scholar
  15. 15.
    Auffray C, Fogg D, Garfa M, Elain G, Join-Lambert O, Kayal S, et al. Monitoring of blood vessels and tissues by a population of monocytes with patrolling behavior. Science. 2007;317:666–70.PubMedCrossRefGoogle Scholar
  16. 16.
    Nahrendorf M, Swirski FK, Aikawa E, Stangenberg L, Wurdinger T, Figueiredo JL, et al. The healing myocardium sequentially mobilizes two monocyte subsets with divergent and complementary functions. J Exp Med. 2007;204:3037–47.PubMedCrossRefGoogle Scholar
  17. 17.
    Martinez FO, Gordon S, Locati M, Mantovani A. Transcriptional profiling of the human monocyte-to-macrophage differentiation and polarization: new molecules and patterns of gene expression. J Immunol. 2006;177:7303–11.PubMedGoogle Scholar
  18. 18.
    Nakano H, Lin KL, Yanagita M, Charbonneau C, Cook DN, Kakiuchi T, et al. Blood-derived inflammatory dendritic cells in lymph nodes stimulate acute t helper type 1 immune responses. Nat Immunol. 2009;10:394–402.PubMedCrossRefGoogle Scholar
  19. 19.
    Anzai A, Anzai T, Nagai S, Maekawa Y, Naito K, Kaneko H, et al. Regulatory role of dendritic cells in post-infarction healing and left ventricular remodeling. Circulation. 2012;125(10):1234–45.PubMedCrossRefGoogle Scholar
  20. 20.
    Sansing LH, Harris TH, Welsh FA, Kasner SE, Hunter CA, Kariko K. Toll-like receptor 4 contributes to poor outcome after intracerebral hemorrhage. Ann Neurol. 2011;70:646–56.PubMedCrossRefGoogle Scholar
  21. 21.
    Sagar D, Foss C, El Baz R, Pomper MG, Khan ZK, Jain P. Mechanisms of dendritic cell trafficking across the blood–brain barrier. J Neuroimmune Pharmacol. 2012;7(1):74–94.PubMedCrossRefGoogle Scholar
  22. 22.
    Xi G, Hua Y, Keep RF, Younger JG, Hoff JT. Systemic complement depletion diminishes perihematomal brain edema in rats. Stroke. 2001;32:162–7.PubMedCrossRefGoogle Scholar
  23. 23.
    Hua Y, Wu J, Keep RF, Nakamura T, Hoff JT, Xi G. Tumor necrosis factor-alpha increases in the brain after intracerebral hemorrhage and thrombin stimulation. Neurosurgery. 2006;58:542–50. discussion 542–550.PubMedGoogle Scholar
  24. 24.
    Mayne M, Ni W, Yan HJ, Xue M, Johnston JB, Del Bigio MR, et al. Antisense oligodeoxynucleotide inhibition of tumor necrosis factor-alpha expression is neuroprotective after intracerebral hemorrhage. Stroke. 2001;32:240–8.PubMedCrossRefGoogle Scholar
  25. 25.
    Liesz A, Middelhoff M, Zhou W, Karcher S, Illanes S, Veltkamp R. Comparison of humoral neuroinflammation and adhesion molecule expression in two models of experimental intracerebral hemorrhage. Exp Transl Stroke Med. 2011;3:11.PubMedCrossRefGoogle Scholar
  26. 26.
    Carmichael ST, Vespa PM, Saver JL, Coppola G, Geschwind DH, Starkman S, et al. Genomic profiles of damage and protection in human intracerebral hemorrhage. J Cereb Blood Flow Metab. 2008;28:1860–75.PubMedCrossRefGoogle Scholar
  27. 27.
    Guillemin GJ, Brew BJ. Microglia, macrophages, perivascular macrophages, and pericytes: a review of function and identification. J Leukoc Biol. 2004;75:388–97.PubMedCrossRefGoogle Scholar
  28. 28.
    Renno T, Krakowski M, Piccirillo C, Lin JY, Owens T. Tnf-alpha expression by resident microglia and infiltrating leukocytes in the central nervous system of mice with experimental allergic encephalomyelitis. Regulation by th1 cytokines. J Immunol. 1995;154:944–53.PubMedGoogle Scholar
  29. 29.
    Jin R, Yang G, Li G. Inflammatory mechanisms in ischemic stroke: role of inflammatory cells. J Leukoc Biol. 2010;87:779–89.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Matthew D. Hammond
    • 1
  • Youxi Ai
    • 1
  • Lauren H. Sansing
    • 1
    • 2
    • 3
  1. 1.Department of NeuroscienceUniversity of Connecticut Health CenterFarmingtonUSA
  2. 2.Department of NeurologyUniversity of Connecticut Health CenterFarmingtonUSA
  3. 3.The Stroke Center at Hartford HospitalHartfordUSA

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