Toll-like receptor 2 deficiency leads to delayed exacerbation of ischemic injury

Journal of Neuroinflammation

Jasna Kriz¹  Srecko Gajovic³  Yuan-Cheng Weng²  Dunja Gorup³  Mélanie Lalancette–Hébert¹  Pierre Cordeau¹  Ivan Bohacek² 

[1] Department of Psychiatry and Neuroscience, Faculty of Medicine, Université Laval, Québec, Canada;Centre de Recherche du Centre Hospitalier de l’Université Laval CHUL (CHUQ), 2705 boulevard Laurier, Québec, QC, G1V 4G2, Canada;Laboratory for Neurogenetics and Developmental Genetics, Croatian Institute for Brain Research, School of Medicine, University of Zagreb, Salata 12, Zagreb, HR-10000, Croatia

关键词: TLR2−/− mice;    stroke;    neuroinflammation;    microglia/macrophages;    IGF-1;    Apoptosis;   

Others  :  1160322
DOI  :  10.1186/1742-2094-9-191

 received in 2012-01-12, accepted in 2012-07-13,  发布年份 2012

【 摘 要 】

Background

Using a live imaging approach, we have previously shown that microglia activation after stroke is characterized by marked and long-term induction of the Toll-like receptor (TLR) 2 biophotonic signals. However, the role of TLR2 (and potentially other TLRs) beyond the acute innate immune response and as early neuroprotection against ischemic injury is not well understood.

Methods

TLR2−/− mice were subjected to transient middle cerebral artery occlusion followed by different reperfusion times. Analyses assessing microglial activation profile/innate immune response were performed using in situ hybridization, immunohistochemistry analysis, flow cytometry and inflammatory cytokine array. The effects of the TLR2 deficiency on the evolution of ischemic brain injury were analyzed using a cresyl violet staining of brain sections with appropriate lesion size estimation.

Results

Here we report that TLR2 deficiency markedly affects post-stroke immune response resulting in delayed exacerbation of the ischemic injury. The temporal analysis of the microglia/macrophage activation profiles in TLR2−/− mice and age-matched controls revealed reduced microglia/macrophage activation after stroke, reduced capacity of resident microglia to proliferate as well as decreased levels of monocyte chemotactic protein-1 (MCP-1) and consequently lower levels of CD45^high/CD11b⁺ expressing cells as shown by flow cytometry analysis. Importantly, although acute ischemic lesions (24 to 72 h) were smaller in TLR2−/− mice, the observed alterations in innate immune response were more pronounced at later time points (at day 7) after initial stroke, which finally resulted in delayed exacerbation of ischemic lesion leading to larger chronic infarctions as compared with wild-type mice. Moreover, our results revealed that TLR2 deficiency is associated with significant decrease in the levels of neurotrophic/anti-apoptotic factor Insulin-like growth factor-1 (IGF-1), expressed by microglia in the areas both in and around ischemic lesion.

Conclusion

Our results clearly suggest that optimal and timely microglial activation/innate immune response is needed to limit neuronal damage after stroke.

【 授权许可】

   
2012 Bohacek et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

Figure 5.

Figure 6.

Figure 7.

【 参考文献 】

• [1]Lo EH, Dalkara T, Moskowitz MA: Mechanisms, challenges and opportunities in stroke. Nat Rev Neurosci 2003, 4:399-415.
• [2]Kempermann G, Neumann H: Neuroscience. Microglia: the enemy within? Science 2003, 302:1689-1690.
• [3]Lalancette-Hebert M, Gowing G, Simard A, Weng YC, Kriz J: Selective ablation of proliferating microglial cells exacerbates ischemic injury in the brain. J Neurosci 2007, 27:2596-2605.
• [4]Kriz J, Lalancette-Hebert M: Inflammation, plasticity and real-time imaging after cerebral ischemia. Acta Neuropathol 2009, 117:497-509.
• [5]Iadecola C, Anrather J: Stroke research at a crossroad: asking the brain for directions. Nat Neurosci 2011, 14:1363-1368.
• [6]Akira S, Takeda K: Toll-like receptor signaling. Nat Rev Immunol 2004, 4:499-511.
• [7]Cao CX, Yang QW, Lv FL, Cui J, Fu HB, Wang JZ: Reduced cerebral ischemia-reperfusion injury in Toll-like receptor 4 deficient mice. Biochem Biophys Res Commun 2007, 353:509-514.
• [8]Caso JR, Pradillo JM, Hurtado O, Lorenzo P, Moro MA, Lizasoain I: Toll-like receptor 4 is involved in brain damage and inflammation after experimental stroke. Circulation 2007, 115:1599-1608.
• [9]Ziegler G, Harhausen D, Schepers C, Hoffmann O, Röhr C, Prinz V, König J, Lehrach H, Nietfeld W, Trendelenburg G: TLR2 has a detrimental role in mouse transient focal cerebral ischemia. Biochem Biophys Res Commun 2007, 359:574-579.
• [10]Ziegler G, Freyer D, Harhausen D, Khojasteh U, Nietfeld W, Trendelenburg G: Blocking TLR2 in vivo protects against accumulation of inflammatory cells and neuronal injury in experimental stroke. J Cereb Blood Flow Metab 2010, 31:757-766.
• [11]Lalancette-Hébert M, Phaneuf D, Soucy G, Weng YC, Kriz J: Live imaging of Toll-like receptor 2 response in cerebral ischaemia reveals a role of olfactory bulb microglia as modulators of inflammation. Brain 2009, 132:940-954.
• [12]Arumugam TV, Okun E, Tang SC, Thundyil J, Taylor SM, Woodruff TM: Toll-like receptors in ischemia-reperfusion injury. Shock 2009, 32:4-16.
• [13]Gordon S: Pattern recognition receptors: doubling up for the innate immune response. Cell 2002, 111:927-930.
• [14]Janeway CA, Medzhitov R: Innate immune recognition. Annu Rev Immunol 2002, 20:197-216.
• [15]Medzhitov R, Preston-Hurlburt P, Janeway CA: A human homologue of the Drosophila Toll protein signals activation of adaptive immunity. Nature 1997, 388:394-397.
• [16]Hanisch UK, Johnson TV, Kipnis J: Toll-like receptors: roles in neuroprotection? Trends Neurosci 2008, 31:176-182.
• [17]Asea A, Rehli M, Kabingu E, Boch JA, Bare O, Auron PE, Stevenson MA, Calderwood SK: Novel signal transduction pathway utilized by extracellular HSP70: role of toll-like receptor (TLR) 2 and TLR4. J Biol Chem 2002, 277:15028-15034.
• [18]Kariko K, Weissman D, Welsh FA: Inhibition of Toll-like receptor and cytokine signaling – a unifying theme in ischemic tolerance. J Cereb Blood Flow Metab 2004, 24:1288-1304.
• [19]Matzinger P: The danger model: a renewed sense of self. Science 2002, 296:301-305.
• [20]Lehnardt S, Lehrmann S, Kaul D, Tschimmel K, Hoffmann O, Cho S, Krueger C, Nitsch R, Meisel A, Weber JR: Toll-like receptor 2 mediates CNS injury in focal cerebral ischemia. J Neuroim 2007, 190:28-33.
• [21]Tang SC, Arumugam TV, Xu X, Cheng A, Mughal MR, Jo DG, Lathia JD, Siler DA, Chigurupati S, Ouyang X, Magnus T, Camandola S, Mattson MP: Pivotal role for neuronal Toll-like receptors in ischemic brain injury and functional deficits. Proc Natl Acad Sci U S A 2007, 104:13798-13803.
• [22]Aliprantis AO, Yang RB, Weiss DS, Godowski P, Zychlinsky A: The apoptotic signaling pathway activated by Toll-like receptor-2. EMBO J 2000, 19:3325-3336.
• [23]Babcock AA, Wirenfeldt M, Holm T, Nielsen HH, Dissing-Olesen L, Toft-Hansen H, Millward JM, Landmann R, Rivest S, Finsen B, Owens T: Toll-like receptor 2 signaling in response to brain injury: an innate bridge to neuroinflammation. J Neurosci 2006, 26:12826-12837.
• [24]Hua F, Ma J, Ha T, Kelley JL, Kao RL, Schweitzer JB, Kalbfleisch JH, Williams DL, Li C: Differential roles of TLR2 and TLR4 in acute focal cerebral ischemia/reperfusion injury in mice. Brain Res 2009, 1262:100-108.
• [25]Beaulieu JM, Kriz J, Julien JP: Induction of peripherin expression in subsets of brain neurons after lesion injury or cerebral ischemia. Brain Res 2002, 946:153-161.
• [26]Cordeau P, Lalancette-Hebert M, Weng YC, Kriz J: Live imaging of neuroinflammation reveals sex and estrogen effects on astrocyte response to ischemic injury. Stroke 2008, 39:935-942.
• [27]Campanella M, Sciorati C, Tarozzo G, Beltramo M: Flow cytometric analysis of inflammatory cells in ischemic rat brain. Stroke 2002, 33:586-592.
• [28]Lalancette–Hébert M, Swarup V, Beaulieu M, Bohacek I, Abdelhamid E, Weng Y-C, Sato S, Kriz J: Galectin-3 is required for resident microglia activation and proliferation in response to ischemic injury. J Neurosci 2012, 32:10383-10395.
• [29]Lalancette-Hébert M, Julien C, Cordeau P, Bohacek I, Weng YC, Calon F, Kriz J: Accumulation of dietary docosahexaenoic acid in the brain attenuates acute immune response and development of postischemic neuronal damage. Stroke 2011, 42:2903-2909.
• [30]Hill JK, Gunion-Rinker L, Kulhanek D, Lessov N, Kim S, Clark WM, Dixon MP, Nishi R, Stenzel-Poore MP, Eckenstein FP: Temporal modulation of cytokine expression following focal cerebral ischemia in mice. Brain Res 1999, 820:45-54.
• [31]Ladeby R, Wirenfeldt M, Garcia-Ovejero D, Fenger C, Dissing-Olesen L, Dalmau I, Finsen B: Microglial cell population dynamics in the injured adult central nervous system. Brain Res Brain Res Rev 2005, 48:196-206.
• [32]Nakajima K, Kohsaka S: Microglia: neuroprotective and neurotrophic cells in the central nervous system. Curr Drug Targets Cardiovasc Haematol Disord 2004, 4:65-84.
• [33]O’Donnell SL, Frederick TJ, Krady JK, Vannucci SJ, Wood TL: IGF-I and microglia/macrophage proliferation in the ischemic mouse brain. Glia 2002, 39:85-97.
• [34]Microglia: active sensor and versatile effector cells in the normal and pathologic brain Nat Neurosci 2007, 10:1387-1394.
• [35]Laflamme N, Echchannaoui H, Landmann R, Rivest S: Cooperation between toll-like receptor 2 and 4 in the brain of mice challenged with cell wall components derived from gram-negative and gram-positive bacteria. Eur J Immunol 2003, 33:1127-1138.
• [36]Takeda K, Akira S: Toll receptors and pathogen resistance. Cell Microbiol 2003, 5:143-153.
• [37]Akira S, Uematsu S, Takeuchi O: Pathogen recognition and innate immunity. Cell 2006, 124:783-801.
• [38]Olson JK, Miller SD: Microglia initiate central nervous system innate and adaptive immune responses through multiple TLRs. J Immunol 2004, 173:3916-3924.
• [39]Jack CS, Arbour N, Manusow J, Montgrain V, Blain M, McCrea E, Shapiro A, Antel JP: TLR signaling tailors innate immune responses in human microglia and astrocytes. J Immunol 2005, 175:4320-4330.
• [40]Kielian T: Toll-like receptors in central nervous system glial inflammation and homeostasis. J Neurosci Res 2006, 883:711-730.
• [41]Rolls A, Schechter R, London A, Ziv Y, Ronen A, Levy R, Schwartz M: Toll-like receptors modulate adult hippocampal neurogenesis. Nat Cell Biol 2007, 9:1081-1088.
• [42]Hoffmann O, Braun JS, Becker D, Halle A, Freyer D, Dagand E, Lehnardt S, Weber JR: TLR2 mediates neuroinflammation and neuronal damage. J Immunol 2007, 178:6476-6481.
• [43]Kriz J: Inflammation in ischemic brain injury: timing is important. Crit Rev Neurobiol 2006, 18:145-157.
• [44]Stoll G, Jander S, Schroeter M: Inflammation and glial responses in ischemic brain lesions. Prog Neurobiol 1998, 56:149-171.
• [45]Raivich G, Bohatshek M, Kloss CU, Werner A, Jones LL, Kreutzberg GW: Neuroglial activation repertoire in the injured brain: graded response, molecular mechanisms and cues to physiological function. Brain Res Brain Res Rev 1999, 30:77-105.
• [46]Hailer NP, Grampp A, Nitsch R: Proliferation of microglia and astrocytes in the dentate gyrus following enthorinal cortex lesion: a quantitiative brodmodeoxyuridine labelling study. Eur J Neurosci 1999, 11:3359-3364.
• [47]Bechmann I, Goldmann J, Kovac AD, Kwidzinski E, Simburger E, Naftolin F, Dirnagl U, Nitsch R, Priller J: Circulating monocytic cells infiltrate layers of anterograde axonal degeneration where they transform into microglia. FASEB J 2005, 15:1086-1088.
• [48]Wirenfeldt M, Babcock AA, Ladeby R, Lambertsen KL, Dagnaes-Hansen F, Leslie RG, Owens T, Finsen B: Reactive microgliosis engages distinct responses by microglial subpopulations after minor central nervous system injury. J Neurosci Res 2005, 82:507-514.
• [49]Poon IK, Hulett MD, Parish CR: Molecular mechanisms of late apoptotic/necrotic cell clearance. Cell Death Differ 2010, 17:381-397.