Neurocritical Care

, Volume 12, Issue 2, pp 274–284 | Cite as

CNS Immune Responses Following Experimental Stroke

  • Dannielle Zierath
  • Matthew Thullbery
  • Jessica Hadwin
  • J. Michael Gee
  • Anna Savos
  • Angela Kalil
  • Kyra J. Becker
Translational Research


Background and purpose

Animals subjected to an inflammatory insult with lipopolysaccharide (LPS) at the time of stroke are predisposed to develop a detrimental autoimmune response to myelin basic protein (MBP). In this study, we sought to determine whether other inflammatory stimuli could similarly invoke central nervous system (CNS) autoimmunity and whether these detrimental autoimmune responses occurred to antigens other than MBP.


Male Lewis rats underwent 3 h middle cerebral artery occlusion (MCAO) and received intraperitoneal injections of LPS, staphylococcal enterotoxin B (SEB), lipoteichoic acid (LTA) or saline at the time of reperfusion. Behavioral tests were performed at set time intervals after MCAO and animals were sacrificed at 1 month to analyze the immune response to MBP, neuron specific enolase (NSE) and proteolipid protein (PLP).


Lymphocytes from SEB treated animals were highly reactive to all tested CNS antigens, but treatment with LPS was most likely to lead to a Th1(+) response. A Th1(+) response to MBP, NSE or PLP in spleen was associated with worse outcome, although the response to NSE was most predictive of poor outcome. Animals with a cell mediated autoimmune response to either MBP or NSE in spleen had a concomitant humoral response to these antigens.


These data show that LPS, but not other inflammatory stimuli, increase the likelihood of developing a detrimental autoimmune response to an array of brain antigens.


Stroke Toll-like receptor Autoimmune LPS LTA SEB MBP NSE PLP ThFractalkine 


  1. 1.
    Netea MG, van der Graaf C, Van der Meer JW, Kullberg BJ. Toll-like receptors and the host defense against microbial pathogens: bringing specificity to the innate-immune system. J Leukoc Biol. 2004;75:749–55.CrossRefPubMedGoogle Scholar
  2. 2.
    Janeway CA Jr, Medzhitov R. Innate immune recognition. Annu Rev Immunol. 2002;20:197–216.CrossRefPubMedGoogle Scholar
  3. 3.
    Becker KJ, Kindrick DL, Lester MP, Shea C, Ye ZC. Sensitization to brain antigens after stroke is augmented by lipopolysaccharide. J Cereb Blood Flow Metab. 2005;25:1634–44.CrossRefPubMedGoogle Scholar
  4. 4.
    Kilic U, Kilic E, Matter CM, Bassetti CL, Hermann DM. TLR-4 deficiency protects against focal cerebral ischemia and axotomy-induced neurodegeneration. Neurobiol Dis. 2008;31:33–40.CrossRefPubMedGoogle Scholar
  5. 5.
    Caso JR, Pradillo JM, Hurtado O, Leza JC, Moro MA, Lizasoain I. Toll-like receptor 4 is involved in subacute stress-induced neuroinflammation and in the worsening of experimental stroke. Stroke. 2008;39:1314–20.CrossRefPubMedGoogle Scholar
  6. 6.
    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;353:509–14.CrossRefPubMedGoogle Scholar
  7. 7.
    Aslanyan S, Weir CJ, Diener HC, Kaste M, Lees KR. Pneumonia and urinary tract infection after acute ischaemic stroke: a tertiary analysis of the GAIN International trial. Eur J Neurol. 2004;11:49–53.CrossRefPubMedGoogle Scholar
  8. 8.
    Harms H, Prass K, Meisel C, et al. Preventive antibacterial therapy in acute ischemic stroke: a randomized controlled trial. PLoS ONE. 2008;3:e2158.CrossRefPubMedGoogle Scholar
  9. 9.
    Longa EZ, Weinstein PR, Carlson S, Cummins R. Reversible middle cerebral artery occlusion without craniectomy in rats. Stroke. 1989;20:84–91.PubMedGoogle Scholar
  10. 10.
    Huang W, Koller LD. Superantigen activation and kinetics of cytokines in the Long-Evans rat. Immunology. 1998;95:331–8.CrossRefPubMedGoogle Scholar
  11. 11.
    Chatterjee PK, Zacharowski K, Cuzzocrea S, et al. Lipoteichoic acid from Staphylococcus aureus reduces renal ischemia/reperfusion injury. Kidney Int. 2002;62:1249–63.CrossRefPubMedGoogle Scholar
  12. 12.
    Bederson JB, Pitts LH, Tsuji M, Nishimura MC, Davis RL, Bartkowski H. Rat middle cerebral artery occlusion: evaluation of the model and development of a neurologic examination. Stroke. 1986;17:472–6.PubMedGoogle Scholar
  13. 13.
    Hernandez TD, Schallert T. Seizures and recovery from experimental brain damage. Exp Neurol. 1988;102:318–24.CrossRefPubMedGoogle Scholar
  14. 14.
    Becker K, Kindrick D, McCarron R, Hallenbeck J, Winn R. Adoptive transfer of myelin basic protein-tolerized splenocytes to naive animals reduces infarct size: a role for lymphocytes in ischemic brain injury? Stroke. 2003;34:1809–15.CrossRefPubMedGoogle Scholar
  15. 15.
    Becker KJ. Sensitization and tolerization to brain antigens in stroke. Neuroscience. 2009;158:1090–7.CrossRefPubMedGoogle Scholar
  16. 16.
    Pasare C, Medzhitov R. Toll-like receptors and acquired immunity. Semin Immunol. 2004;16:23–6.CrossRefPubMedGoogle Scholar
  17. 17.
    Toubi E, Shoenfeld Y. Toll-like receptors and their role in the development of autoimmune diseases. Autoimmunity. 2004;37:183–8.CrossRefPubMedGoogle Scholar
  18. 18.
    Huang YH, Haegerstrand A, Frostegard J. Effects of in vitro hyperthermia on proliferative responses and lymphocyte activity. Clin Exp Immunol. 1996;103:61–6.CrossRefPubMedGoogle Scholar
  19. 19.
    Croft M, Dubey C. Accessory molecule and costimulation requirements for CD4 T cell response. Crit Rev Immunol. 1997;17:89–118.PubMedGoogle Scholar
  20. 20.
    Hickey WF, Kimura H. Perivascular microglial cells of the CNS are bone marrow-derived and present antigen in vivo. Science. 1988;239:290–2.CrossRefPubMedGoogle Scholar
  21. 21.
    Kato H, Kogure K, Liu XH, Araki T, Itoyama Y. Progressive expression of immunomolecules on activated microglia and invading leukocytes following focal cerebral ischemia in the rat. Brain Res. 1996;734:203–12.CrossRefPubMedGoogle Scholar
  22. 22.
    Hathcock KS, Laszlo G, Pucillo C, Linsley P, Hodes RJ. Comparative analysis of B7-1 and B7-2 costimulatory ligands: expression and function. J Exp Med. 1994;180:631–40.CrossRefPubMedGoogle Scholar
  23. 23.
    Jiang-Shieh YF, Yeh KY, Wei IH, et al. Responses of microglia in vitro to the gram-positive bacterial component, lipoteichoic acid. J Neurosci Res. 2005;82:515–24.CrossRefPubMedGoogle Scholar
  24. 24.
    Lee SJ, Lee S. Toll-like receptors and inflammation in the CNS. Curr Drug Targets Inflamm Allergy. 2002;1:181–91.CrossRefPubMedGoogle Scholar
  25. 25.
    Zanin-Zhorov A, Tal-Lapidot G, Cahalon L, et al. Cutting edge: T cells respond to lipopolysaccharide innately via TLR4 signaling. J Immunol. 2007;179:41–4.PubMedGoogle Scholar
  26. 26.
    Manicassamy S, Ravindran R, Deng J, et al. Toll-like receptor 2-dependent induction of vitamin A-metabolizing enzymes in dendritic cells promotes T regulatory responses and inhibits autoimmunity. Nat Med. 2009;15:401–9.CrossRefPubMedGoogle Scholar
  27. 27.
    Kerfoot SM, Long EM, Hickey MJ, et al. TLR4 contributes to disease-inducing mechanisms resulting in central nervous system autoimmune disease. J Immunol. 2004;173:7070–7.PubMedGoogle Scholar
  28. 28.
    Kato N, Fujii Y, Agata N, et al. Experimental murine model for autoimmune myocarditis using Klebsiella pneumoniae O3 lipopolysaccharide as a potent immunological adjuvant. Autoimmunity. 1993;14:231–6.CrossRefPubMedGoogle Scholar
  29. 29.
    Zaccone P, Fehervari Z, Blanchard L, Nicoletti F, Edwards CK 3rd, Cooke A. Autoimmune thyroid disease induced by thyroglobulin and lipopolysaccharide is inhibited by soluble TNF receptor type I. Eur J Immunol. 2002;32:1021–8.CrossRefPubMedGoogle Scholar
  30. 30.
    Proft T, Fraser JD. Bacterial superantigens. Clin Exp Immunol. 2003;133:299–306.CrossRefPubMedGoogle Scholar
  31. 31.
    Friedman SM, Tumang JR, Crow MK. Microbial superantigens as etiopathogenic agents in autoimmunity. Rheum Dis Clin North Am. 1993;19:207–22.PubMedGoogle Scholar
  32. 32.
    Ivars F. Superantigen-induced regulatory T cells in vivo. Chem Immunol Allergy. 2007;93:137–60.CrossRefPubMedGoogle Scholar
  33. 33.
    Romagnani S. Regulation of the T cell response. Clin Exp Allergy. 2006;36:1357–66.CrossRefPubMedGoogle Scholar
  34. 34.
    Gee JM, Kalil A, Thullbery M, Becker KJ. Induction of immunologic tolerance to myelin basic protein prevents central nervous system autoimmunity and improves outcome after stroke. Stroke. 2008;39:1575–82.CrossRefPubMedGoogle Scholar
  35. 35.
    Hofstetter HH, Targoni OS, Karulin AY, Forsthuber TG, Tary-Lehmann M, Lehmann PV. Does the frequency and avidity spectrum of the neuroantigen-specific T cells in the blood mirror the autoimmune process in the central nervous system of mice undergoing experimental allergic encephalomyelitis? J Immunol. 2005;174:4598–605.PubMedGoogle Scholar
  36. 36.
    Muhallab S, Lidman O, Weissert R, Olsson T, Svenningsson A. Intra-CNS activation by antigen-specific T lymphocytes in experimental autoimmune encephalomyelitis. J Neuroimmunol. 2001;113:202–11.CrossRefPubMedGoogle Scholar
  37. 37.
    Wang WZ, Olsson T, Kostulas V, Hojeberg B, Ekre HP, Link H. Myelin antigen reactive T cells in cerebrovascular diseases. Clin Exp Immunol. 1992;88:157–62.PubMedCrossRefGoogle Scholar
  38. 38.
    Bornstein NM, Aronovich B, Korczyn AD, Shavit S, Michaelson DM, Chapman J. Antibodies to brain antigens following stroke. Neurology. 2001;56:529–30.PubMedGoogle Scholar
  39. 39.
    Dambinova SA, Khounteev GA, Izykenova GA, Zavolokov IG, Ilyukhina AY, Skoromets AA. Blood test detecting autoantibodies to N-methyl-d-aspartate neuroreceptors for evaluation of patients with transient ischemic attack and stroke. Clin Chem. 2003;49:1752–62.CrossRefPubMedGoogle Scholar
  40. 40.
    Dale RC, Candler PM, Church AJ, Wait R, Pocock JM, Giovannoni G. Neuronal surface glycolytic enzymes are autoantigen targets in post-streptococcal autoimmune CNS disease. J Neuroimmunol. 2006;172:187–97.CrossRefPubMedGoogle Scholar
  41. 41.
    Fillit HM, Kemeny E, Luine V, Weksler ME, Zabriskie JB. Antivascular antibodies in the sera of patients with senile dementia of the Alzheimer’s type. J Gerontol. 1987;42:180–4.PubMedGoogle Scholar
  42. 42.
    Jankovic BD, Horvat J, Djordjijevic D, Ramah A, Fridman V, Spahic O. Brain-associated autoimmune features in heroin addicts: correlation to HIV infection and dementia. Int J Neurosci. 1991;58:113–26.CrossRefPubMedGoogle Scholar
  43. 43.
    Braus BK, Hauck SM, Amann B, et al. Neuron-specific enolase antibodies in patients with sudden acquired retinal degeneration syndrome. Vet Immunol Immunopathol. 2008;124:177–83.CrossRefPubMedGoogle Scholar
  44. 44.
    Ikeda Y, Maruyama I, Nakazawa M, Ohguro H. Clinical significance of serum antibody against neuron-specific enolase in glaucoma patients. Jpn J Ophthalmol. 2002;46:13–7.CrossRefPubMedGoogle Scholar
  45. 45.
    Jankovic BD, Djordjijevic D. Differential appearance of autoantibodies to human brain S100 protein, neuron specific enolase and myelin basic protein in psychiatric patients. Int J Neurosci. 1991;60:119–27.CrossRefPubMedGoogle Scholar
  46. 46.
    Dantzer R. Cytokine, sickness behavior, and depression. Immunol Allergy Clin North Am. 2009;29:247–64.CrossRefPubMedGoogle Scholar
  47. 47.
    Imai T, Hieshima K, Haskell C, et al. Identification and molecular characterization of fractalkine receptor CX3CR1, which mediates both leukocyte migration and adhesion. Cell. 1997;91:521–30.CrossRefPubMedGoogle Scholar
  48. 48.
    Umehara H, Bloom E, Okazaki T, Domae N, Imai T. Fractalkine and vascular injury. Trends Immunol. 2001;22:602–7.CrossRefPubMedGoogle Scholar
  49. 49.
    Chapman GA, Moores K, Harrison D, Campbell CA, Stewart BR, Strijbos PJ. Fractalkine cleavage from neuronal membranes represents an acute event in the inflammatory response to excitotoxic brain damage. J Neurosci. 2000;20:RC87.PubMedGoogle Scholar
  50. 50.
    Tarozzo G, Campanella M, Ghiani M, Bulfone A, Beltramo M. Expression of fractalkine and its receptor, CX3CR1, in response to ischaemia-reperfusion brain injury in the rat. Eur J Neurosci. 2002;15:1663–8.CrossRefPubMedGoogle Scholar
  51. 51.
    Soriano SG, Amaravadi LS, Wang YF, et al. Mice deficient in fractalkine are less susceptible to cerebral ischemia-reperfusion injury. J Neuroimmunol. 2002;125:59–65.CrossRefPubMedGoogle Scholar
  52. 52.
    Denes A, Ferenczi S, Halasz J, Kornyei Z, Kovacs KJ. Role of CX3CR1 (fractalkine receptor) in brain damage and inflammation induced by focal cerebral ischemia in mouse. J Cereb Blood Flow Metab. 2008;28:1707–21.CrossRefPubMedGoogle Scholar
  53. 53.
    Kastenbauer S, Koedel U, Wick M, Kieseier BC, Hartung HP, Pfister HW. CSF and serum levels of soluble fractalkine (CX3CL1) in inflammatory diseases of the nervous system. J Neuroimmunol. 2003;137:210–7.CrossRefPubMedGoogle Scholar
  54. 54.
    Matsunawa M, Isozaki T, Odai T, et al. Increased serum levels of soluble fractalkine (CX3CL1) correlate with disease activity in rheumatoid vasculitis. Arthritis Rheum. 2006;54:3408–16.CrossRefPubMedGoogle Scholar
  55. 55.
    Yajima N, Kasama T, Isozaki T, et al. Elevated levels of soluble fractalkine in active systemic lupus erythematosus: potential involvement in neuropsychiatric manifestations. Arthritis Rheum. 2005;52:1670–5.CrossRefPubMedGoogle Scholar
  56. 56.
    Fraticelli P, Sironi M, Bianchi G, et al. Fractalkine (CX3CL1) as an amplification circuit of polarized Th1 responses. J Clin Invest. 2001;107:1173–81.CrossRefPubMedGoogle Scholar

Copyright information

© Humana Press Inc. 2009

Authors and Affiliations

  • Dannielle Zierath
    • 1
  • Matthew Thullbery
    • 1
  • Jessica Hadwin
    • 1
  • J. Michael Gee
    • 1
  • Anna Savos
    • 1
  • Angela Kalil
    • 1
  • Kyra J. Becker
    • 1
  1. 1.Department of NeurologyUniversity of Washington School of Medicine, Harborview Medical CenterSeattleUSA

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