Tick-borne encephalitis (TBE) virus causes severe encephalitis with serious sequelae in humans. The disease is characterized by fever and debilitating encephalitis that can progress to chronic illness or fatal infection. In this study, changes in permeability of the blood-brain barrier (BBB) in two susceptible animal models (BALB/c, and C57Bl/6 mice) infected with TBE virus were investigated at various days after infection by measuring fluorescence in brain homogenates after intraperitoneal injection of sodium fluorescein, a compound that is normally excluded from the central nervous system. We demonstrate here that TBE virus infection, in addition to causing fatal encephalitis in mice, induces considerable breakdown of the BBB. The permeability of the BBB increased at later stages of TBE infection when high virus load was present in the brain (i.e., BBB breakdown was not necessary for TBE virus entry into the brain), and at the onset of the first severe clinical symptoms of the disease, which included neurological signs associated with sharp declines in body weight and temperature. The increased BBB permeability was in association with dramatic upregulation of proinflammatory cytokine/chemokine mRNA expression in the brain. Breakdown of the BBB was also observed in mice deficient in CD8(+) T-cells, indicating that these cells are not necessary for the increase in BBB permeability that occurs during TBE. These novel findings are highly relevant to the development of future therapies designed to control this important human infectious disease.

Institute of Parasitology Biology Centre of the Czech Academy of Sciences České Budějovice Czech Republic

Zobrazit více v PubMed

Andersen IH, Marker O, Thomsen AR. Breakdown of blood-brain barrier function in the murine lymphocytic choriomeningitis virus infection mediated by virus-specific CD8+ T cells. J Neuroimmunol. 1991;31:155–163. PubMed

Kanmogne GD, Primeaux C, Grammas P. HIV-1 gp120 proteins alter tight junction protein expression and brain endothelial cell permeability: implications for the pathogenesis of HIV-associated dementia. J Neuropathol Exp Neurol. 2005;64:498–505. PubMed

Morgan L, Shah B, Rivers LE, Barden L, Groom AJ, et al. Inflammation and dephosphorylation of the tight junction protein occludin in an experimental model of multiple sclerosis. Neuroscience. 2007;147:664–673. PubMed

Olsen AL, Morrey JD, Smee DF, Sidwell RW. Correlation between breakdown of the blood-brain barrier and disease outcome of viral encephalitis in mice. Antiviral Res. 2007;75:104–112. PubMed PMC

Gralinski LE, Ashley SL, Dixon SD, Spindler KR. Mouse adenovirus type 1-induced breakdown of the blood-brain barrier. J Virol. 2009;83:9398–9410. PubMed PMC

Silver NC, Tofts PS, Symms MR, Barker GJ, Thompson AJ, et al. Quantitative contrast-enhanced magnetic resonance imaging to evaluate blood-brain barrier integrity in multiple sclerosis: a preliminary study. Mult Scler. 2001;7:75–82. PubMed

Hooper DC, Scott GS, Zborek A, Mikheeva T, Kean RB, et al. Uric acid, a peroxynitrite scavenger, inhibits CNS inflammation, blood-CNS barrier permeability changes, and tissue damage in a mouse model of multiple sclerosis. FASEB J. 2000;14:691–698. PubMed

Hooper DC, Kean RB, Scott GS, Spitsin SV, Mikheeva T, et al. The central nervous system inflammatory response to neurotropic virus infection is peroxynitrite dependent. J Immunol. 2001;167:3470–3477. PubMed

Phares TW, Kean RB, Mikheeva T, Hooper DC. Regional differences in blood-brain barrier permeability changes and inflammation in the apathogenic clearance of virus from the central nervous system. J Immunol. 2006;176:7666–7675. PubMed

Wang T, Town T, Alexopoulou L, Anderson JF, Fikrig E, et al. Toll-like receptor 3 mediates West Nile virus entry into the brain causing lethal encephalitis. Nat Med. 2004;10:1366–1373. PubMed

Liu TH, Liang LC, Wang CC, Liu HC, Chen WJ. The blood-brain barrier in the cerebrum is the initial site for the Japanese encephalitis virus entering the central nervous system. J Neurovirol. 2008;14:514–521. PubMed

Banks WA, Ercal N, Price TO. The blood-brain barrier in neuroAIDS. Curr HIV Res. 2006;4:259–266. PubMed

Süss J. Tick-borne encephalitis in Europe and beyond–the epidemiological situation as of 2007. Euro Surveill. 2008;13 PubMed

Dörrbecker B, Dobler G, Spiegel M, Hufert FT. Tick-borne encephalitis virus and the immune response of the mammalian host. Travel Med Infect Dis. 2010;8:213–222. PubMed

Růžek D, Dobler G, Mantke OD. Tick-borne encephalitis: pathogenesis and clinical implications. Travel Med Infect Dis. 2010;8:223–232. PubMed

Atrasheuskaya AV, Fredeking TM, Ignatyev GM. Changes in immune parameters and their correction in human cases of tick-borne encephalitis. Clin Exp Immunol. 2003;131:148–154. PubMed PMC

Campbell IL, Abraham CR, Masliah E, Kemper P, Inglis JD, et al. Neurologic disease induced in transgenic mice by cerebral overexpression of interleukin 6. Proc Natl Acad Sci U S A. 1993;90:10061–10065. PubMed PMC

Beliaeva IA, Antonova OM, Anin AN, Chekhonin VP. [Resistance of the hemato-encephalic barrier in tick-borne neural infection (Lyme disease and tick-borne encephalitis)]. Zh Nevrol Psikhiatr Im S S Korsakova. 1995;95:25–29. PubMed

Chekhonin VP, Zhirkov YA, Belyaeva IA, Ryabukhin IA, Gurina OI, et al. Serum time course of two brain-specific proteins, alpha(1) brain globulin and neuron-specific enolase, in tick-born encephalitis and Lyme disease. Clin Chim Acta. 2002;320:117–125. PubMed

Thach DC, Kimura T, Griffin DE. Differences between C57BL/6 and BALB/cBy mice in mortality and virus replication after intranasal infection with neuroadapted Sindbis virus. J Virol. 2000;74:6156–6161. PubMed PMC

Tigabu B, Juelich T, Bertrand J, Holbrook MR. Clinical evaluation of highly pathogenic tick-borne flavivirus infection in the mouse model. J Med Virol. 2009;81:1261–1269. PubMed

Morrey JD, Olsen AL, Siddharthan V, Motter NE, Wang H, et al. Increased blood-brain barrier permeability is not a primary determinant for lethality of West Nile virus infection in rodents. J Gen Virol. 2008;89:467–473. PubMed

Pulzova L, Bhide MR, Andrej K. Pathogen translocation across the blood-brain barrier. FEMS Immunol Med Microbiol. 2009;57:203–213. PubMed

Chaturvedi UC, Dhawan R, Khanna M, Mathur A. Breakdown of the blood-brain barrier during dengue virus infection of mice. J Gen Virol. 1991;72(Pt 4):859–866. PubMed

Diamond MS, Klein RS. West Nile virus: crossing the blood-brain barrier. Nat Med. 2004;10:1294–1295. PubMed

Edelmann KH, Richardson-Burns S, Alexopoulou L, Tyler KL, Flavell RA, et al. Does Toll-like receptor 3 play a biological role in virus infections? Virology. 2004;322:231–238. PubMed

Mathur A, Khanna N, Chaturvedi UC. Breakdown of blood-brain barrier by virus-induced cytokine during Japanese encephalitis virus infection. Int J Exp Pathol. 1992;73:603–611. PubMed PMC

Hayasaka D, Nagata N, Fujii Y, Hasegawa H, Sata T, et al. Mortality following peripheral infection with tick-borne encephalitis virus results from a combination of central nervous system pathology, systemic inflammatory and stress responses. Virology. 2009;390:139–150. PubMed

Tigabu B, Juelich T, Holbrook MR. Comparative analysis of immune responses to Russian spring-summer encephalitis and Omsk hemorrhagic fever viruses in mouse models. Virology. 2010;408:57–63. PubMed PMC

Jeohn GH, Kong LY, Wilson B, Hudson P, Hong JS. Synergistic neurotoxic effects of combined treatments with cytokines in murine primary mixed neuron/glia cultures. J Neuroimmunol. 1998;85:1–10. PubMed

Abbott NJ. Inflammatory mediators and modulation of blood-brain barrier permeability. Cell Mol Neurobiol. 2000;20:131–147. PubMed PMC

de Vries HE, Blom-Roosemalen MC, van Oosten M, de Boer AG, van Berkel TJ, et al. The influence of cytokines on the integrity of the blood-brain barrier in vitro. J Neuroimmunol. 1996;64:37–43. PubMed

Yamada M, Kim S, Egashira K, Takeya M, Ikeda T, et al. Molecular mechanism and role of endothelial monocyte chemoattractant protein-1 induction by vascular endothelial growth factor. Arterioscler Thromb Vasc Biol. 2003;23:1996–2001. PubMed

Stamatovic SM, Shakui P, Keep RF, Moore BB, Kunkel SL, et al. Monocyte chemoattractant protein-1 regulation of blood-brain barrier permeability. J Cereb Blood Flow Metab. 2005;25:593–606. PubMed

Michalowska-Wender G, Losy J, Kondrusik M, Zajkowska J, Pancewicz S, et al. [Evaluation of soluble platelet cell adhesion molecule sPECAM-1 and chemokine MCP-1 (CCL2) concentration in CSF of patients with tick-borne encephalitis]. Pol Merkur Lekarski. 2006;20:46–48. PubMed

Grygorczuk S, Zajkowska J, Swierzbinska R, Pancewicz S, Kondrusik M, et al. [Concentration of the beta-chemokine CCL5 (RANTES) in cerebrospinal fluid in patients with tick-borne encephalitis]. Neurol Neurochir Pol. 2006;40:106–111. PubMed

Bolton SJ, Anthony DC, Perry VH. Loss of the tight junction proteins occludin and zonula occludens-1 from cerebral vascular endothelium during neutrophil-induced blood-brain barrier breakdown in vivo. Neuroscience. 1998;86:1245–1257. PubMed

Lou J, Chofflon M, Juillard C, Donati Y, Mili N, et al. Brain microvascular endothelial cells and leukocytes derived from patients with multiple sclerosis exhibit increased adhesion capacity. Neuroreport. 1997;8:629–633. PubMed

Irani DN, Griffin DE. Regulation of T cell responses during central nervous system viral infection. Adv Virus Res. 2001;56:175–197. PubMed PMC

Rouse BT, Sehrawat S. Immunity and immunopathology to viruses: what decides the outcome? Nat Rev Immunol. 2010;10:514–526. PubMed PMC

Růžek D, Salát J, Palus M, Gritsun TS, Gould EA, et al. CD8+ T-cells mediate immunopathology in tick-borne encephalitis. Virology. 2009;384:1–6. PubMed

Gelpi E, Preusser M, Garzuly F, Holzmann H, Heinz FX, et al. Visualization of Central European tick-borne encephalitis infection in fatal human cases. J Neuropathol Exp Neurol. 2005;64:506–512. PubMed

Gelpi E, Preusser M, Laggner U, Garzuly F, Holzmann H, et al. Inflammatory response in human tick-borne encephalitis: analysis of postmortem brain tissue. J Neurovirol. 2006;12:322–327. PubMed

Suidan GL, McDole JR, Chen Y, Pirko I, Johnson AJ. Induction of blood brain barrier tight junction protein alterations by CD8 T cells. PLoS One. 2008;3:e3037. PubMed PMC

Monath TP, Cropp CB, Harrison AK. Mode of entry of a neurotropic arbovirus into the central nervous system. Reinvestigation of an old controversy. Lab Invest. 1983;48:399–410. PubMed

McMinn PC, Dalgarno L, Weir RC. A comparison of the spread of Murray Valley encephalitis viruses of high or low neuroinvasiveness in the tissues of Swiss mice after peripheral inoculation. Virology. 1996;220:414–423. PubMed

Liou ML, Hsu CY. Japanese encephalitis virus is transported across the cerebral blood vessels by endocytosis in mouse brain. Cell Tissue Res. 1998;293:389–394. PubMed

Verma S, Lo Y, Chapagain M, Lum S, Kumar M, et al. West Nile virus infection modulates human brain microvascular endothelial cells tight junction proteins and cell adhesion molecules: Transmigration across the in vitro blood-brain barrier. Virology. 2009;385:425–433. PubMed PMC

Ruzek D, Bell-Sakyi L, Kopecky J, Grubhoffer L. Growth of tick-borne encephalitis virus (European subtype) in cell lines from vector and non-vector ticks. Virus Res. 2008;137:142–146. PubMed

Rey FA, Heinz FX, Mandl C, Kunz C, Harrison SC. The envelope glycoprotein from tick-borne encephalitis virus at 2 A resolution. Nature. 1995;375:291–298. PubMed

De Madrid AT, Porterfield JS. A simple micro-culture method for the study of group B arboviruses. Bull World Health Organ. 1969;40:113–121. PubMed PMC

Babu S, Nutman TB. Proinflammatory cytokines dominate the early immune response to filarial parasites. J Immunol. 2003;171:6723–6732. PubMed

History of Arbovirus Research in the Czech Republic

Changes in cytokine and chemokine profiles in mouse serum and brain, and in human neural cells, upon tick-borne encephalitis virus infection

Electron Tomography Analysis of Tick-Borne Encephalitis Virus Infection in Human Neurons

Mice with different susceptibility to tick-borne encephalitis virus infection show selective neutralizing antibody response and inflammatory reaction in the central nervous system