Abstract
We herein report an investigation of nitric oxide (NO) levels, a candidate molecule for neuronal toxicity and dysfunction, in the brain of rabbits during experimental neurological infection by bovine herpesvirus 5 (BoHV-5). Spectrophotometry for NO products (NO2 and NO3) revealed that NO levels were significantly increased (F(4, 40) = 3.33; P <.02) in several regions of the brain of rabbits with neurological disease, correlating with moderate to high BoHV-5 titers. Immunohistochemistry of brain regions revealed a group of cells with neuronal and astrocyte morphology expressing the enzyme inducible NO synthase (iNOS) close to virus antigenpositive neurons. In addition, the investigation of nitric oxide levels between 2 and 6 days post infection (d.p.i.) revealed an initial increase in NO levels in the olfactory bulb and cortex (OB/OC) and anterior cortex (AC) at day 3 p.i., correlating with the initial detection of virus. As the infection proceeded, increased NO levels— and infectivity—were progressively being detected in the OB/CO and AC at day 4 p.i. (F(12, 128) = 2.82; P <.003); at day 5 p.i. in several brain regions (P <.003 in the OB/OC); and at day 6 p.i. in all regions (P <.003) but the thalamus. These results show that BoHV-5 replication in the brain of rabbits induces an overproduction of NO. The increase in NO levels in early infection correlated spatially and temporally with virus dissemination within the brain and preceded the development of neurological signs. Thus, the overproduction of NO in the brain of BoHV-5-infected rabbits may be a component of the pathogenesis of BoHV-5-induced neurological disease.
Similar content being viewed by others
References
Akaike T, Maeda H (2000). Nitric oxide and virus infection. Immunology 101: 300–308.
Bagetta G, Paoletti AM, Leta A, Del Duca C, Nisticò R, Rotiroti D, Corasaniti MT (2002). Abnormal expression of neuronal nitric oxide synthase triggers limbic seizures and hippocampal damage in rat. Biochem Biophys Res Commun 2: 255–260.
Bogdan, C (1998). The Multiplex Function of Nitric Oxide in (Auto) immunity. J Exp Med 187: 9, 1361–1365.
Bradford MM (1976). A rapid and sensitive method for the quantitation of micrograms quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 7: 248–254.
Carrilo BJ, Pospischil A, Dahme E (1983). Pathology of a bovine viral necrozing encephalitis in Argentina. Zbl Vet Med B 30: 161–168.
Cerqueira NF, Yoshida WB (2002). Óxido nítrico: revisão. Acta Cir Bras 17: 417–423.
Chesler DA, Reiss CS (2002). The role of IFNγ in immune responses to viral infections of the central nervous system. Cytokine Growth Factor Rev 12: 441–454.
Chowdhury SI, Lee BJ, Mosier D, Sur JH, Osório FA, Kennedy G, Weiss ML (1997). Neurophatology of bovine herpesvirus 5 (BHV-5) meningo-encephalitis in a rabbit seizure model. J Comp Pathol 117: 295–310.
Delhon G, Moraes MP, Lu Z, Afonso CL, Flores EF, Weiblen R, Kutish GF, Rock DL (2003). Genome of bovine herpesvirus 5. J Virol 77: 10339–10347.
Diel DG, Fonseca ET, Souza SF, Mazzanti A, Bauermann FV, Weiblen R, Flores EF (2005). Bovine herpesvirus 5 may use the olfactory and trigeminal pathways to invade the central nervous system of rabbits, depending upon the route of inoculation. Braz J Vet Res 25: 164–170.
Flores EF, Donis RO (1995). Isolation and characterization of a bovine cell line resistent to infection with the pestivirus bovine viral diarrhea virus (BVDV). Virology 208: 565–575.
Fuji S, Akaike T, Maeda H (1999). Role of nitric oxide in pathogenesis of herpes simplex virus encephalitis in rats. Virology 256: 203–212.
Goody, RJ, Schittone, SA, Tyler, KL (2005). Reovirus infection of the CNS enhances iNOS expression in areas of virus induced injury. Exp. Neurol 195: 379–390.
Harris N, Buller RML, Karupiah G (1995). Gamma interferon-induced, nitric oxide-mediated inhibition of vaccinia virus replication. J Virol 69: 910–915.
Hooper DC, Kean RB, Scott GS, Spitsin SV, Mikheeva T, Morimoto K, Bette M, Röhrenbeck AM, Dietzschold B, Weihe E (2001). The central nervous system inflammatory response to neurotropic virus infection is peroxynitrite dependent. J Immunol 167: 3470–3477.
Kahrs RF (2001). Infectious bovine rhinotracheitis and infectious pustular vulvovaginitis. In: Viral diseases of cattle, 2nd ed. Kahrs RF (ed). Ames, IA: Iowa State University, pp 159–170.
Karupiah G, Harris, N (1995). Inhibition of viral replication by nitric oxide and its reversal by ferrous sulfate and tricarboxylic acid cycle metabolites. J Exp Med 181: 2171–2179.
Kiechele FL, Malinski T (1993). Nitric oxide: biochemistry, pathophysiology and detection. Am J Clin Pathol 100: 567–575.
Kodukula P, Liu T, Van Rooijen N, Jager MJ, Hendricks RL (1999). Macrophage control of herpes simplex virus type 1 replication in the peripheral nervous system. J Immunol 162: 2895–2905.
Koprowsky H, Zheng YM, Heber-Katz E, Fraser N, Rorke L, Fu ZF, Hanlon C, Dietzschold B. (1993). In vivo expression of inducible nitric oxide synthase in experimentally induced neurologic diseases. Proc Natl Acad Sci U S A 90: 3024–3027.
Marcaccini A, López-Penã M, Bermudez R, Quiroga MI, Guerrero FH, Nieto JM, Aleman N. (2007). Pseudorabies virus induces a rapid up-regulation of nitric oxide synthases in the nervous system of swine. Vet Microbiol 125: 232–243.
Marques CP, Cheeran MC, Palmquist JM, Hu S, Lokensgard JR. (2008). Microglia are the major cellular source of inducible nitric oxide synthase during experimental herpes encephalitis. J NeuroVirol 14: 229–238.
Marques CP, Hu S, Sheng W, Lokensgard JR. (2006). Microglial cells initiate vigorous yet non-protective immune responses during HSV-1 brain infection. Virus Res 121: 1, 1–10.
Meyer G, Lemaire M, Lyaku J (1996). Establishment of a rabbit model for bovine herpesvirus type 5 neurological acute infection. Vet Microbiol 5: 27–40.
Minc-Golomb D, Tsafaty I, Schwartz JP, (1994). Expression of inducible nitric oxide synthase by neurons following exposure to endotoxin and cytokine. Braz J Pharmacol 112: 720–722.
Minc-Golomb D, Yalid G, Tsarfaty H, Resau JH, Schwartz JP (1996). In vivo expression of inducible nitric oxide synthase in cerebellar neurons. J Neurochem 66: 1504–1509.
Miranda KM, Espey MG, Wink DA (2001). A rapid, simple spectrophotometric method for detection of nitrate and nitrite. Nitric Oxide 5: 62–71.
Moro MA, De Alba J, Leza JC, Lorenzo P, Fernandez AP, Bentura ML, Bosca L, Rodrigo J, Lizasoain, I (1998). Neuronal expression of inducible nitric oxide synthase after oxygen and glucose deprivation in rat forebrain slices. Eur J Neurosci 10: 445–456.
Oldoni I, Weiblen R, Inkelmann MA, Flores EF (2004). Production and characterization of monoclonal antibodies to a Brazilian bovine herpesvirus type 5 (BHV-5). Braz J Med Biol Res 37: 213–221.
Persichini T, Cantoni O, Suzuki H, Colasanti M (2006). Cross-talk between constitutive and inducible NO synthase: an update. Antioxid Redox Signal 8: 949–954.
Reed LJ, Muench HA (1938). A simple method of estimating fifty percent endpoints. Am J Hyg 27: 493–497.
Rissi DR, Oliveira FN, Rech RR, Pierezan F, Lemos RAA, Barros CSL (2006). Epidemiology, clinical signs and distribution of the encephalic lesions in cattle affected by meningoencephalitis caused by bovine herpesvirus-5. Braz J Vet Res 26: 123–132.
Royes LFF, Fighera MR, Furian AF, Oliveira MS, Mysklw JC, Fiorenza NG, Mello CF (2005). Involvement of NO in the convulsive behavior and oxidative damage induced by the intrastriatal injection of methylmalonate. Neurosci Lett 376: 115–120.
Saha RN, Pahan K (2006). Regulation of inducible nitric oxide synthase gene in glial cells. Antioxid Redox Signal 8: 929–947.
Serrano F, Enquist LW, Card JP (2002). Pseudorabies virusinduced expression of nitric oxide synthase isoforms. Physiol Behav 77: 557–563.
Silva AM, Flores EF, Weiblen R, Canto MC, Irigoyen LF, Roehe PM, Souza RS (1999). Pathogenesis of meningoencephalitis in rabbits by bovine herpesvirus type-5 (BHV-5). Rev Bras Microbiol 30: 22–31.
Stewart VC, Heales SJ (2003). Nitric oxide-induced mitochondrial dysfunction: implications for neurodegeneration, Free Radic Biol Med 34: 287–303.
Studdert MJ (1989). Bovine encephalitis herpesvirus. Vet Rec 125: 584.
Ubol S, Sukwattanapan C, Maneerat Y (2001). Inducible nitric oxide synthase inhibition delays death of rabies virus-infected mice. J Med Microbiol 50: 238–242.
Vezzani A (2005). Inflammation and epilepsy. Epilepsy Curr 1: 1–6.
Vogel FSF, Caron L, Flores EF, Weiblen R, Winkelmann ER, Mayer SV, Bastos RG (2003). Distribution of bovine herpesvirus type 5 DNA in the central nervous systems of latently, experimentally infected calves. J Clin Microbiol 41: 4512–4520.
Weiblen R, Barros CS, Canabarro TF, Flores IE (1989). Bovine meningoencephalitis from IBR virus. Vet Rec 25: 124, 666–667.
Wong GKT, Marsden PA (1996). Nitric oxide synthases: regulation in disease. Nephrol Dial Transplant 11: 215–220.
Zaki MH, Akuta T, Akaike T. (2005). Nitric oxide-induced nitrative stress involved in microbial pathogenesis. J Pharmacol Sci 98: 117–129.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Dezengrini, R., Weiss, M., Torres, F.D. et al. Bovine herpesvirus 5 induces an overproduction of nitric oxide in the brain of rabbits that correlates with virus dissemination and precedes the development of neurological signs. Journal of NeuroVirology 15, 153–163 (2009). https://doi.org/10.1080/13550280802578067
Received:
Revised:
Accepted:
Issue Date:
DOI: https://doi.org/10.1080/13550280802578067