Bovine herpesvirus type 1 (BHV-1) and equine herpesvirus type 1 (EHV-1) pathogenesis and genetically engineered vaccines against BHV-1 and EHV-1. BHV-1 as a vaccine vector for immunization against other viral diseases in cattle.
1. Role of envelope proteins/glycoproteins in Bovine herpesvirus type 1 (BHV- 1) neuronal transport/neuropathogenesis.
During primary infection, BHV-1 replicates in the nasal epithelium and enters the
sensory nerve endings (axon terminals) of the trigeminal nerve in the nasopharynx.
The nucleocapsids along with tegument structures are then transported retrogradely
(up the axon, towards the cell body) to the trigeminal ganglion (TG) where they establish
life-long latency. Corticosteroid-induced or stress-mediated reactivation of latent
virus usually leads to its anterograde transport (down the axon, away from the cell
body) to the nerve endings in the nasopharynx (maxillary branch of the trigeminal
nerve) and cornea (ophthalmic branch of the trigeminal nerve). This leads to nasal
and ocular virus shedding and subsequent virus transmission to susceptible animals.
Therefore, retrograde and anterograde transport mechanisms are crucial for the maintenance
and transmission of BHV-1 in the susceptible animal population. We have determined
that the gE-, entire gE cytoplasmic tail-, Us9- and Us9 acidic-domain-deleted viruses
have defective anterograde transport (Veterinary Microbiol. 152:270-279; J. Neurovirol.
16:457-465). One of our future research goals is to understand the BHV-1 retrograde
and anterograde neuronal transport mechanisms. To this end our specific objectives
are: i) To identify the BHV-1 tegument protein that interacts with neuronal dynein
protein for the retrograde transport, ii) To further narrow down the region within
the gE cytoplasmic tail that is important for anteorograde axonal transport and iii)
To identify a neuronal protein or proteins that interact directly or indirectly with
the Us9 acidic domain and/or gE cytoplasmic tail sequences and promote kinesin motor-mediated
anterograde axonal transport.
1. Regulation of pathogenicity and immunogenicity by BHV-1 envelope protein UL49.5/ glycoprotein(g)M
BHV-1, like other alpha herpesviruses can evade the immune recognition at early times post-infection. Recently, it was discovered that envelope protein UL49.5 which forms complex with glycoprotein M, interferes with the cellular immune response by interfering with a transporter associated with antigen presentation (TAP) function. The goals of this project have been i) To identify the UL49.5 domain(s) necessary for disrupting TAP function and gM binding, ii) To construct BHV-1 mutant lacking the TAP inhibition domain and determine its immunogenicity and protective efficacy in calves against BHV-1 infection. Our results showed that UL49.5 cytoplasmic tail residues (80-96) and luminal domain residues 30-32 together determine the maximum TAP inhibition and MHC-I down regulation property of the virus (PLoS one 6(10)e25742). Further, we have determined that a BHV-1 mutant virus lacking these sequences induced more rapid onset of virus neutralizing antibody and cellular immune responses in calves than the wild-type strain Cooper (Vet. Immun. and Immunopath. 147: 223-229).
2. Novel bovine herpesvirus type 1 (BHV-1) vectored vaccine against viral infections associated with bovine respiratory disease complex (BRDC).
Bovine respiratory disease complex (BRDC) is a multifactorial disease in cattle that involves initial viral respiratory infection followed by secondary bacterial infection and severe bronchopneumonia. BRDC costs the US cattle industry more than $ 1 billion/year. Bovine herpesvirus type 1 (BHV-1), bovine respiratory syncytial virus (BRSV), bovine viral diarrheal virus (BVDV) and respiratory bovine corona virus (RBCoV) are important viral agents involved in BRDC. Because of their immunosuppressive (BHV-1, BVDV, BRSV) effects on the host and/or higher mutation rates (BVDV, BRSV, RBCoV), current modified live/attenuated and inactivated vaccines are not adequately efficacious against these virus infections. As noted above, we have determined that BHV-1 gE cytoplasmic tail (CT) and Us9 deletion mutant viruses have defective anterograde neuronal transport from trigeminal ganglia neuronal cell bodies to nose after latency reactivation. To develop an improved vaccine against BHV-1, we have incorporated the UL49.5 mutation discussed above, gE-CT and Us9 mutations into a novel BHV-1 vaccine vector (project funded by Elanco/Eli-Lilly Animal Health). In the near future, the vaccine efficacy study of the above BHV-1 vaccine vector is planned. Further, we will construct BHV-1 vectored subunit vaccines expressing BVDV, BRSV, or RBCoV protective antigens and determine their vaccine efficacy and protection against the corresponding wild-type virus as well as against BHV-1 in calves.
1. Chowdhury, S.I., Onderci, M., Bhattacharjee, P.S., Al-Mubarak, A., Weiss, M.L.,. and Zhou, Y.J. (2002). Bovine herpesvirus type 5 (BHV-5) Us9 is essential for BHV-5 neuropathogenesis. J. Virol.76, 3839-3851.
2. Al-Mubarak, A, Zhou, Y., and Chowdhury, S.I . (2004). A glycine rich region in the ecto domain of BHV-5 gE is important for BHV-5 neuropathogenesis. J. Virol. 78, 4806-4816.
3. Al-Mubarak, A. and Chowdhury, S.I.. (2004). In the absence of glycoprotein I (gI), gE determines bovine herpesvirus type 5 neuroinvasiveness and neurovirulence. J. Neurovirology. 10: 233-243.
4. Chowdhury, S.I., S. Mahmood, J. Simon, A. Al-Mubarak and Y. Zhou (2006). The Us9 gene encoded by bovine herpesvirus type 1 (BHV-1) effectively complements a Us9-null bovine herpesvirus 5 (BHV-5) for anterograde transport, neurovirulence and neuroinvasiveness in a rabbit model. J. Virol. 80: 4396-4405.
5. Al-Mubarak-A., J. Simon, M.D. Burton and S.I. Chowdhury (2007). Glycoprotein E(gE) specified by Bovine herpesvirus type5 (BHV-5) enables trans-neuronal virus spread and neurovirulence without being a structural component of enveloped virions. Virology. 365:398-409.
6. Butchi, N.B, Jones, C., Perez.S, Doster.A, and Chowdhury.S.I (2007). Role of Envelope protein Us9 in the anterograde transport of BHV-1 following reactivation in the Trigeminal Ganglia . J. Neurovirology.13:384-388.
7. Jones, C. and S. Chowdhury (2007). A review of the biology of bovine herpesvirus type 1 (BHV-1), its role as a cofactor in the bovine respiratory disease complex, and development of improved vaccines. Animal Health res. Rev.8:187-205.
8. Liu, Z-F., Brum, M, A. Doster, C. Jones and S.I. Chowdhury (2008). A bovine herpesvirus type 1 mutant virus specifying a carboxyl-terminal truncation of glycoprotein E is defective in anterograde neuronal transport in rabbits and calves. J. Virol. 82: 7432-7442.
9. Koppers-Lalic D, Verweij MC, Lipińska AD, Wang Y, Quinten E, Reits EA, Koch J, Loch S, Rezende MM, Daus F, Bieńkowska-Szewczyk K, Osterrieder N, Mettenleiter TC, Heemskerk MH, Tampé R, Neefjes JJ, Chowdhury SI, ing ME, Rijsewijk FA, Wiertz EJ (2008). Varicellovirus UL 49.5 proteins differentially affect the function of the transporter associated with antigen processing, TAP. Plos Pathog. 30;4(5):e1000080
10. Brum M, Coats C., Sangena B.R, Doster.A., Jones, C. and Chowdhury.S.I. (2009). Role of Envelope proteins gE in the anterograde transport of BHV-1 following reactivation in the Trigeminal Ganglia. J. Neuro virology. 15 (2): 196-201.
11. Brum M.C.S., Weiblen R., Flores E.F. and Chowdhury S.I. (2010). Construction and growth properties of bovine herpesvirus type 5 recombinants defective in the glycoprotein E or thymidine kinase gene or both. Brazilian J. Med. Biol. Res. 43: 217-224.
12. Silva S.C., Brum M.C.S., Weiblen R., Flores E.F. and Chowdhury S.I. (2010), A bovine herpesvirus 5 recombinant defective in the thymidine kinase (TK) gene and a double mutant lacking TK and the glycoprotein E gene are fully attenuated for rabbits. Brazilian J. Med. Biol. Res. 43: 150-159.
13. Brum M.C.S., Caron L., Chowdhury S.I. ,Weiblen R., and Flores E.F. (2010). Immunogenicity of an inactivated bovine herpesvirus type 5 strain defective in the thymidine kinase and glycoprotein E. Pesq. Vet. Bras. 30: 57-62.
14. Clinton Jones and Shafiqul Chowdhury (2010). Bovine herpesvirus type 1 (BHV-1) is an important cofactor in the bovine respiratory disease complex. Vet. Clinics of North America: Food Animal Practice, 26:303-321.
15. Chowdhury, S.I., Coats, C.J., Neis, RA., Navarro, S.M., Paulsen,D.B. and Feng, J.M.(2010). A Bovine Herpesvirus Type 1 (BHV-1) mutant virus with truncated glycoprotein E cytoplasmic tail has defective anterograde neuronal transport in rabbit dorsal root ganglionic primary neuronal cultures in a microfluidic chamber system. J. Neurovirol. 16 (6): 457 465.
16. Chowdhury.S.I. Brum M, Coats C., Doster.A. and Jones, C. (2011). A Bovine Herpesvirus Type 1 Mutant Virus lacking Us9 acidic domain sequences is defective in Anterograde axonal transport in calves and in microfluidic chamber cultures of dissociated and compartmentalized rabbit dorsal root neurons.Vet Microbiol.152:270-279.
17. Wei H., Wang, Y.,and Chowdhury, S.I.(2011). Bovine herpesvirus type 1 (BHV-1)
UL49.5 luminal domain residues 30-32 are critical for MHC-I down-regulation in virus-
infected cells. PLoS one 6(10) e25742.
18. Wei H., He J., Paulsen D.B., Chowdhury, S.I. (2012). Bovine herpesvirus type 1
(BHV-1) UL49.5 luminal domain residues 30-32 and cytoplasmic tail residues 80-96 induce
better immune responses in claves than the wild-type strain Cooper. Vet. Immunol. and Immunopathol. 147: 223-229.