This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Slifka, M. K.
Right arrow Articles by Whitton, J. L.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Slifka, M. K.
Right arrow Articles by Whitton, J. L.

 Previous Article  |  Next Article 

Journal of Virology, March 2001, p. 2377-2387, Vol. 75, No. 5
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.5.2377-2387.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Using Recombinant Coxsackievirus B3 To Evaluate the Induction and Protective Efficacy of CD8+ T Cells during Picornavirus Infection†

Mark K. Slifka, Robb Pagarigan, Ignacio Mena,Dagger Ralph Feuer, and J. Lindsay Whitton*

Department of Neuropharmacology, The Scripps Research Institute, La Jolla, California

Received 15 September 2000/Accepted 29 November 2000

Coxsackievirus B3 (CVB3) is a common human pathogen that has been associated with serious diseases including myocarditis and pancreatitis. To better understand the effect of cytotoxic T-lymphocyte (CTL) responses in controlling CVB3 infection, we have inserted well-characterized CTL epitopes into the CVB3 genome. Constructs were made by placing the epitope of interest upstream of the open reading frame encoding the CVB3 polyprotein, separated by a poly-glycine linker and an artificial 3Cpro/3CDpro cleavage site. This strategy results in the foreign protein being translated at the amino- terminus of the viral polyprotein, from which it is cleaved prior to viral assembly. In this study, we cloned major histocompatibility complex class I-restricted CTL epitopes from lymphocytic choriomeningitis virus (LCMV) into recombinant CVB3 (rCVB3). In vitro, rCVB3 growth kinetics showed a 1- to 2-h lag period before exponential growth was initiated, and peak titers were ~1 log unit lower than for wild-type virus. rCVB3 replicated to high titers in vivo and caused severe pancreatitis but minimal myocarditis. Despite the high virus titers, rCVB3 infection of naive mice failed to induce a strong CD8+ T-cell response to the encoded epitope; this has implications for the proposed role of "cross-priming" during virus infection and for the utility of recombinant picornaviruses as vaccine vectors. In contrast, rCVB3 infection of LCMV-immune mice resulted in direct ex vivo cytotoxic activity against target cells coated with the epitope peptide, demonstrating that the rCVB3-encoded LCMV-specific epitope was expressed and presented in vivo. The preexisting CD8+ memory T cells could limit rCVB replication; compared to naive mice, infection of LCMV-immune mice with rCVB3 resulted in ~50-fold-lower virus titers in the heart and ~6-fold-lower virus titers in the pancreas. Although the inserted CTL epitope was retained by rCVB3 through several passages in tissue culture, it was lost in an organ-specific manner in vivo; a substantial proportion of viruses from the pancreas retained the insert, compared to only 0 to 1.8% of myocardial viruses. Together, these results show that expression of heterologous viral proteins by recombinant CVB3 provides a useful model for determining the mechanisms underlying the immune response to this viral pathogen.

* Corresponding author. Mailing address: Department of Neuropharmacology, CVN-9, The Scripps Research Institute, 10550 N. Torrey Pines Rd., La Jolla, CA 92037. Phone: (858) 784-7090. Fax: (858) 784-7380. E-mail: lwhitton{at}scripps.edu.

dagger Manuscript 12836-NP from the Scripps Research Institute.

Dagger Present address: Institut Pasteur, 75724 Paris Cedex 15, France.

Journal of Virology, March 2001, p. 2377-2387, Vol. 75, No. 5
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.5.2377-2387.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Feuer, R., Ruller, C. M., An, N., Tabor-Godwin, J. M., Rhoades, R. E., Maciejewski, S., Pagarigan, R. R., Cornell, C. T., Crocker, S. J., Kiosses, W. B., Pham-Mitchell, N., Campbell, I. L., Whitton, J. L. (2009). Viral Persistence and Chronic Immunopathology in the Adult Central Nervous System following Coxsackievirus Infection during the Neonatal Period. J. Virol. 83: 9356-9369 [Abstract] [Full Text]  
  • Marchant, D., Sall, A., Si, X., Abraham, T., Wu, W., Luo, Z., Petersen, T., Hegele, R. G., McManus, B. M. (2009). ERK MAP kinase-activated Arf6 trafficking directs coxsackievirus type B3 into an unproductive compartment during virus host-cell entry. J. Gen. Virol. 90: 854-862 [Abstract] [Full Text]  
  • Weinzierl, A. O., Rudolf, D., Maurer, D., Wernet, D., Rammensee, H.-G., Stevanovic, S., Klingel, K. (2008). Identification of HLA-A*01- and HLA-A*02-restricted CD8+ T-cell epitopes shared among group B enteroviruses. J. Gen. Virol. 89: 2090-2097 [Abstract] [Full Text]  
  • Kemball, C. C., Harkins, S., Whitton, J. L. (2008). Enumeration and Functional Evaluation of Virus-Specific CD4+ and CD8+ T Cells in Lymphoid and Peripheral Sites of Coxsackievirus B3 Infection. J. Virol. 82: 4331-4342 [Abstract] [Full Text]  
  • Crocker, S. J., Frausto, R. F., Whitmire, J. K., Benning, N., Milner, R., Whitton, J. L. (2007). Amelioration of Coxsackievirus B3-Mediated Myocarditis by Inhibition of Tissue Inhibitors of Matrix Metalloproteinase-1. Am. J. Pathol. 171: 1762-1773 [Abstract] [Full Text]  
  • Cornell, C. T., Kiosses, W. B., Harkins, S., Whitton, J. L. (2007). Coxsackievirus B3 Proteins Directionally Complement Each Other To Downregulate Surface Major Histocompatibility Complex Class I. J. Virol. 81: 6785-6797 [Abstract] [Full Text]  
  • Cornell, C. T., Kiosses, W. B., Harkins, S., Whitton, J. L. (2006). Inhibition of protein trafficking by coxsackievirus b3: multiple viral proteins target a single organelle.. J. Virol. 80: 6637-6647 [Abstract] [Full Text]  
  • Fujinami, R. S., von Herrath, M. G., Christen, U., Whitton, J. L. (2006). Molecular Mimicry, Bystander Activation, or Viral Persistence: Infections and Autoimmune Disease. Clin. Microbiol. Rev. 19: 80-94 [Abstract] [Full Text]  
  • Harkins, S., Cornell, C. T., Whitton, J. L. (2005). Analysis of Translational Initiation in Coxsackievirus B3 Suggests an Alternative Explanation for the High Frequency of R+4 in the Eukaryotic Consensus Motif. J. Virol. 79: 987-996 [Abstract] [Full Text]  
  • Chua, M. M., MacNamara, K. C., San Mateo, L., Shen, H., Weiss, S. R. (2004). Effects of an Epitope-Specific CD8+ T-Cell Response on Murine Coronavirus Central Nervous System Disease: Protection from Virus Replication and Antigen Spread and Selection of Epitope Escape Mutants. J. Virol. 78: 1150-1159 [Abstract] [Full Text]  
  • Feuer, R., Mena, I., Pagarigan, R. R., Harkins, S., Hassett, D. E., Whitton, J. L. (2003). Coxsackievirus B3 and the Neonatal CNS: The Roles of Stem Cells, Developing Neurons, and Apoptosis in Infection, Viral Dissemination, and Disease. Am. J. Pathol. 163: 1379-1393 [Abstract] [Full Text]  
  • Kim, T. S., Perlman, S. (2003). Protection Against CTL Escape and Clinical Disease in a Murine Model of Virus Persistence. J. Immunol. 171: 2006-2013 [Abstract] [Full Text]  
  • Feuer, R., Mena, I., Pagarigan, R., Slifka, M. K., Whitton, J. L. (2002). Cell Cycle Status Affects Coxsackievirus Replication, Persistence, and Reactivation In Vitro. J. Virol. 76: 4430-4440 [Abstract] [Full Text]  
  • Lee, S.-G., Kim, D.-Y., Hyun, B.-H., Bae, Y.-S. (2002). Novel Design Architecture for Genetic Stability of Recombinant Poliovirus: the Manipulation of G/C Contents and Their Distribution Patterns Increases the Genetic Stability of Inserts in a Poliovirus-Based RPS-Vax Vector System. J. Virol. 76: 1649-1662 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»