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 Chapman, N. M.
Right arrow Articles by Kolbeck, P.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Chapman, N. M.
Right arrow Articles by Kolbeck, P.

 Previous Article  |  Next Article 

Journal of Virology, September 2000, p. 7952-7962, Vol. 74, No. 17
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Coxsackievirus Expression of the Murine Secretory Protein Interleukin-4 Induces Increased Synthesis of Immunoglobulin G1 in Mice

Nora M. Chapman,1,* Kyung-Soo Kim,1 Steven Tracy,1 John Jackson,2 Katja Höfling,1 J. Smith Leser,1 James Malone,2,dagger and Peter Kolbeck2,Dagger

Enterovirus Research Laboratory, Department of Pathology and Microbiology,1 and Department of Pathology and Microbiology,2 University of Nebraska Medical Center, Omaha, Nebraska 68198

Received 17 December 1999/Accepted 8 June 2000

We cloned the sequence encoding murine interleukin-4 (mIL-4), including the secretory signal, into the genome of CVB3/0, an artificially attenuated strain of coxsackievirus B3, at the junction of the capsid protein 1D and the viral protease 2Apro. Two strains of chimeric CVB3 were constructed using, in one case, identical sequences to encode 2Apro cleavage sites (CVB3/0-mIL4/47) on either side of the inserted coding sequence and, in the other case, nonidentical sequences that varied at the nucleotide level without changing the amino acid sequences (CVB3-PL2-mIL4/46). Transfection of HeLa cells yielded progeny viruses that replicated with rates similar to that of the parental CVB3/0 strain, although yields of mIL-4-expressing strains were approximately 10-fold lower than those of the parental virus. Western blot analysis of viral proteins isolated from HeLa cells inoculated with either strain of chimeric virus demonstrated that the chimeric viruses synthesized capsid protein 1D at approximately twofold-higher levels than the parental virus. mIL-4 protein was detected by enzyme-linked immunosorbent assay (ELISA) in HeLa cells inoculated with either strain of chimeric virus. Lysates of HeLa cells inoculated with either chimeric virus induced the proliferation of the mIL-4-requiring murine MC-9 cell line, demonstrating biological activity of the CVB3-expressed mIL-4. Reverse transcription (RT)-PCR analysis of viral RNA derived from sequential passaging of CVB3/0-mIL4/47 in HeLa cells demonstrated deletion of the mIL-4 coding sequence occurring by the fourth passage, while similar analysis of CVB3-PL2-mIL4/46 RNA demonstrated detection of the mIL-4 coding sequence in the virus population through 10 generations in HeLa cells. mIL-4 protein levels determined by ELISA were consistent with the stability and loss data determined by RT-PCR analysis of the passaged viral genomes. Studies of insert stability of CVB3-PL2-mIL4/46 during replication in mice showed the presence of the viral mIL-4 insert in pancreas, heart, and liver at 14 days postinfection. Comparison of the murine antibody responses to CVB3-PL2-mIL4/46 and the parental CVB3/0 strain demonstrated an increased level of CVB3-binding serum immunoglobulin G1 in mice inoculated with CVB3-PL2-mIL4/46.

* Corresponding author. Mailing address: Enterovirus Research Laboratory, Department of Pathology and Microbiology, University of Nebraska Medical Center, 986495 Nebraska Medical Center, Omaha, NE 68198. Phone: (402) 559-7735. Fax: (402) 559-4077. E-mail: nchapman{at}unmc.edu.

dagger Present address: Transfusion Service, Stanford University Hospital, Stanford, CA 94303.

Dagger Present address: Path Logic, Inc., Dixon, CA 95620.

Journal of Virology, September 2000, p. 7952-7962, Vol. 74, No. 17
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Yajima, T., Knowlton, K. U. (2009). Viral Myocarditis: From the Perspective of the Virus. Circulation 119: 2615-2624 [Full Text]  
  • Carson, S. D., Kim, K.-S., Pirruccello, S. J., Tracy, S., Chapman, N. M. (2007). Endogenous low-level expression of the coxsackievirus and adenovirus receptor enables coxsackievirus B3 infection of RD cells. J. Gen. Virol. 88: 3031-3038 [Abstract] [Full Text]  
  • Jerrells, T. R., Vidlak, D., Strachota, J. M. (2007). Alcoholic pancreatitis: mechanisms of viral infections as cofactors in the development of acute and chronic pancreatitis and fibrosis. J. Leukoc. Biol. 81: 430-439 [Abstract] [Full Text]  
  • Dan, M., Chantler, J. K. (2005). A Genetically Engineered Attenuated Coxsackievirus B3 Strain Protects Mice against Lethal Infection. J. Virol. 79: 9285-9295 [Abstract] [Full Text]  
  • Kim, K.-S., Tracy, S., Tapprich, W., Bailey, J., Lee, C.-K., Kim, K., Barry, W. H., Chapman, N. M. (2005). 5'-Terminal Deletions Occur in Coxsackievirus B3 during Replication in Murine Hearts and Cardiac Myocyte Cultures and Correlate with Encapsidation of Negative-Strand Viral RNA. J. Virol. 79: 7024-7041 [Abstract] [Full Text]  
  • Lee, C.-K., Kono, K., Haas, E., Kim, K.-S., Drescher, K. M., Chapman, N. M., Tracy, S. (2005). Characterization of an infectious cDNA copy of the genome of a naturally occurring, avirulent coxsackievirus B3 clinical isolate. J. Gen. Virol. 86: 197-210 [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]  
  • Henke, A., Zell, R., Ehrlich, G., Stelzner, A. (2001). Expression of Immunoregulatory Cytokines by Recombinant Coxsackievirus B3 Variants Confers Protection against Virus-Caused Myocarditis. J. Virol. 75: 8187-8194 [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»