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 Smerdou, C.
Right arrow Articles by Liljeström, P.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Smerdou, C.
Right arrow Articles by Liljeström, P.

 Previous Article  |  Next Article 

Journal of Virology, February 1999, p. 1092-1098, Vol. 73, No. 2
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Two-Helper RNA System for Production of Recombinant Semliki Forest Virus Particles

C. Smerdou1 and P. Liljeström1,2,*

Microbiology and Tumor Biology Center, Karolinska Institute, S-17177 Stockholm,1 and Department of Vaccine Research, Swedish Institute for Infectious Disease Control, S-17182 Solna,2 Sweden

Received 18 September 1998/Accepted 30 October 1998

Alphavirus expression systems based on suicidal virus particles carrying recombinant replicons have proven to be a very efficient way to deliver genes for heterologous protein expression. However, present strategies for production of such particles have biosafety limitations due to the generation, by RNA recombination, of replication-proficient viruses (RPVs). Here we describe a new packaging system for Semliki Forest virus (SFV) based on a the use of a two-helper system in which the capsid and spike proteins of the C-p62-6K-E1 polyprotein are expressed from two independent RNA molecules. The capsid gene contains a translational enhancer and therefore that sequence was also engineered in front of the spike sequence p62-6K-E1. A sequence coding for the foot-and-mouth disease virus 2A autoprotease was inserted in frame between the capsid translational enhancer and the spike genes. This allows production of the spike proteins at high levels with cotranslational removal of the enhancer sequence and normal biosynthesis of the spike complex. The autoprotease activity of the capsid protein was abolished by mutation, further increasing the biosafety of the system. Cotransfection of cells with both helper RNAs and an SFV vector replicon carrying the LacZ gene led to production of recombinant particles with titers of up to 8 × 108 particles per 106 cells. Extensive analysis failed to demonstrate the presence of any RPVs, emphasizing the high biosafety of the system based on two-helper RNAs.

* Corresponding author. Mailing address: Microbiology and Tumor Biology Center, Karolinska Institute, Box 280, S-17177 Stockholm, Sweden. Phone: 46-8-457 2550. Fax: 46-8-310848. E-mail: peter.liljestrom{at}mtc.ki.se.

Journal of Virology, February 1999, p. 1092-1098, Vol. 73, No. 2
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Rodriguez-Madoz, J. R., Liu, K. H., Quetglas, J. I., Ruiz-Guillen, M., Otano, I., Crettaz, J., Butler, S. D., Bellezza, C. A., Dykes, N. L., Tennant, B. C., Prieto, J., Gonzalez-Aseguinolaza, G., Smerdou, C., Menne, S. (2009). Semliki Forest Virus Expressing Interleukin-12 Induces Antiviral and Antitumoral Responses in Woodchucks with Chronic Viral Hepatitis and Hepatocellular Carcinoma. J. Virol. 83: 12266-12278 [Abstract] [Full Text]  
  • Meseda, C. A., Mayer, A. E., Kumar, A., Garcia, A. D., Campbell, J., Listrani, P., Manischewitz, J., King, L. R., Golding, H., Merchlinsky, M., Weir, J. P. (2009). Comparative Evaluation of the Immune Responses and Protection Engendered by LC16m8 and Dryvax Smallpox Vaccines in a Mouse Model. CVI 16: 1261-1271 [Abstract] [Full Text]  
  • Mooij, P., Balla-Jhagjhoorsingh, S. S., Beenhakker, N., van Haaften, P., Baak, I., Nieuwenhuis, I. G., Heidari, S., Wolf, H., Frachette, M.-J., Bieler, K., Sheppard, N., Harari, A., Bart, P.-A., Liljestrom, P., Wagner, R., Pantaleo, G., Heeney, J. L. (2009). Comparison of Human and Rhesus Macaque T-Cell Responses Elicited by Boosting with NYVAC Encoding Human Immunodeficiency Virus Type 1 Clade C Immunogens. J. Virol. 83: 5881-5889 [Abstract] [Full Text]  
  • Chan, C.-M., Woo, P. C. Y., Lau, S. K. P., Tse, H., Chen, H.-L., Li, F., Zheng, B.-J., Chen, L., Huang, J.-D., Yuen, K.-Y. (2008). Spike Protein, S, of Human Coronavirus HKU1: Role in Viral Life Cycle and Application in Antibody Detection. Exp. Biol. Med. 233: 1527-1536 [Abstract] [Full Text]  
  • Tamm, K., Merits, A., Sarand, I. (2008). Mutations in the nuclear localization signal of nsP2 influencing RNA synthesis, protein expression and cytotoxicity of Semliki Forest virus. J. Gen. Virol. 89: 676-686 [Abstract] [Full Text]  
  • Fragkoudis, R., Breakwell, L., McKimmie, C., Boyd, A., Barry, G., Kohl, A., Merits, A., Fazakerley, J. K. (2007). The type I interferon system protects mice from Semliki Forest virus by preventing widespread virus dissemination in extraneural tissues, but does not mediate the restricted replication of avirulent virus in central nervous system neurons. J. Gen. Virol. 88: 3373-3384 [Abstract] [Full Text]  
  • Forsell, M. N. E., McInerney, G. M., Dosenovic, P., Hidmark, A. S., Eriksson, C., Liljestrom, P., Grundner, C., Karlsson Hedestam, G. B. (2007). Increased human immunodeficiency virus type 1 Env expression and antibody induction using an enhanced alphavirus vector. J. Gen. Virol. 88: 2774-2779 [Abstract] [Full Text]  
  • Garcia, A. D., Meseda, C. A., Mayer, A. E., Kumar, A., Merchlinsky, M., Weir, J. P. (2007). Characterization and Use of Mammalian-Expressed Vaccinia Virus Extracellular Membrane Proteins for Quantification of the Humoral Immune Response to Smallpox Vaccines. CVI 14: 1032-1044 [Abstract] [Full Text]  
  • Berglund, P., Finzi, D., Bennink, J. R., Yewdell, J. W. (2007). Viral Alteration of Cellular Translational Machinery Increases Defective Ribosomal Products. J. Virol. 81: 7220-7229 [Abstract] [Full Text]  
  • Naslund, T. I., Uyttenhove, C., Nordstrom, E. K. L., Colau, D., Warnier, G., Jondal, M., Van den Eynde, B. J., Liljestrom, P. (2007). Comparative Prime-Boost Vaccinations Using Semliki Forest Virus, Adenovirus, and ALVAC Vectors Demonstrate Differences in the Generation of a Protective Central Memory CTL Response against the P815 Tumor. J. Immunol. 178: 6761-6769 [Abstract] [Full Text]  
  • Douagi, I., McInerney, G. M., Hidmark, A. S., Miriallis, V., Johansen, K., Svensson, L., Karlsson Hedestam, G. B. (2007). Role of Interferon Regulatory Factor 3 in Type I Interferon Responses in Rotavirus-Infected Dendritic Cells and Fibroblasts. J. Virol. 81: 2758-2768 [Abstract] [Full Text]  
  • Hidmark, A. S., Nordstrom, E. K. L., Dosenovic, P., Forsell, M. N. E., Liljestrom, P., Karlsson Hedestam, G. B. (2006). Humoral Responses against Coimmunized Protein Antigen but Not against Alphavirus-Encoded Antigens Require Alpha/Beta Interferon Signaling. J. Virol. 80: 7100-7110 [Abstract] [Full Text]  
  • Guan, M., Rodriguez-Madoz, J. R., Alzuguren, P., Gomar, C., Kramer, M. G., Kochanek, S., Prieto, J., Smerdou, C., Qian, C. (2006). Increased Efficacy and Safety in the Treatment of Experimental Liver Cancer with a Novel Adenovirus-Alphavirus Hybrid Vector. Cancer Res. 66: 1620-1629 [Abstract] [Full Text]  
  • Diatta, A., Piver, E., Collin, C., Vaudin, P., Pages, J.-C. (2005). Semliki Forest virus-derived virus-like particles: characterization of their production and transduction pathways. J. Gen. Virol. 86: 3129-3136 [Abstract] [Full Text]  
  • Forsell, M. N. E., Li, Y., Sundback, M., Svehla, K., Liljestrom, P., Mascola, J. R., Wyatt, R., Hedestam, G. B. K. (2005). Biochemical and Immunogenic Characterization of Soluble Human Immunodeficiency Virus Type 1 Envelope Glycoprotein Trimers Expressed by Semliki Forest Virus. J. Virol. 79: 10902-10914 [Abstract] [Full Text]  
  • Hidmark, A. S., McInerney, G. M., Nordstrom, E. K. L., Douagi, I., Werner, K. M., Liljestrom, P., Hedestam, G. B. K. (2005). Early Alpha/Beta Interferon Production by Myeloid Dendritic Cells in Response to UV-Inactivated Virus Requires Viral Entry and Interferon Regulatory Factor 3 but Not MyD88. J. Virol. 79: 10376-10385 [Abstract] [Full Text]  
  • McInerney, G. M., Kedersha, N. L., Kaufman, R. J., Anderson, P., Liljestrom, P. (2005). Importance of eIF2{alpha} Phosphorylation and Stress Granule Assembly in Alphavirus Translation Regulation. Mol. Biol. Cell 16: 3753-3763 [Abstract] [Full Text]  
  • Garcia, S., Billecocq, A., Crance, J.-M., Munderloh, U., Garin, D., Bouloy, M. (2005). Nairovirus RNA Sequences Expressed by a Semliki Forest Virus Replicon Induce RNA Interference in Tick Cells. J. Virol. 79: 8942-8947 [Abstract] [Full Text]  
  • Onate, A. A., Donoso, G., Moraga-Cid, G., Folch, H., Cespedes, S., Andrews, E. (2005). An RNA Vaccine Based on Recombinant Semliki Forest Virus Particles Expressing the Cu,Zn Superoxide Dismutase Protein of Brucella abortus Induces Protective Immunity in BALB/c Mice. Infect. Immun. 73: 3294-3300 [Abstract] [Full Text]  
  • Chen, M., Barnfield, C., Naslund, T. I., Fleeton, M. N., Liljestrom, P. (2005). MyD88 Expression Is Required for Efficient Cross-Presentation of Viral Antigens from Infected Cells. J. Virol. 79: 2964-2972 [Abstract] [Full Text]  
  • Rheme, C., Ehrengruber, M. U., Grandgirard, D. (2005). Alphaviral cytotoxicity and its implication in vector development. Exp Physiol 90: 45-52 [Abstract] [Full Text]  
  • Gehrke, R., Ecker, M., Aberle, S. W., Allison, S. L., Heinz, F. X., Mandl, C. W. (2003). Incorporation of Tick-Borne Encephalitis Virus Replicons into Virus-Like Particles by a Packaging Cell Line. J. Virol. 77: 8924-8933 [Abstract] [Full Text]  
  • Thomas, J. M., Klimstra, W. B., Ryman, K. D., Heidner, H. W. (2003). Sindbis Virus Vectors Designed To Express a Foreign Protein as a Cleavable Component of the Viral Structural Polyprotein. J. Virol. 77: 5598-5606 [Abstract] [Full Text]  
  • Hanke, T., Barnfield, C., Wee, E. G.-T., Agren, L., Samuel, R. V., Larke, N., Liljestrom, P. (2003). Construction and immunogenicity in a prime-boost regimen of a Semliki Forest virus-vectored experimental HIV clade A vaccine. J. Gen. Virol. 84: 361-368 [Abstract] [Full Text]  
  • Palmowski, M. J., Choi, E. M.-L., Hermans, I. F., Gilbert, S. C., Chen, J.-L., Gileadi, U., Salio, M., Van Pel, A., Man, S., Bonin, E., Liljestrom, P., Dunbar, P. R., Cerundolo, V. (2002). Competition Between CTL Narrows the Immune Response Induced by Prime-Boost Vaccination Protocols. J. Immunol. 168: 4391-4398 [Abstract] [Full Text]  
  • Brinster, C., Chen, M., Boucreux, D., Paranhos-Baccala, G., Liljestrom, P., Lemmonier, F., Inchauspe, G. (2002). Hepatitis C virus non-structural protein 3-specific cellular immune responses following single or combined immunization with DNA or recombinant Semliki Forest virus particles. J. Gen. Virol. 83: 369-381 [Abstract] [Full Text]  
  • Donnelly, M. L. L., Hughes, L. E., Luke, G., Mendoza, H., ten Dam, E., Gani, D., Ryan, M. D. (2001). The 'cleavage' activities of foot-and-mouth disease virus 2A site-directed mutants and naturally occurring '2A-like' sequences. J. Gen. Virol. 82: 1027-1041 [Abstract] [Full Text]  
  • Pugachev, K. V., Tzeng, W.-P., Frey, T. K. (2000). Development of a Rubella Virus Vaccine Expression Vector: Use of a Picornavirus Internal Ribosome Entry Site Increases Stability of Expression. J. Virol. 74: 10811-10815 [Abstract] [Full Text]  
  • Heise, M. T., Simpson, D. A., Johnston, R. E. (2000). Sindbis-Group Alphavirus Replication in Periosteum and Endosteum of Long Bones in Adult Mice. J. Virol. 74: 9294-9299 [Abstract] [Full Text]  
  • Fleeton, M. N., Liljeström, P., Sheahan, B. J., Atkins, G. J. (2000). Recombinant Semliki Forest virus particles expressing louping ill virus antigens induce a better protective response than plasmid-based DNA vaccines or an inactivated whole particle vaccine. J. Gen. Virol. 81: 749-758 [Abstract] [Full Text]  
  • Atkins, G. J., Sheahan, B. J., Liljeström, P. (1999). The molecular pathogenesis of Semliki Forest virus: a model virus made useful?. J. Gen. Virol. 80: 2287-2297 [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»