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 Rodriguez, F.
Right arrow Articles by Whitton, J. L.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Rodriguez, F.
Right arrow Articles by Whitton, J. L.

 Previous Article  |  Next Article 

Journal of Virology, November 2001, p. 10421-10430, Vol. 75, No. 21
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.21.10421-10430.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

CD4+ T Cells Induced by a DNA Vaccine: Immunological Consequences of Epitope-Specific Lysosomal Targeting†

Fernando Rodriguez,1,2 Stephanie Harkins,1 Jeffrey M. Redwine,1 Jose M. de Pereda,3 and J. Lindsay Whitton1,*

Department of Neuropharmacology, The Scripps Research Institute,1 and The Burnham Institute,3 La Jolla, California, and Unidad de Investigación, Hospital Universitario 12 de Octubre, Madrid, Spain2

Received 5 April 2001/Accepted 19 July 2001

Our previous studies have shown that targeting DNA vaccine-encoded major histocompatibility complex class I epitopes to the proteasome enhanced CD8+ T-cell induction and protection against lymphocytic choriomeningitis virus (LCMV) challenge. Here, we expand these studies to evaluate CD4+ T-cell responses induced by DNA immunization and describe a system for targeting proteins and minigenes to lysosomes. Full-length proteins can be targeted to the lysosomal compartment by covalent attachment to the 20-amino-acid C-terminal tail of lysosomal integral membrane protein-II (LIMP-II). Using minigenes encoding defined T-helper epitopes from lymphocytic choriomeningitis virus, we show that the CD4+ T-cell response induced by the NP309-328 epitope of LCMV was greatly enhanced by addition of the LIMP-II tail. However, the immunological consequence of lysosomal targeting is not invariably positive; the CD4+ T-cell response induced by the GP61-80 epitope was almost abolished when attached to the LIMP-II tail. We identify the mechanism which underlies this marked difference in outcome. The GP61-80 epitope is highly susceptible to cleavage by cathepsin D, an aspartic endopeptidase found almost exclusively in lysosomes. We show, using mass spectrometry, that the GP61-80 peptide is cleaved between residues F74 and K75 and that this destroys its ability to stimulate virus-specific CD4+ T cells. Thus, the immunological result of lysosomal targeting varies, depending upon the primary sequence of the encoded antigen. We analyze the effects of CD4+ T-cell priming on the virus-specific antibody and CD8+ T-cell responses which are mounted after virus infection and show that neither response appears to be accelerated or enhanced. Finally, we evaluate the protective benefits of CD4+ T-cell vaccination in the LCMV model system; in contrast to DNA vaccine-induced CD8+ T cells, which can confer solid protection against LCMV challenge, DNA vaccine-mediated priming of CD4+ T cells does not appear to enhance the vaccinee's ability to combat viral challenge.

* 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 This is manuscript number 13917-NP from The Scripps Research Institute.

Journal of Virology, November 2001, p. 10421-10430, Vol. 75, No. 21
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.21.10421-10430.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Takagi, A., Matsui, M., Ohno, S., Duan, H., Moriya, O., Kobayashi, N., Oda, H., Mori, M., Kobayashi, A., Taneichi, M., Uchida, T., Akatsuka, T. (2009). Highly Efficient Antiviral CD8+ T-Cell Induction by Peptides Coupled to the Surfaces of Liposomes. CVI 16: 1383-1392 [Abstract] [Full Text]  
  • Fernandez-Borges, N., Brun, A., Whitton, J. L., Parra, B., Diaz-San Segundo, F., Salguero, F. J., Torres, J. M., Rodriguez, F. (2006). DNA Vaccination Can Break Immunological Tolerance to PrP in Wild-Type Mice and Attenuates Prion Disease after Intracerebral Challenge.. J. Virol. 80: 9970-9976 [Abstract] [Full Text]  
  • Arruda, L. B., Sim, D., Chikhlikar, P. R., Maciel, M. Jr, Akasaki, K., August, J. T., Marques, E. T. A. (2006). Dendritic Cell-Lysosomal-Associated Membrane Protein (LAMP) and LAMP-1-HIV-1 Gag Chimeras Have Distinct Cellular Trafficking Pathways and Prime T and B Cell Responses to a Diverse Repertoire of Epitopes. J. Immunol. 177: 2265-2275 [Abstract] [Full Text]  
  • Maksimow, M., Miiluniemi, M., Marttila-Ichihara, F., Jalkanen, S., Hanninen, A. (2006). Antigen targeting to endosomal pathway in dendritic cell vaccination activates regulatory T cells and attenuates tumor immunity. Blood 108: 1298-1305 [Abstract] [Full Text]  
  • Ko, H.-J., Ko, S.-Y., Kim, Y.-J., Lee, E.-G., Cho, S.-N., Kang, C.-Y. (2005). Optimization of Codon Usage Enhances the Immunogenicity of a DNA Vaccine Encoding Mycobacterial Antigen Ag85B. Infect. Immun. 73: 5666-5674 [Abstract] [Full Text]  
  • Marques, E. T. A. Jr., Chikhlikar, P., de Arruda, L. B., Leao, I. C., Lu, Y., Wong, J., Chen, J.-S., Byrne, B., August, J. T. (2003). HIV-1 p55Gag Encoded in the Lysosome-associated Membrane Protein-1 as a DNA Plasmid Vaccine Chimera Is Highly Expressed, Traffics to the Major Histocompatibility Class II Compartment, and Elicits Enhanced Immune Responses. J. Biol. Chem. 278: 37926-37936 [Abstract] [Full Text]  
  • Rush, C., Mitchell, T., Garside, P. (2002). Efficient Priming of CD4+ and CD8+ T Cells by DNA Vaccination Depends on Appropriate Targeting of Sufficient Levels of Immunologically Relevant Antigen to Appropriate Processing Pathways. J. Immunol. 169: 4951-4960 [Abstract] [Full Text]  
  • Livingston, B., Crimi, C., Newman, M., Higashimoto, Y., Appella, E., Sidney, J., Sette, A. (2002). A Rational Strategy to Design Multiepitope Immunogens Based on Multiple Th Lymphocyte Epitopes. J. Immunol. 168: 5499-5506 [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»