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Human cytomegalovirus protein pp65: an efficient protein carrier system into human dendritic cells

Gene Therapy volume 15, pages 318–325 (2008)Cite this article

Abstract

Protein-based immunogens are usually poor inducers of CD8+ T cells. To enhance the induction of CD8+ T cells, one approach is the use of protein immunogens coupled to protein transduction domains (PTDs). These are small cationic peptide sequences that significantly enhance the uptake of fused proteins into dendritic cells (DC) and then mediate their presentation in the context of major histocompatibility complex class I (MHC-I) and MHC-II molecules. One drawback of this system is the high concentrations of PTD-fusion proteins required. Here, we show that proteins fused to the human cytomegalovirus tegument protein pp65 were bound with higher efficiency to DCs than those fused to the described PTDs TatPTD and Penetratin. Furthermore, the fusion of pp65 to proteins led to an enhanced uptake of these proteins by DCs. Once taken up, CD4+ and CD8+ memory T cells were strongly stimulated ex vivo demonstrating that pp65 was efficiently processed and presented in the context of both MHC-I and MHC-II. These data make pp65 a promising delivery system to induce cellular immune responses by fused protein vaccines.

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Acknowledgements

This work was supported by the Saarland University. We thank T Tänzer and D Holzer for technical assistance. We thank the Winterbergkliniken Saarbrücken for providing us with blood cell concentrates and additional information about the HCMV serostatus of the blood donors. We thank, furthermore, the volunteers for the donation of their blood. We also thank Frank T Hufert for a plasmid containing pp65 cDNA.

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Author notes

  1. R Furtwängler

    Present address: 4Current address: Department of Pediatric Hematology and Oncology, Saarland University, 66421 Homburg, Germany.,

Authors and Affiliations

  1. Department of Virology, Saarland University, Homburg, Germany

    N Scheller, R Furtwängler, T Breinig & A Meyerhans

  2. Medical Department IV, Saarland University, Homburg, Germany

    U Sester

  3. Research Department, Kantonal Hospital St Gallen, St Gallen, Switzerland

    R Maier

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Correspondence to A Meyerhans.

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Supplementary Information accompanies the paper on Gene Therapy website (http://www.nature.com/gt)

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Scheller, N., Furtwängler, R., Sester, U. et al. Human cytomegalovirus protein pp65: an efficient protein carrier system into human dendritic cells. Gene Ther 15, 318–325 (2008). https://doi.org/10.1038/sj.gt.3303086

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