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The cationic region from HIV tat enhances the cell-surface expression of epitope/MHC class I complexes

Gene Therapy volume 10, pages 2067–2073 (2003)Cite this article

Abstract

The potential of genetic immunization has been acknowledged for almost a decade, but disappointing immunogenicity in humans has delayed its introduction into the clinical arena. To try to increase the potency of genetic immunization, we and others have evaluated ‘translocatory’ proteins, which are thought to exit living cells by an uncharacterized pathway, and enter neighboring cells in an energy-independent manner. Several laboratories, including our own, have begun to question these remarkable properties. Our previous studies showed that the ability of an epitope to induce major histocompatibility complex (MHC) class I restricted CD8+ T cells was, indeed, enhanced by its being attached to the proposed translocatory sequence of the HIV-1 tat protein. However, we found little evidence that the increased immunogenicity resulted from transfer of the fusion peptide between living cells, and we proposed that it resulted instead from an increased epitope/MHC expression on the surface of transfected cells. Here, we directly test this hypothesis. We show that cells cotransfected with plasmids encoding an epitope, and the relevant MHC class I allele, can stimulate epitope-specific T cells, and that attachment of the epitope to a putative translocatory sequence – which we term herein an ‘integral cationic region’ (ICR) – leads to a marked increase in stimulatory activity. This elevated stimulatory capacity does not result from a nonspecific increase in MHC class I expression. We use a high-affinity T-cell receptor (TcR) specific for the epitope/MHC combination to quantitate directly the cell-surface expression of the immunogenic complex, and we show that the attachment of the tat ICR to an epitope results in a substantial enhancement of its cell-surface presentation. These data suggest an alternative explanation for the immune enhancement seen with ICRs.

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Acknowledgements

We are grateful to Annette Lord for excellent secretarial support. This work was supported by NIH Grants AI-37186 (to JLW), GM55767 (to DMK), and D/99/22463 from the Deutsche Krebshilfe e.V. (to JAL). This is manuscript number 15238-NP from the Scripps Research Institute.

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Authors and Affiliations

  1. Department of Neuropharmacology, The Scripps Research Institute, La Jolla, CA, USA

    J A Leifert, S Harkins & J L Whitton

  2. Department of Biochemistry, University of Illinois, Urbana, IL, USA

    P D Holler & D M Kranz

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  1. J A Leifert
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Leifert, J., Holler, P., Harkins, S. et al. The cationic region from HIV tat enhances the cell-surface expression of epitope/MHC class I complexes. Gene Ther 10, 2067–2073 (2003). https://doi.org/10.1038/sj.gt.3302115

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