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Depletion of CD25+ cells from human T-cell enriched fraction eliminates immunodominance during priming with dendritic cells genetically modified to express a secreted protein

Cancer Gene Therapy volume 12, pages 185–197 (2005)Cite this article

Abstract

The ability of dendritic cells (DCs), genetically modified with one of two types of plasmid DNA vaccines to stimulate lymphocytes from normal human donors and to generate antigen-specific responses, is compared. The first type, also called “secreted” vaccine (sVac), encodes for the full length of the human prostate-specific antigen (PSA) with a signal peptide sequence so that the expressed product is glycosylated and directed to the secretory pathway. The second type, truncated vaccines (tVacs), encodes for either hPSA or human prostate acidic phosphatase (hPAP), both of which lack signal peptide sequences and are retained in the cytosol and degraded by the proteasomes following expression. Monocyte-derived dendritic cells are transiently transfected with either sVac or one of two tVacs. The DCs are then used to activate CD25+-depleted or nondepleted autologous lymphocytes in an in vitro model of DNA vaccination. Lymphocytes are boosted following priming with transfected DCs, peptide-pulsed DCs or monocytes. Their reactivity is tested against tumor cells or peptide-pulsed T2 target cells. Both tVacDCs and sVacDCs generate antigen-specific cytotoxic T-cell responses. The immune response is restricted towards one of the three antigen-derived epitopes when priming and boosting is performed with sVacDCs. In contrast, tVac-transfected DCs prime T cells towards all antigen-derived epitopes. Subsequent repeated boosting with transfected DCs, however, restricts the immune response to a single epitope due to immunodominance. While CD25+ cell depletion prior to priming with sVacDCs alleviates immunodominance, cotransfection of dendritic cells with GITR-L does so in some but not all cases.

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Acknowledgements

This work was supported in part by Grant N00014-00-1-0787 from the Office of Naval Research, and award No DAMD17-02-1-0239. The US Army Medical Research Acquisition Activity, 820 Chandler Street, Fort Detrick, MD 21702-5014 is the awarding and administering acquisition office.

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Authors and Affiliations

  1. The George Washington University Medical Center, Washington, DC, USA

    Milcho Mincheff, Serguei Zoubak, Petro Pogribnyy, Yevgen Makogonenko & Harold T Meryman

  2. Central Laboratory for Immunology, St Ivan Rilski Hospital, Sofia, Bulgaria

    Iskra Altankova

  3. Department of Urology, St Ann University Hospital, Sofia, Bulgaria

    Stoyan Tchakarov

  4. National Institute of Hematology and Transfusion Medicine, Sofia, Bulgaria

    Milcho Mincheff & Chavdar Botev

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  1. Milcho Mincheff
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Correspondence to Milcho Mincheff.

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Mincheff, M., Zoubak, S., Altankova, I. et al. Depletion of CD25+ cells from human T-cell enriched fraction eliminates immunodominance during priming with dendritic cells genetically modified to express a secreted protein. Cancer Gene Ther 12, 185–197 (2005). https://doi.org/10.1038/sj.cgt.7700778

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