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Human dendritic cells genetically engineered to express cytosolically retained fragment of prostate-specific membrane antigen prime cytotoxic T-cell responses to multiple epitopes

Cancer Gene Therapy volume 10, pages 907–917 (2003)Cite this article

Abstract

The ability of two plasmid DNA vaccines to stimulate lymphocytes from normal human donors and to generate antigen-specific responses is demonstrated. The first vaccine (truncated; tPSMA) encodes for only the extracellular domain of prostate-specific membrane antigen (PSMA). The product, expressed following transfection with this vector, is retained in the cytosol and degraded by the proteasomes. For the “secreted” (sPMSA) vaccine, a signal peptide sequence is added to the expression cassette and the expressed protein is glycosylated and directed to the secretory pathway. Monocyte-derived dendritic cells (DCs) are transiently transfected with either sPSMA or tPSMA plasmids. The DCs are then used to activate autologous lymphocytes in an in vitro model of DNA vaccination. Lymphocytes are boosted following priming with transfected DCs or with peptide-pulsed monocytes. Their reactivity is tested against tumor cells or peptide-pulsed T2 target cells. Both tPSMA DCs and sPSMA DCs generate antigen-specific cytotoxic T-cell responses. The immune response is restricted toward one of the four PSMA-derived epitopes when priming and boosting is performed with sPSMA. In contrast, tPSMA-transfected DCs prime T cells toward several PSMA-derived epitopes. Subsequent repeated boosting with transfected DCs, however, restricts the immune response to a single epitope due to immunodominance.

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Acknowledgements

This work was supported in part by Grant N00014-00-1-0787 from the Office of Naval Research and by 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. The content of the information does not necessarily reflect the position or the policy of the Government, and no official endorsement should be inferred. For the purpose of this article, information includes news releases, articles, manuscripts, brochures, advertisements, still and motion pictures, speeches, trade association proceedings, etc.

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Authors and Affiliations

  1. The George Washington University Medical Center, Washington, District Columbia, USA

    Milcho Mincheff, Serguei Zoubak, Yevgen Makogonenko, Michael Hammett & Harold Meryman

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

    Iskra Altankova & Yavor Pomakov

  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, Irena Ignatova, Rosen Dimitrov, Kalina Madarzhieva & Toshko Lissitchkov

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Correspondence to Milcho Mincheff.

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Mincheff, M., Zoubak, S., Altankova, I. et al. Human dendritic cells genetically engineered to express cytosolically retained fragment of prostate-specific membrane antigen prime cytotoxic T-cell responses to multiple epitopes. Cancer Gene Ther 10, 907–917 (2003). https://doi.org/10.1038/sj.cgt.7700647

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