SEARCH:   Advanced

Blood, 15 December 2006, Vol. 108, No. 13, pp. 4009-4017. Prepublished online as a Blood First Edition Paper on August 29, 2006; DOI 10.1182/blood-2006-04-015024. This Article Full Text Full Text (PDF) All Versions of this Article: blood-2006-04-015024v1 108/13/4009    most recent Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Rights and Permissions Citing Articles Citing Articles via HighWire Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by Holtkamp, S. Articles by Sahin, U. Search for Related Content PubMed PubMed Citation Articles by Holtkamp, S. Articles by Sahin, U. Related Collections Immunobiology Gene Therapy Social Bookmarking                   What's this?  Previous Article  |  Table of Contents  |  Next Article  GENE THERAPY Modification of antigen-encoding RNA increases stability, translational efficacy, and T-cell stimulatory capacity of dendritic cells Silke Holtkamp, Sebastian Kreiter, Abderraouf Selmi, Petra Simon, Michael Koslowski, Christoph Huber, Özlem Türeci, and Ugur Sahin From the Department of Internal Medicine III, Division of Experimental and Translational Oncology, Johannes-Gutenberg University, Mainz, Germany. Adoptive transfer of dendritic cells (DCs) transfected with in vitro–transcribed, RNA-encoding, tumor-associated antigens has recently entered clinical testing as a promising approach for cancer immunotherapy. However, pharmacokinetic exploration of RNA as a potential drug compound and a key aspect of clinical development is still pending. While investigating the impact of different structural modifications of RNA molecules on the kinetics of the encoded protein in DCs, we identified components located 3' of the coding region that contributed to a higher transcript stability and translational efficiency. With the use of quantitative reverse transcription–polymerase chain reaction (RT-PCR) and eGFP variants to measure transcript amounts and protein yield, we showed that a poly(A) tail measuring 120 nucleotides compared with a shorter one, an unmasked poly(A) tail with a free 3' end rather than one extended with unrelated nucleotides, and 2 sequential -globin 3' untranslated regions cloned head to tail between the coding region and the poly(A) tail each independently enhanced RNA stability and translational efficiency. Consecutively, the density of antigen-specific peptide/MHC complexes on the transfected cells and their potency to stimulate and expand antigen-specific CD4+ and CD8+ T cells were also increased. In summary, our data provide a strategy for optimizing RNA-transfected DC vaccines and a basis for defining release criteria for such vaccine preparations. CiteULike    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this? This article has been cited by other articles: M. Koslowski, O. Tureci, S. Biesterfeld, G. Seitz, C. Huber, and U. Sahin Selective Activation of Trophoblast-specific PLAC1 in Breast Cancer by CCAAT/Enhancer-binding Protein {beta} (C/EBP{beta}) Isoform 2 J. Biol. Chem., October 16, 2009; 284(42): 28607 - 28615. [Abstract] [Full Text] [PDF] S.-i. Fujii, A. Goto, and K. Shimizu Antigen mRNA-transfected, allogeneic fibroblasts loaded with NKT-cell ligand confer antitumor immunity Blood, April 30, 2009; 113(18): 4262 - 4272. [Abstract] [Full Text] [PDF] S. Kreiter, A. Selmi, M. Diken, M. Sebastian, P. Osterloh, H. Schild, C. Huber, O. Tureci, and U. Sahin Increased Antigen Presentation Efficiency by Coupling Antigens to MHC Class I Trafficking Signals J. Immunol., January 1, 2008; 180(1): 309 - 318. [Abstract] [Full Text] [PDF]

	Copyright © 2006 by American Society of Hematology         Online ISSN: 1528-0020