核糖核酸
翻译效率
生物
信使核糖核酸
抄写(语言学)
非翻译区
寡核苷酸
抗原
化学
分子生物学
细胞生物学
转染
DNA
基因
翻译(生物学)
生物化学
遗传学
语言学
哲学
作者
Silke Holtkamp,Sebastian Kreiter,Abderraouf Selmi,Petra Simon,Michael Koslowski,Christoph Huber,Özlem Türeci,Uğur Şahin
出处
期刊:Blood
[American Society of Hematology]
日期:2006-12-15
卷期号:108 (13): 4009-4017
被引量:442
标识
DOI:10.1182/blood-2006-04-015024
摘要
Abstract 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 β-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.
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