Jonuleit H, Tüting T, Steitz J, Brück J, Giesecke A, Steinbrink K, Knop J, Enk A H
Department of Dermatology, J Gutenberg-University, Langenbeckstrasse 1, D-55101 Mainz, Germany.
Gene Ther. 2000 Feb;7(3):249-54. doi: 10.1038/sj.gt.3301077.
We have developed a culture method for the foreign serum-free generation of highly immunostimulatory, CD83+ human dendritic cells (DC). In this study, we evaluated the feasibility and consequences of endogenously expressing antigens in mature DC using adenoviral vectors. Transduction of DC with Ad-EGFP demonstrated endogenous fluorescence in 50-85% of CD83+ DC. Ad-transduced DC stimulated the proliferation of allogeneic CD8+ and CD4+ T cells at low DC: T cell ratios. However, at high DC: T cell ratios the stimulatory capacity of Ad-transduced DC was suppressed. This immunosuppressive effect was confirmed by demonstrating that the stimulatory function of untreated DC could be suppressed in a dose-dependent manner by addition of Ad-transduced DC. Furthermore, transwell experiments suggested that direct cell contact was required. Taken together, our results demonstrate the feasibility of efficiently expressing antigens in CD83+ DC using adenoviruses. However, immunosuppressive effects must be considered and carefully studied before Ad-transduced DC are employed for clinical trials. Gene Therapy (2000) 7, 249-254.
我们已经开发出一种用于在无外源血清条件下高效生成具有高度免疫刺激作用的CD83+人树突状细胞（DC）的培养方法。在本研究中，我们评估了使用腺病毒载体在成熟DC中内源性表达抗原的可行性及后果。用Ad-EGFP转导DC后，在50%-85%的CD83+ DC中观察到内源性荧光。Ad转导的DC在低DC:T细胞比例下可刺激同种异体CD8+和CD4+ T细胞增殖。然而，在高DC:T细胞比例下，Ad转导的DC的刺激能力受到抑制。通过证明添加Ad转导的DC可剂量依赖性地抑制未处理DC的刺激功能，证实了这种免疫抑制作用。此外，Transwell实验表明需要直接细胞接触。综上所述，我们的结果证明了使用腺病毒在CD83+ DC中有效表达抗原的可行性。然而，在将Ad转导的DC用于临床试验之前，必须考虑并仔细研究其免疫抑制作用。《基因治疗》（2000年）7卷249 - 254页 。
