University Medical Center of the Johannes Gutenberg-University, Clinical Research Unit Allergology, Department of Dermatology, Mainz, Germany.
J Gene Med. 2010 Mar;12(3):231-43. doi: 10.1002/jgm.1436.
Dendritic cells (DCs) constitute an attractive target for immunotherapeutic approaches. Because DCs are largely refractory to transfection with plasmid DNA, several viral transduction protocols were established. The potential side-effects of lentiviral transduction on the phenotype and activation state of DCs left unstimulated after transduction have not been assessed. There is a need to analyse these parameters as a result of the requirement of using DCs with a low activation state for therapeutic strategies intended to induce tolerance.
Lentivirally-transduced bone marrow (BM)-derived DCs (LV-DCs) in comparison with mock-transduced (Mock-DCs) and untreated DCs were analysed with regard to the induction of maturation processes on the RNA, protein and functional level. BM-DCs engineered to overexpress interleukin (IL)-10 were analysed for therapeutic potential in a mouse model of allergic contact dermatitis.
Compared with untreated DCs, Mock-DCs and LV-DCs displayed an altered gene expression signature. Mock-DCs induced a stronger T cell proliferative response than untreated DCs. LV-DCs did not further augment the T cell proliferative response, but induced a slightly different T cell cytokine pattern compared to Mock-DCs. Accordingly, the gene promoter of the DC maturation marker fascin mediated efficient expression of the model transgene IL-10 in unstimulated-transduced BM-DCs. Nevertheless, IL-10 overexpressing BM-DCs exerted tolerogenic activity and efficiently inhibited the contact hypersensitivity response in previously hapten-sensitized mice.
Lentiviral transduction of BM-DCs results in their partial activation. Nevertheless, the transduction of these DCs with a vector encoding the immunomodulatory cytokine IL-10 rendered them tolerogenic. Thus, lentivirally-transduced DCs expressing immunomodulatory molecules represent a promising tool for induction of tolerance.
树突状细胞（DCs）是免疫治疗方法的一个有吸引力的靶标。由于 DCs 对质粒 DNA 的转染基本没有反应，因此建立了几种病毒转导方案。未经刺激的转导后，慢病毒转导对 DCs 表型和激活状态的潜在副作用尚未得到评估。由于需要使用处于低激活状态的 DC 来进行旨在诱导耐受的治疗策略，因此需要分析这些参数。
与模拟转染（Mock-DCs）和未处理的 DC 相比，分析了慢病毒转染的骨髓（BM）来源的 DC（LV-DCs）在 RNA、蛋白质和功能水平上诱导成熟过程的情况。分析了过表达白细胞介素（IL）-10 的 BM-DC 用于治疗变应性接触性皮炎的小鼠模型的治疗潜力。
与未处理的 DC 相比，Mock-DCs 和 LV-DCs 显示出改变的基因表达特征。Mock-DCs 诱导更强的 T 细胞增殖反应，而未处理的 DC 则较弱。LV-DCs 没有进一步增强 T 细胞的增殖反应，但与 Mock-DCs 相比，诱导了略有不同的 T 细胞细胞因子模式。相应地，DC 成熟标志物 fascin 的基因启动子在未刺激转导的 BM-DC 中有效地表达了模型转基因 IL-10。然而，过表达 IL-10 的 BM-DC 发挥了耐受原活性，并有效地抑制了先前敏化小鼠的接触超敏反应。
BM-DC 的慢病毒转导导致其部分激活。然而，用编码免疫调节细胞因子 IL-10 的载体转导这些 DC 使其具有耐受原性。因此，表达免疫调节分子的慢病毒转导 DC 代表诱导耐受的有前途的工具。
