Activation of immature monocyte-derived dendritic cells after transduction with high doses of lentiviral vectors
Activation of immature monocyte-derived dendritic cells after transduction with high doses of lentiviral vectors
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DOI：
10.1089/hum.2007.006
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发表时间：
2007-06-01
影响因子：
4.2
通讯作者：
Thielemansi, Kris
中科院分区：
文献类型：
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作者：
Breckpot, Karine;Emeagi, Perpetua;Thielemansi, Kris
树突状细胞(DC)是一种很有吸引力的免疫调节工具，它针对成熟的DC(MDCs)进行免疫或针对未成熟/半成熟的DC(IDCs)进行耐受。因此，将抗原引入树突状细胞已成为各种免疫学学科的首要课题。大量研究表明，慢病毒是实现这一目的的有效载体。本研究评估慢病毒转导对IDC激活的影响。随着慢病毒剂量的增加，未成熟的DC可以有效地被转导，而不会影响细胞的存活。低感染复数(MOI)的转导不会导致表型或功能成熟。然而，更高剂量的慢病毒导致黏附分子、共刺激分子和人类白细胞抗原分子的上调，以及同种异体刺激能力和IL-6、IL-8和肿瘤坏死因子-a的分泌增加。只有在极高剂量下才能观察到IL-12p70、IL-10和干扰素-a的产生。Western blotting显示，在MOI为150的情况下，蛋白激酶R被磷酸化。Toll样受体(TLR)驱动的荧光素酶报告实验显示，TLR2、TLR3和TLR8的激活具有剂量依赖性，与伪型、生产或转导方案无关，并在热失活时被取消。这些数据表明，慢病毒载体不仅为IDCs提供抗原，而且还提供适当的激活信号，有利于将其用于免疫治疗和疫苗开发。
Dendritic cells (DCs) are an attractive tool for immunomodulation, targeting mature DCs (mDCs) for immunization or immature/semimature DCs (iDCs) for tolerization. Therefore, introducing antigens into DCs has become a prime topic in various immunological disciplines. Numerous studies have shown that lentiviruses are an efficient vehicle for this purpose. This study evaluates the effects of lentiviral transduction on iDC activation. Immature DCs are efficiently transduced with increasing doses of lentivirus without affecting cell viability. Transduction at low multiplicities of infection (MOIs) did not result in phenotypical or functional maturation. Higher doses of lentivirus, however, resulted in upregulation of adhesion, costimulatory, and HLA molecules, as well as in increased allostimulatory capacity and secretion of interleukin (IL)-6, IL-8, and tumor necrosis factor-a. Production of IL-12 p70, IL-10, and interferon-a was observed only at extremely high doses. Protein kinase R phosphorylation on transduction at an MOI of 150 was demonstrated by Western blotting. A Toll-like receptor (TLR)-driven luciferase reporter assay showed dose-dependent activation of TLR2, TLR3, and TLR8, which was independent of the pseudotype, production, or transduction protocol and was abrogated on heat inactivation. These data show that lentiviral vectors provide not only the antigen but also appropriate activation signals to iDCs, favoring their use for immunotherapy and vaccine development.