Basner-Tschakarjan Etiena, Gaffal Evelyn, O'Keeffe Meredith, Tormo Damia, Limmer Andreas, Wagner Hermann, Hochrein Hubertus, Tüting Thomas
Laboratory of Experimental Dermatology, Department of Dermatology, University of Bonn, Germany.
J Gene Med. 2006 Nov;8(11):1300-6. doi: 10.1002/jgm.964.
Recombinant replication-deficient adenoviral vectors (recAd) are attractive candidates for DNA vaccination approaches because they are able to activate the innate and adaptive immune systems. Here we explore the ability of recAd to transduce and activate subsets of dendritic cells, namely plasmacytoid dendritic cells (pDC) and conventional dendritic cells (cDC).
DC were derived from bone marrow precursors in vitro with the help of FLT3-ligand. Sorted populations of pDC and cDC were infected with recAd at various multiplicities of infection. Transduction efficiency, phenotypic maturation and production of IFN-alpha as well as IL-6 were assessed. Additionally, activation of DC and induction of cytotoxic T lymphocytes (CTL) were determined in vivo. The role of Toll-like receptor (TLR) 9 in recAd recognition was investigated as it has previously been shown that DNA viruses are recognized via this receptor.
RecAd can efficiently transduce pDC as well as cDC in vitro. Both DC subsets mature and produce IFN-alpha upon interaction with recAd. In the absence of TLR9, activation and cytokine production was only detected in cDC but not in pDC. Importantly, induction of CD8+ CTL following in vivo injection of recAd was similar in TRL9-deficient mice when compared with wildtype controls.
RecAd can efficiently transduce and activate both pDC and cDC. pDC required TLR9 to detect the presence of recAd whereas cDC also recognized recAd independently of TLR9. These unique immunostimulatory properties support the future development of recombinant Ad as a vector for DNA vaccine approaches.
重组复制缺陷腺病毒载体（recAd）是DNA疫苗接种方法中颇具吸引力的候选者，因为它们能够激活先天性和适应性免疫系统。在此，我们探究recAd转导并激活树突状细胞亚群，即浆细胞样树突状细胞（pDC）和传统树突状细胞（cDC）的能力。
在FLT3配体的帮助下，体外从骨髓前体细胞中获得树突状细胞。以不同的感染复数用recAd感染分选后的pDC和cDC群体。评估转导效率、表型成熟以及IFN-α和IL-6的产生。此外，在体内测定树突状细胞的激活以及细胞毒性T淋巴细胞（CTL）的诱导。研究了Toll样受体（TLR）9在recAd识别中的作用，因为先前已表明DNA病毒通过该受体被识别。
RecAd在体外能有效转导pDC以及cDC。两个树突状细胞亚群在与recAd相互作用后均成熟并产生IFN-α。在缺乏TLR9的情况下，仅在cDC中检测到激活和细胞因子产生，而在pDC中未检测到。重要的是，与野生型对照相比，在体内注射recAd后，TLR9缺陷小鼠中CD8 + CTL的诱导相似。
RecAd能有效转导并激活pDC和cDC。pDC需要TLR9来检测recAd的存在，而cDC也能独立于TLR9识别recAd。这些独特的免疫刺激特性支持了重组腺病毒作为DNA疫苗接种方法载体的未来发展。
