Enhanced CTL response by controlled intracellular trafficking of antigen in dendritic cells following DNA vaccination
Enhanced CTL response by controlled intracellular trafficking of antigen in dendritic cells following DNA vaccination
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DNA 疫苗接种后，通过控制树突状细胞中抗原的细胞内运输来增强 CTL 反应
DOI：
10.1016/j.jconrel.2009.01.026
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发表时间：
2009
影响因子：
10.8
通讯作者：
Takakura Y.
中科院分区：
文献类型：
--
作者：
Isaji K;Kawase A;Matono M;Guan X;Nishikawa M;Takakura Y.
为了在DNA疫苗接种后有效地引发细胞毒性T淋巴细胞（CTL）应答，我们构建了几种质粒DNA（pDNA）载体，其编码卵清蛋白（OVA）或具有修饰的细胞内运输的OVA蛋白的主要组织相容性复合物（MHC）I类限制性表位肽（SIINFEKL）。使用DC 2.4细胞（树突状细胞系）进行体外抗原呈递测定，以检查直接转染后构建体的潜力。在这些载体中，pPep-ER（编码抗原肽的pDNA与内质网（ER）滞留信号结合）与没有信号的对应物相比表现出显著的抗原呈递能力。基于体外结果，我们使用pPep-ER通过皮内或肌内途径结合电穿孔在小鼠中进行体内免疫实验。pPep-ER显示出有效的抗原特异性CTL诱导，并且效果上级于阳性对照完全弗氏佐剂（CFA）中的OVA所显示的效果。与无pPep-free相比，pPep-ER显着增加了脾细胞释放的干扰素γ（IFN-γ）水平。用pPep-ER免疫E.G7肿瘤也表现出高度的抑制作用。这些结果表明，用表达具有受控细胞内运输的MHC I类表位肽的pDNA载体进行DNA疫苗接种是通过直接呈递诱导抗原特异性CTL应答的有前景的方法。
To elicit a cytotoxic T lymphocytes (CTL) response efficiently after DNA vaccination, we constructed several plasmid DNA (pDNA) vectors encoding the major histocompatibility complex (MHC) class I-restricted epitope peptide (SIINFEKL) of ovalbumin (OVA) or OVA protein with modified intracellular trafficking. An in vitro antigen presentation assay was carried out using DC2.4 cells, a dendritic cell line, to examine the potentials of the constructs following direct transfection. Among the vectors, pPep-ER, pDNA encoding antigen peptide combined with an endoplasmic reticulum (ER)-retention signal, exhibited a significant ability of antigen presentation compared with the counterpart without the signal. Based on the in vitro results, we carried out in vivo immunization experiments using pPep-ER via the intradermal or intramuscular route in combination with electroporation in mice. pPep-ER showed an efficient antigen-specific CTL induction and the effect was superior to that exhibited by the positive control, OVA in complete Freund's adjuvant (CFA). The levels of interferon gamma (IFN-γ) released from spleen cells were significantly increased by pPep-ER compared with pPep-free. Immunization with pPep-ER also exhibited a high inhibitory effect on the growth of E.G7 tumor. These results indicate that DNA vaccination with the pDNA vector expressing a MHC class I epitope peptide with controlled intracellular trafficking is a promising method of inducing an antigen-specific CTL response via direct presentation.