Enhanced in vivo gene expression mediated by listeriolysin O incorporated anionic LPDII: Its utility in cytotoxic T lymphocyte-inducing DNA vaccine.
Enhanced in vivo gene expression mediated by listeriolysin O incorporated anionic LPDII: Its utility in cytotoxic T lymphocyte-inducing DNA vaccine.
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DOI：
10.1016/j.jconrel.2010.06.017
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发表时间：
2010-12-01
期刊：
影响因子：
--
通讯作者：
Lee KD
中科院分区：
文献类型：
--
作者：
Sun X;Provoda C;Lee KD
一般来说，利用非病毒载体增强体内基因表达是基因治疗中的一个关键问题。在非病毒载体介导的基因传递的许多潜在用途中，它在基于dna的疫苗接种中的应用是一种有吸引力的方法，比传统疫苗接种具有几个实际优势。我们之前已经证明，内溶细菌蛋白李斯特菌溶素O （LLO）能够通过LPDII（阴离子脂质体-多阳离子- dna复合物）传递系统促进体外转染。在本研究中，我们扩展并研究了含有低氧化酶的LPDII在体内的DNA递送，并评估了其在小鼠DNA疫苗接种中的效用。我们进一步研究了这种非病毒基因传递系统在传递含有OVA cDNA的质粒后引发对模型抗原卵清蛋白（OVA）的免疫反应的能力，特别是关注OVA特异性CD8+细胞毒性T淋巴细胞（CTL）反应。采用DNA引物和蛋白质增强方案来产生细胞毒性T细胞反应。我们的研究结果表明，当LLO加入到LPDII中时，观察到体外和体内转染效率的提高。与热灭活的含有低密度脂蛋白的LPDII （hi -低密度脂蛋白）制剂相比，这种低密度脂蛋白制剂在体内产生了增强的功能性抗原特异性CD8+ T细胞反应。此外，通过酶联免疫吸附斑点试验，在从小鼠分离的脾细胞中观察到明显更高的CTL频率。在ova特异性CD8+肽的特异性刺激下，当LLO包含在LPDII中时，干扰素γ的产生也显著增强。这些发现表明，含有低分子量脂蛋白的LPDII系统作为DNA疫苗递送的候选载体具有显著的潜力。
Enhanced in vivo gene expression using non-viral vectors is a critical issue in gene therapy in general. Among the many potential utilities of non-viral vector-mediated gene delivery, its application in DNA-based vaccination is an attractive approach with several practical advantages over conventional vaccination. We have previously shown that the endosomolytic bacterial protein listeriolysin O (LLO) is capable of facilitating transfection in vitro using the LPDII (anionic liposome-polycation-DNA complexes) delivery system. In the present study we have extended and investigated the DNA delivery of LLO-containing LPDII to in vivo and evaluated its utility in DNA vaccination in mice. We further investigated the ability of this non-viral gene delivery system to elicit an immune response to a model antigen ovalbumin (OVA), particularly focusing on the OVA-specific CD8+ cytotoxic T lymphocyte (CTL) response, after delivery of a plasmid containing the OVA cDNA. A DNA prime and protein boost protocol was employed to generate cytotoxic T cell responses. Our results show that increased in vitro and in vivo transfection efficiencies were observed when LLO was incorporated into LPDII. This LLO-LPDII formulation produced an enhanced functional antigen-specific CD8+ T cell response in vivo compared to the heat-inactivated LLO-containing LPDII (HI-LLO-LPDII) formulation. Furthermore, a significantly higher CTL frequency was observed in the splenocytes isolated from the mice primed with LLO-LPDII by an enzyme-linked immunosorbent spot assay. Interferon-γ production upon specific stimulation by OVA-specific CD8+ peptide was also significantly stronger with the inclusion of LLO into LPDII. These findings suggest that the LLO-containing LPDII system possesses noteworthy potential as a candidate carrier for DNA vaccine delivery.
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DOI：
10.1038/nrg2432
发表时间：
2008-10
期刊：
Nature reviews. Genetics
影响因子：
--
作者：
通讯作者：
--
影响因子：
5.1
作者：
Eliyahu, H;Servel, N;Barenholz, Y
通讯作者：
Barenholz, Y
影响因子：
5.8
作者：
Huygen, K
通讯作者：
Huygen, K
影响因子：
4.8
作者：
Lee, KD;Oh, YK;Swanson, JA
通讯作者：
Swanson, JA
DOI：
10.1084/jem.186.7.1159
发表时间：
1997-10-06
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Beauregard KE;Lee KD;Collier RJ;Swanson JA
通讯作者：
Swanson JA