Hashimoto Y, Uehara Y, Abu Lila A S, Ishida T, Kiwada H
Department of Pharmacokinetics and Biopharmaceutics, Subdivision of Biopharmaceutical Sciences, Institute of Health Biosciences, The University of Tokushima, Tokushima, Japan.
1] Department of Pharmacokinetics and Biopharmaceutics, Subdivision of Biopharmaceutical Sciences, Institute of Health Biosciences, The University of Tokushima, Tokushima, Japan [2] Department of Pharmaceutics and Industrial Pharmacy, Faculty of Pharmacy, Zagazig University, Zagazig, Egypt.
Gene Ther. 2014 Jun;21(6):593-8. doi: 10.1038/gt.2014.32. Epub 2014 Apr 3.
Cationic liposome represents a promising alternative to viral vectors for the delivery of therapeutic genes. For in vivo use, surface modification of the liposome with polyethylene glycol (PEG) is frequently applied to achieve gene-expression in the targeted tissue. However, we have reported that PEG-coated liposomes have induced anti-PEG IgM, which has caused subsequent doses of PEG-coated liposome to be rapidly cleared from blood circulation, and the complexation of pDNA electrostatically associated with liposome surface has enhanced this antibody response. In this study, we investigated how a Toll-like receptor (TLR) might enhance anti-PEG IgM production. PEG-coated pDNA-lipoplex (PDCL) was injected into either wild type, MyD88 (all TLR adaptor protein, independent of TLR3) knock out (KO) or TLR9 KO mice, and the anti-PEG IgM production levels were detected. Attenuated anti-PEG IgM production following the injection of PDCL was observed in both MyD88 and TLR9 KO mice compared to wild type mice, probably due to the abolished induction of cytokines in both MyD88 and TLR9 KO mice. Our results suggest that TLR, exclusively TLR9, signaling plays a potential role in the enhanced anti-PEG IgM production following the injection of PDCL. This result may have important implications for the design and development of an efficient PEG-coated non-viral gene vector.
阳离子脂质体是一种有前景的替代病毒载体用于递送治疗性基因的载体。对于体内应用，用聚乙二醇（PEG）对脂质体进行表面修饰常用于在靶组织中实现基因表达。然而，我们已经报道过，PEG包被的脂质体可诱导产生抗PEG IgM，这导致后续剂量的PEG包被脂质体从血液循环中迅速清除，并且与脂质体表面静电结合的质粒DNA（pDNA）的复合作用增强了这种抗体反应。在本研究中，我们调查了Toll样受体（TLR）如何增强抗PEG IgM的产生。将PEG包被的pDNA-脂质复合物（PDCL）注射到野生型、髓样分化因子88（MyD88，所有TLR衔接蛋白，不依赖于TLR3）基因敲除（KO）或TLR9基因敲除小鼠体内，并检测抗PEG IgM的产生水平。与野生型小鼠相比，在MyD88和TLR9基因敲除小鼠中注射PDCL后观察到抗PEG IgM产生减弱，这可能是由于MyD88和TLR9基因敲除小鼠中细胞因子的诱导被消除。我们的结果表明，TLR信号传导，尤其是TLR9信号传导，在注射PDCL后增强抗PEG IgM产生中发挥潜在作用。这一结果可能对高效PEG包被的非病毒基因载体的设计和开发具有重要意义。
