(iCeMS), Kyoto University Title - Institute for Integrated Cell

iCeMS | Paper Abstract
Title:
Immunostimulatory characteristics induced by linear polyethyleneimine/plasmid
DNA complexes in cultured macrophages
Author:
Yasunori Saito, Yuriko Higuchi, Shigeru Kawakami, Fumiyoshi Yamashita, Mitsuru
Hashida
Journal:
Human Gene Therapy, Vol. 20, No. 2, 137-145 (2008)
Abstract: Intravenously injected plasmid DNA (pDNA) complexed with cationic liposome
(lipoplexes) caused NFkappaB mediated cytokine production from macrophages
induced by CpG sequence in pDNA. Recently, we have reported that the cytokine
production caused by linear PEI/pDNA complexes (PEI polyplexes) was much
lower than lipoplexes (Kawakami et al., 2006). As Toll like receptor 9 recognizing
CpG sequence is expressed on the endosomal compartment, we hypothesized
that the buffering capacity of PEI enhanced escaping from endosome,
consequently cytokine production was decreased. In this study, comparing with
lipoplexes, mechanism of lower cytokines production induced by PEI polyplexes
were investigated using murine macrophage like cell line RAW 264.7 cells.
Although transfection efficacy and cellular association were similar in PEI
polyplexes and lipoplexes, TNF-alpha and IL-6 production and NFkappaB
activation caused by polyplexes were significantly lower than lipoplexes. As for
intracellular distribution, PEI polyplexes spread in cytosol although lipoplex was
accumulated in vesicles, suggesting enhancement of escape from endosome by
PEI. Bafilomycin A1, inhibitor for of early endosome, enhanced cytokine
production and NFkappaB activation by PEI polyplex but not by lipoplexes,
however chloroquine, inhibitor of late endosome, inhibited PEI polyplexes induced
cytokines production and NFkappaB activation, suggesting that buffering effect of
PEI on early endosome decreases NFkappaB mediated cytokines production. In
conclusion, we demonstrate that the cytokine production and NFkappaB
activation induced by PEI polyplexes are significantly lower than these of
lipoplexes in cultured macrophages. The significant low cytokine response of PEI
polyplexes may be due to effective transition of PEI polyplexes from endosomes
to cytosol.
＜参考訳＞
Title:
培養マクロファージにおける直鎖型ポリエチレンイミン/プラスミド DNA 複合体による免
疫刺激特性
Author:
斎藤泰紀、樋口ゆり子、川上茂、山下富義、橋田充
Journal:
Human Gene Therapy, Vol. 20, No. 2, 137-145 (2008)
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Institute for Integrated Cell-Material Sciences (iCeMS), Kyoto University
iCeMS | Paper Abstract
Abstract: 以前我々は、マウスを用いて直鎖型ポリエチレンイミン/プラスミド DNA 複合体の自然
免疫誘導効果がカチオン性リポソーム/プラスミド DNA 複合体と比べて有意に低いこと
を報告した。そこで、培養マクロファージにおける直鎖型ポリエチレンイミン/プラスミド
DNA 複合体による免疫刺激特性を詳細に解析した。その結果、初期エンドソームにお
ける直鎖型ポリエチレンイミンが有する pH 緩衝作用が直鎖型ポリエチレンイミン/プラ
スミド DNA 複合体のサイトカイン産生作用を阻害しており、その結果、サイトカイン産
生が低くなっている可能性を示すことができた。
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Institute for Integrated Cell-Material Sciences (iCeMS), Kyoto University