Lipoplex-induced hemagglutination: potential involvement in intravenous gene delivery
Lipoplex-induced hemagglutination: potential involvement in intravenous gene delivery
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DOI：
10.1038/sj.gt.3301705
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发表时间：
2002-07-01
期刊：
影响因子：
5.1
通讯作者：
Barenholz, Y
中科院分区：
文献类型：
--
作者：
Eliyahu, H;Servel, N;Barenholz, Y
我们报告了一项研究，旨在表征血液和血液成分与脂质体在体内静脉转染相关条件下的相互作用。在这项研究中，我们专注于脂质复合物与红细胞（RBC）的相互作用。发现在使用通常用于体内转染的范围内的lipoplex组合物和lipoplex/红细胞比率的几小时温育期间没有发生显著的溶血。然而，红细胞与脂质复合物的相互作用导致大量凝集，这与阳离子脂质或辅助脂质的类型无关。在亚精胺或硫酸鱼精蛋白存在下，聚合复合物（如树枝状聚合物/DNA复合物）和脂质复合物也诱导凝集（后者本身诱导血凝）。插入脂质复合物中的DSPE-PEG（2000）在一定程度上抑制血凝。为了更好地了解血清对凝集的影响，将血浆分离成其高分子量组分（HMWC，> 14 kDa）和其低分子量组分（LMWC，小于或等于514 kDa）。这些馏分的特点是他们的蛋白质水平，伯氨基，渗透压和电导率，并与盐水（0.15 M NaCl）进行比较。结果发现，LMWC和HMWC本身都抑制凝集，尽管全血清表现出比每个组分单独更好的血凝抑制作用。血清（或血浆）的抑制作用是通过其对脂质复合物的静电的影响来解释的，减少了它们的正电荷，如使用荧光素-磷脂酰乙醇胺标记的脂质复合物所证明的。LMWC的效果与离子强度有关，相当于0.15 M NaCl的效果。凝集水平随着lipoplex DNA（-）/阳离子脂质（DNA（-）/L+）比率的增加而降低。然而，在静脉内给药后实现显著体内转染所需的低DNA（-）/L+比率下，发生大量凝集。这些数据表明，静脉注射脂质复合物和复合物可能会导致红细胞凝集，形成的凝集物可以解释脂质复合物的定位及其转基因在肺中的表达。
We report a study aiming to characterize the interaction of blood and blood components with lipoplexes under conditions relevant to in vivo intravenous transfection. In this study we focus on the interaction of lipoplexes with red blood cells (RBC). It was found that no significant hemolysis occurred during several hours' incubation using lipoplex compositions and lipoplex/red blood cell ratios in the range commonly used for in vivo transfection. However, the interaction of RBC with lipoplexes resulted in massive agglutination, which occurs irrespective of the type of cationic lipid or helper lipid. Agglutination was also induced by polyplexes (such as dendrimer/DNA complexes) and lipoplexes in the presence of spermidine or protamine sulfate (the latter induced hemagglutination by itself). DSPE-PEG(2000) inserted into the lipoplexes inhibits hemagglutination somewhat. In order to understand the effect of serum on the agglutination better, plasma was separated into its high molecular weight components (HMWC, > 14 kDa) and its low molecular weight components (LMWC, less than or equal to 514 kDa). These fractions were characterized for their level of proteins, primary amino groups, osmotic pressure, and electrical conductivity, and compared with saline (0.15 M NaCl). It was found that both LMWC and HMWC inhibit agglutination by themselves, although whole serum demonstrates better hemagglutination inhibition than each fraction separately. The inhibitory effect of the serum (or plasma) is explained by its effect on the electrostatics of the lipoplexes, reducing their positive charge, as was demonstrated using fluorescein-phosphatidylethanolamine-labeled lipoplexes. The effect of LMWC was related to ionic strength and was equal to the effect of 0.15 M NaCl. The level of agglutination was reduced with increasing lipoplex DNA(-)/cationic lipid (DNA(-)/L+) ratio. However, at the low DNA(-)/L+ ratio needed to achieve significant in vivo transfection after i.v. administration, massive agglutination occurred. These data suggest that i.v. administration of lipoplexes and polyplexes may lead to RBC agglutination, and the agglutinates formed may explain the localization of lipoplexes and expression of their transgenes in the lungs.