Impact of the Charge Ratio on the In Vivo Immunogenicity of Lipoplexes.
Impact of the Charge Ratio on the In Vivo Immunogenicity of Lipoplexes.
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DOI：
10.1007/s11095-017-2187-2
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发表时间：
2017-09
影响因子：
3.7
通讯作者：
Bansal, Geetha P.
中科院分区：
文献类型：
--
作者：
Heidari, Zahra;Arora, Jaspreet S.;Datta, Dibyadyuti;John, Vijay T.;Kumar, Nirbhay;Bansal, Geetha P.
本研究利用编码Pfs25的DNA质粒研究了不同阳离子脂质体制剂的免疫原性潜力，Pfs25是一种疟疾传播阻断候选疫苗。将Pfs25质粒DNA与阳离子脂质体络合，形成阳离子脂质头基团与核苷酸磷酸（N:P）不同电荷比的脂质体。低温透射电镜观察脂质体的形成。用共聚焦显微镜观察GFP编码质粒DNA形成的脂丛，并用流式细胞术检测其体外转染能力。利用编码Pfs25的质粒DNA对Balb/c小鼠肌肉注射后，对两种不同的脂质体制剂进行了体内免疫原性评价。ELISA法检测免疫血清。结果表明，阳离子脂质体介导的N:P电荷比为1:3的DNA免疫（阴离子脂质体）比单独使用裸质粒DNA更有效。当使用N:P电荷比为10:3的阳离子脂质体时，未观察到抗体应答。在一些脂质体制剂中加入海藻糖作为冷冻保护剂和佐剂，但在体内并没有产生任何进一步的免疫原性改善。结果表明，Pfs25质粒DNA以1:3的电荷比作为脂质体在小鼠体内引起了较强的免疫原性，并且可以进一步改善以匹配体内电穿孔给药的DNA疫苗的免疫应答。
The present study investigated the immunogenic potential of different cationic liposome formulations with a DNA plasmid encoding Pfs25, a malaria transmission-blocking vaccine candidate. Pfs25 plasmid DNA was complexed with cationic liposomes to produce lipoplexes at different charge ratios of the cationic lipid head group to the nucleotide phosphate (N:P). The formation of lipoplexes was visualized by Cryogenic-TEM. Confocal microscopy of lipoplexes formed with GFP encoding plasmid DNA, and flow cytometry was used to determine their in vitro transfection capability. Two different lipoplex formulations using plasmid DNA encoding Pfs25 were evaluated for in vivo immunogenicity after intramuscular administration in Balb/c mice. Immune sera were analyzed by ELISA. The results demonstrated that the cationic liposome-mediated DNA immunization with an N:P charge ratio of 1:3 (anionic lipoplexes) is more effective than the use of naked plasmid DNA alone. No antibody response was observed when lipoplexes with a higher N:P charge ratio of 10:3 (cationic lipoplexes) were used. Trehalose was added to some lipoplex formulations as a cryoprotectant and adjuvant, but it did not yield any further improvement of immunogenicity in vivo. The results suggest that Pfs25 plasmid DNA delivered as lipoplexes at a charge ratio of 1:3 elicited strong immunogenicity in mice and may be improved further to match the immune responses of DNA vaccines administered by in vivo electroporation.
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DNA疫苗的电穿孔输送：成功的前景。
影响因子：
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作者：
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通讯作者：
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