Upconversion nanoparticles modified with aminosilanes as carriers of DNA vaccine for foot-and-mouth disease
Upconversion nanoparticles modified with aminosilanes as carriers of DNA vaccine for foot-and-mouth disease
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氨基硅烷修饰的上转换纳米颗粒作为口蹄疫 DNA 疫苗的载体
DOI：
10.1007/s00253-012-4042-z
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发表时间：
2012-09-01
影响因子：
5
通讯作者：
Liu, Xiangtao
中科院分区：
文献类型：
--
作者：
Guo, Huichen;Hao, Rongzeng;Liu, Xiangtao
以口蹄疫病毒(FMDVAsiaI/江苏2005)DNA疫苗为载体，评价了上转换纳米NaYF4：Yb/Er@SiO_2(UCPs)/质粒DNA(pcDNA3.1/VP1-GFP)复合体诱导免疫应答的潜力。采用体外抑制法检测UCPs对DNaseI降解的保护作用。用激光共聚焦显微镜检测其在体内的表达。通过细胞毒性实验评价其生物相容性。基于这一结果，氨基硅烷修饰的UCPs能够静电结合、凝聚和保护质粒DNA。细胞存活率分析表明，UCPs/质粒DNA复合体的细胞毒性低于阳离子脂/质粒DNA复合体，而UCPs的转染率与阳离子脂类相当。此外，将UCP/质粒DNA复合体肌肉注射给豚鼠。用间接酶联免疫吸附试验(ELISA)、微量中和试验和T淋巴细胞增殖试验检测体液和细胞免疫应答。免疫后可诱导抗口蹄疫病毒特异性抗体、中和抗体和T淋巴细胞增殖反应。在攻击试验中，所有接种UCPs/质粒DNA复合体的豚鼠都能完全抵抗口蹄疫病毒的攻击。目前的研究鼓励使用UCPs作为一种有效的纳米系统，用于将基因输送到细胞，用于体外和体内疫苗接种，以及其他治疗应用。
The potential of the upconversion nanoparticles NaYF4:Yb/Er@silica(UCPs)/plasmid DNA (pcDNA3.1/VP1-GFP) complex in inducing immune responses was evaluated using the UCPs as carriers of the foot-and-mouth disease virus (FMDVAsiaI/Jiangsu2005) DNA vaccine. The UCPs protection against DNaseI degradation was measured using an in vitro inhibition assay. The expression of the plasmid in vivo was determined via confocal microscopy. Its biocompatibility was evaluated through cytotoxicity assay. Based on the results, the aminosilane-modified UCPs can electrostatically bind, condense, and protect plasmid DNA. Cell viability assays demonstrated that the cytotoxicity of the UCPs/plasmid DNA complex is lower than that of the cationic lipid/plasmid DNA complex, and that the transfection efficiency of UCPs is the same as that of the cationic lipid. Furthermore, the UCP/plasmid DNA complex was intramuscularly administered to guinea pigs. Humoral and cellular immune responses were detected using indirect enzyme-linked immunosorbent assay (ELISA), micro-neutralization assay, and T-lymphocyte proliferation assay. Anti-FMDV specific antibodies, neutralizing antibodies, and T-lymphocyte proliferation responses were induced after vaccination. In the challenge test, all the guinea pigs vaccinated with the UCPs/plasmid DNA complex were fully protected from the FMDV challenge. The current study encourages the use of UCPs as an effective nanosystem for gene delivery to cells for in vitro and in vivo vaccination, and other therapeutic applications.