Center for NanoBiotechnology Research, Alabama State University, Montgomery, Alabama, USA.
Nanomedicine. 2009 Dec;5(4):463-72. doi: 10.1016/j.nano.2009.02.004. Epub 2009 Mar 31.
This study evaluated the efficiency of chitosan-encapsulated DNA-based respiratory syncytial virus (RSV) vaccine. Antigenic regions of RSV F, M2, and G genes were cloned into the human cytomegalovirus promoter-based constitutive expression vector, resulting in a DNA vaccine vector named DR-FM2G. This vector was used to formulate DNA-chitosan nanoparticles (DCNPs) using a complex coacervation process that yielded an encapsulation efficiency of 94.7%. The DCNP sizes ranged from 80 to 150 nm with uniform size distribution and spherical shape. DNA release was between 50% and 60% when DCNPs were incubated with similar gastrointestinal fluid (pH 2), whereas 21% to 25% of DNA was released from DCNPs in 30 minutes at pH 10. Differential scanning calorimetry showed DCNPs to be more stable than naked DNA or chitosan, offering protection from DNA degradation by nucleases. DCNPs were not toxic to cells when used at concentrations < or =400 microg/mL. Immunohistochemical and real-time polymerase chain reaction results showed a higher level of RSV protein expression in mouse tissues given when DCNPs were injected intravenously as compared with naked DNA.
This study evaluated the efficiency of chitosan-encapsulated DNA-based respiratory syncytial virus (RSV) vaccine, showing a higher level of RSV protein expression in mouse tissues given when DCNPs were injected intravenously as compared with naked DNA.
本研究评估了壳聚糖包封的 DNA 基于呼吸道合胞病毒（RSV）疫苗的效率。RSV F、M2 和 G 基因的抗原区域被克隆到人巨细胞病毒启动子组成型表达载体中，得到一种名为 DR-FM2G 的 DNA 疫苗载体。该载体用于通过复杂共凝聚过程来制备 DNA-壳聚糖纳米颗粒（DCNP），得到的包封效率为 94.7%。DCNP 的粒径范围为 80 至 150nm，具有均匀的粒径分布和球形。当 DCNP 在类似的胃肠液（pH2）中孵育时，DNA 释放量在 50%至 60%之间，而在 pH10 时，21%至 25%的 DNA 从 DCNP 中释放出来。差示扫描量热法表明 DCNP 比裸露 DNA 或壳聚糖更稳定，能够保护 DNA 免受核酸酶的降解。当使用浓度<或=400μg/ml 时，DCNP 对细胞没有毒性。免疫组织化学和实时聚合酶链反应结果表明，与裸露 DNA 相比，静脉内注射 DCNP 时，小鼠组织中 RSV 蛋白的表达水平更高。
本研究评估了壳聚糖包封的 DNA 基于呼吸道合胞病毒（RSV）疫苗的效率，与裸露 DNA 相比，静脉内注射 DCNP 时，小鼠组织中 RSV 蛋白的表达水平更高。
