Bos Gert W, Kanellos Theofanis, Crommelin Daan J A, Hennink Wim E, Howard Colin R
Department of Pharmaceutics, Faculty of Pharmacy, Utrecht Institute for Pharmaceutical Sciences (UIPS), Utrecht University, P.O. Box 80.082, 3508 TB Utrecht, The Netherlands.
Vaccine. 2004 Dec 9;23(4):460-9. doi: 10.1016/j.vaccine.2004.06.020.
In this paper, different cationic polymers were investigated as a DNA delivery system both in vitro in dendritic and muscle cells and in vivo, in a murine model. Expression of the reporter gene beta-galactosidase was used in order to determine the in vitro transfection efficiency of these polymer-DNA complexes (polyplexes) and both specific mRNA and protein expression were monitored in parallel with polyplex toxicity on the cells. Interestingly, the enhancing expression activities of the different polyplexes were tissue-dependent, implying that they may gain entrance to the cells through specific receptors. Subsequently, complexes of polymers and DNA plasmid (pCMV-S) encoding the human hepatitis B virus (HBV) surface antigen (HBsAg) were injected into the skeletal muscles of BALB/c mice. Higher levels of both HBsAg local expression in the tibial anterior muscles and systemic humoral immune responses were detected when the selected polymers complexed with pCMV-S were compared to those complexed with pCMV-S alone. Induction of immunoglobulin G2a (IgG2a) against HbsAg in the serum of pCMV-S-polyplex vaccinated mice varied with the polymer used, suggesting that polyplex-mediated DNA vaccination can potentially modulate the type of helper T cell immunity (Th). The effect of some polyplexes to switch the host immune response more towards a Th1 response may be associated with their differential efficiency to transfect dendritic cells and/or other antigen-presenting cells (APC) as was observed in vitro. These results suggest that the investigated cationic polymers can be effective as delivery/adjuvant compounds for DNA.
在本文中，研究了不同的阳离子聚合物作为DNA递送系统在树突状细胞和肌肉细胞中的体外情况以及在小鼠模型中的体内情况。使用报告基因β-半乳糖苷酶的表达来确定这些聚合物-DNA复合物（多聚体）的体外转染效率，并同时监测特定mRNA和蛋白质表达以及多聚体对细胞的毒性。有趣的是，不同多聚体的增强表达活性具有组织依赖性，这意味着它们可能通过特定受体进入细胞。随后，将编码人乙型肝炎病毒（HBV）表面抗原（HBsAg）的聚合物与DNA质粒（pCMV-S）的复合物注射到BALB/c小鼠的骨骼肌中。当将与pCMV-S复合的选定聚合物与单独与pCMV-S复合的聚合物进行比较时，检测到胫骨前肌中HBsAg的局部表达水平和全身体液免疫反应均更高。在接种pCMV-S-多聚体的小鼠血清中，针对HbsAg的免疫球蛋白G2a（IgG2a）的诱导随所用聚合物的不同而变化，这表明多聚体介导的DNA疫苗接种可能潜在地调节辅助性T细胞免疫（Th）的类型。如在体外观察到的那样，一些多聚体将宿主免疫反应更多地转向Th1反应的作用可能与其转染树突状细胞和/或其他抗原呈递细胞（APC）的不同效率有关。这些结果表明，所研究的阳离子聚合物可作为DNA的有效递送/佐剂化合物。
