Cui Zhengrong, Baizer Lawrence, Mumper Russell J
Division of Pharmaceutical Sciences, College of Pharmacy, University of Kentucky, Lexington, KY 40536-0082, USA.
J Biotechnol. 2003 Apr 24;102(2):105-15. doi: 10.1016/s0168-1656(03)00029-4.
A high population of dendritic cells in the skin makes intradermal (ID) immunization an attractive route. We sought to further enhance immune responses from a previously reported novel nanoparticle-based DNA vaccine delivery system by administering the system intradermally into mouse skin using Biojector 2000, a needle-free jet injection device. Two mouse studies were carried out. Balb/C mice (n=5-6) were immunized on day 0, 7, and 14 by subcutaneous injection or via the Biojector 2000 with pDNA alone (CMV-beta-galactosidase, 5 micro g), pDNA-coated nanoparticles, or beta-galactosidase protein (10 micro g) adjuvanted with 'Alum' (15 micro g). On day 28, mice were sacrificed and specific serum IgG and IgA titer, in vitro cytokine release, and cell proliferation of isolated splenocytes were determined. Similar to previous reports, in both mouse studies, SC immunization with pDNA-coated nanoparticles led to over a log increase in specific serum IgG titer as compared to immunization with pDNA alone. For pDNA alone, jet and SC injection did not result in significant differences in IgG titer. In contrast, for pDNA-coated nanoparticles, jet injection led to as high as a 20-fold enhancement in IgG titer over SC injection. In addition, jet injection of pDNA-coated nanoparticles enhanced the IgG titer by more than 200-fold over jet injection of pDNA alone. Also, jet injection of pDNA-coated nanoparticles resulted in significantly enhanced specific serum IgA titer. For in vitro cytokine release, immunization with pDNA-coated nanoparticles by jet injection enhanced IFN-gamma and IL-4 release over pDNA alone by 6- and 5-fold, respectively. SC injection of pDNA-coated nanoparticles also resulted in enhanced IFN-gamma and IL-4 release over pDNA alone although with less magnitude. Finally, immunization with pDNA-coated nanoparticles, by both jet injection and SC injection, led to improved splenocyte proliferation over pDNA alone. In conclusion, a combination of a novel cationic nanoparticle-based DNA delivery system with ID jet injection led to enhanced antibody production, Th-1/Th-2 balanced cytokine release, and enhanced splenocyte proliferation.
皮肤中大量的树突状细胞使得皮内（ID）免疫成为一种有吸引力的途径。我们试图通过使用无针喷射注射装置Biojector 2000将先前报道的基于新型纳米颗粒的DNA疫苗递送系统皮内注射到小鼠皮肤中，进一步增强免疫反应。进行了两项小鼠研究。在第0、7和14天，通过皮下注射或使用Biojector 2000，用单独的pDNA（CMV-β-半乳糖苷酶，5μg）、pDNA包被的纳米颗粒或与“明矾”（15μg）佐剂联合的β-半乳糖苷酶蛋白（10μg）对Balb/C小鼠（n = 5 - 6）进行免疫。在第28天，处死小鼠并测定特异性血清IgG和IgA滴度、体外细胞因子释放以及分离的脾细胞的增殖情况。与先前的报道相似，在两项小鼠研究中，与单独用pDNA免疫相比，用pDNA包被的纳米颗粒进行皮下免疫导致特异性血清IgG滴度增加超过一个对数。对于单独的pDNA，喷射注射和皮下注射在IgG滴度上没有显著差异。相比之下，对于pDNA包被的纳米颗粒，喷射注射导致的IgG滴度比皮下注射高出20倍。此外, 与单独喷射注射pDNA相比，喷射注射pDNA包被的纳米颗粒使IgG滴度提高了200多倍。而且，喷射注射pDNA包被的纳米颗粒导致特异性血清IgA滴度显著提高。对于体外细胞因子释放，通过喷射注射用pDNA包被的纳米颗粒免疫比单独用pDNA分别使IFN-γ和IL-4释放增加了6倍和5倍。皮下注射pDNA包被的纳米颗粒也导致IFN-γ和IL-4释放比单独用pDNA增加，尽管幅度较小。最后, 通过喷射注射和皮下注射用pDNA包被的纳米颗粒进行免疫均导致脾细胞增殖比单独用pDNA有所改善。总之，一种新型的基于阳离子纳米颗粒的DNA递送系统与皮内喷射注射相结合，导致抗体产生增强、Th-1/Th-2平衡的细胞因子释放以及脾细胞增殖增强。
