Department of Infectious Diseases, College of Veterinary Medicine, Seoul, Republic of Korea.
Eur J Pharm Biopharm. 2011 Oct;79(2):226-31. doi: 10.1016/j.ejpb.2011.04.016. Epub 2011 May 6.
Diarrhea in newborn and weaned piglets is mainly induced by enterotoxigenic Escherichia coli (ETEC) with fimbriae F4 (K88) and F18 (F107). In this study, we evaluated F4 and F18 coated with thiolated Eudragit microspheres (TEMS) as a candidate for an oral vaccine. The average particle sizes of TEMS, F4-loaded TEMS, and F18-loaded TEMS were measured as 4.2±0.75 μm, 4.7±0.50 μm, and 4.5±0.37 μm, respectively. F4 is more efficiently encapsulated than F18 in the loading with TEMS. In the release test, F4 and F18 fimbriae were protected in acidic circumstances, whereas most were released at pH 7.4 of intestine circumstances. Production of TNF-α and NO from RAW 264.7 cells was increased in a time-dependent manner after exposure to all groups, whereas only F4- or F18-loaded TEMS-stimulated IL-6 secretion. The levels of IFN-γ from mouse splenocytes after exposure to F4 or F18 were increased while IL-4 was not detectable. These results suggest that F4- and F18-loaded TEMS may effectively induce immune response with the efficient release of antigens to appropriate target sites.
新生仔猪和断奶仔猪的腹泻主要是由带有菌毛 F4(K88)和 F18(F107)的肠产毒性大肠杆菌(ETEC)引起的。在本研究中,我们评估了用巯基化的 Eudragit 微球(TEMS)包被的 F4 和 F18 作为口服疫苗的候选物。TEMS、F4 负载的 TEMS 和 F18 负载的 TEMS 的平均粒径分别测量为 4.2±0.75μm、4.7±0.50μm 和 4.5±0.37μm。与 TEMS 负载相比,F4 更有效地被包封。在释放试验中,F4 和 F18 菌毛在酸性环境中得到保护,而在肠道环境的 pH7.4 时大部分被释放。暴露于所有组后,RAW264.7 细胞中 TNF-α和 NO 的产生呈时间依赖性增加,而只有 F4 或 F18 负载的 TEMS 刺激 IL-6 的分泌。暴露于 F4 或 F18 后,小鼠脾细胞中 IFN-γ的水平增加,而 IL-4 不可检测。这些结果表明,F4 和 F18 负载的 TEMS 可以有效地诱导免疫反应,并将抗原有效地释放到适当的靶位。