Molecular Pharmaceutics, Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599-7571, USA.
Pharm Res. 2010 Feb;27(2):350-60. doi: 10.1007/s11095-009-0028-7. Epub 2009 Dec 19.
To study the use of poly (lactide-co-glycolide) (PLGA) microparticles in respirable sizes as carriers for recombinant tuberculosis (TB) antigen, TB10.4-Ag85B, with the ultimate goal of pulmonary delivery as vaccine for the prevention of TB.
Recombinant TB antigens were purified from E. coli by FPLC and encapsulated into PLGA microparticles by emulsion/spray-drying. Spray-drying condition was optimized by half-factorial design. Microparticles encapsulating TB antigens were assessed for their ability to deliver antigens to macrophages for subsequent presentation by employing an in vitro antigen presentation assay specific to an Ag85B epitope.
Spray-drying condition was optimized to prepare PLGA microparticles suitable for pulmonary delivery (aerodynamic diameter of 3.3 microm). Antigen release from particles exhibited an initial burst release followed by sustained release up to 10 days. Antigens encapsulated into PLGA microparticles induced much stronger interleukin-2 secretion in a T-lymphocyte assay compared to antigen solutions for three particle formulations. Macrophages pulsed with PLGA-MDP-TB10.4-Ag85B demonstrated extended epitope presentation.
PLGA microparticles in respirable sizes were effective in delivering recombinant TB10.4-Ag85B in an immunologically relevant manner to macrophages. These results set the foundation for further investigation into the potential use of PLGA particles for pulmonary delivery of vaccines to prevent Mycobacterium tuberculosis infection.
研究聚（乳酸-共-乙醇酸）（PLGA）亚微米颗粒作为重组结核（TB）抗原 TB10.4-Ag85B 的载体的用途，最终目标是通过肺部给药作为预防 TB 的疫苗。
从大肠杆菌中通过 FPLC 纯化重组 TB 抗原，并通过乳液/喷雾干燥将其包封在 PLGA 亚微米颗粒中。通过半因子设计优化喷雾干燥条件。通过体外抗原呈递测定评估包封 TB 抗原的微粒，该测定专门针对 Ag85B 表位，用于评估抗原向巨噬细胞递呈的能力。
优化喷雾干燥条件以制备适合肺部给药的 PLGA 亚微米颗粒（空气动力学直径为 3.3 微米）。颗粒中的抗原释放表现出初始突释，随后持续释放长达 10 天。与三种颗粒制剂的抗原溶液相比，包封在 PLGA 微球中的抗原在 T 淋巴细胞测定中诱导更强的白细胞介素-2 分泌。用 PLGA-MDP-TB10.4-Ag85B 脉冲的巨噬细胞表现出延长的表位呈递。
可吸入尺寸的 PLGA 微球能够以免疫相关的方式有效递呈重组 TB10.4-Ag85B 至巨噬细胞。这些结果为进一步研究 PLGA 颗粒在肺部递送来预防结核分枝杆菌感染的疫苗的潜在用途奠定了基础。
