Center for NanoBiotechnology and Life Science Research (CNBR), Alabama State University, Montgomery, AL 36104, USA.
Nanotechnology. 2012 Aug 17;23(32):325101. doi: 10.1088/0957-4484/23/32/325101. Epub 2012 Jul 23.
Development of a Chlamydia trachomatis vaccine has been a formidable task partly because of an ineffective delivery system. Our laboratory has generated a recombinant peptide of C. trachomatis major outer membrane protein (MOMP) (rMOMP-187) and demonstrated that it induced at 20 μg ml(-1) maximal interleukin (IL)-6 and IL-12p40 Th1 cytokines in mouse J774 macrophages. In a continuous pursuit of a C. trachomatis effective vaccine-delivery system, we encapsulated rMOMP-187 in poly(d,l-lactic-co-glycolic acid) (PLGA, 85:15 PLA/PGA ratio) to serve as a nanovaccine candidate. Physiochemical characterizations were assessed by Fourier transform-infrared spectroscopy, atomic force microscopy, Zetasizer, Zeta potential, transmission electron microcopy and differential scanning calorimetry. The encapsulated rMOMP-187 was small (∼200 nm) with an apparently smooth uniform oval structure, thermally stable (54 °C), negatively charged ( - 27.00 mV) and exhibited minimal toxicity at concentrations <250 μg ml (-1) to eukaryotic cells (>95% viable cells) over a 24-72 h period. We achieved a high encapsulation efficiency of rMOMP-187 (∼98%) in PLGA, a loading peptide capacity of 2.7% and a slow release of the encapsulated peptide. Stimulation of J774 macrophages with a concentration as low as 1 μg ml (-1) of encapsulated rMOMP-187 evoked high production levels of the Th1 cytokines IL-6 (874 pg ml(-1)) and IL-12p40 (674 pg ml(-1)) as well as nitric oxide (8 μM) at 24 h post-stimulation, and in a dose-response and time-kinetics manner. Our data indicate the successful encapsulation and characterization of rMOMP-187 in PLGA and, more importantly, that PLGA enhanced the capacity of the peptide to induce Th1 cytokines and NO in vitro. These findings make this nanovaccine an attractive candidate in pursuit of an efficacious vaccine against C. trachomatis.
沙眼衣原体疫苗的研发一直是一项艰巨的任务,部分原因是缺乏有效的递送系统。我们的实验室已经生成了沙眼衣原体主要外膜蛋白(MOMP)的重组肽(rMOMP-187),并证明它在 20 μg ml(-1)时在小鼠 J774 巨噬细胞中诱导了最大的白细胞介素(IL)-6 和 IL-12p40 Th1 细胞因子。为了不断寻求有效的沙眼衣原体疫苗递送系统,我们将 rMOMP-187 包裹在聚(D,L-乳酸-共-乙醇酸)(PLGA,85:15 PLA/PGA 比)中,作为纳米疫苗候选物。通过傅里叶变换红外光谱、原子力显微镜、Zetasizer、Zeta 电位、透射电子显微镜和差示扫描量热法评估物理化学性质。包裹的 rMOMP-187 较小(约 200nm),具有明显的光滑均匀的椭圆形结构,热稳定性(54°C),带负电荷(-27.00 mV),在 24-72 小时内浓度<250μg ml(-1)时对真核细胞(>95%存活细胞)的毒性最小。我们在 PLGA 中实现了 rMOMP-187 的高包封效率(约 98%)、肽的载药量为 2.7%和包裹肽的缓慢释放。刺激 J774 巨噬细胞的浓度低至 1μg ml(-1)的包裹 rMOMP-187 在刺激后 24 小时可引发高水平的 Th1 细胞因子 IL-6(874 pg ml(-1))和 IL-12p40(674 pg ml(-1))以及一氧化氮(8 μM),呈剂量反应和时间动力学方式。我们的数据表明 rMOMP-187 在 PLGA 中的成功包封和表征,更重要的是,PLGA 增强了肽诱导 Th1 细胞因子和 NO 的能力。这些发现使这种纳米疫苗成为针对沙眼衣原体的有效疫苗的有吸引力的候选物。