Center for Immunobiology and Vaccine Development, Children's Hospital Oakland Research Institute, Oakland, CA 94609, USA.
Biomaterials. 2012 Jul;33(20):5166-74. doi: 10.1016/j.biomaterials.2012.03.054. Epub 2012 Apr 9.
Developing materials that are effective against sexually transmitted pathogens such as Chlamydia trachomatis (Ct) and HIV-1 is challenging both in terms of material selection and improving bio-membrane and cellular permeability at desired mucosal sites. Here, we engineered the prokaryotic bacterial virus (M13 phage) carrying two functional peptides, integrin binding peptide (RGD) and a segment of the polymorphic membrane protein D (PmpD) from Ct, as a phage-based material that can ameliorate Ct infection. Ct is a globally prevalent human pathogen for which there are no effective vaccines or microbicides. We show that engineered phage stably express both RGD motifs and Ct peptides and traffic intracellularly and into the lumen of the inclusion in which the organism resides within the host cell. Engineered phage were able to significantly reduce Ct infection in both HeLa and primary endocervical cells compared with Ct infection alone. Polyclonal antibodies raised against PmpD and co-incubated with constructs prior to infection did not alter the course of infection, indicating that PmpD is responsible for the observed decrease in Ct infection. Our results suggest that phage-based design approaches to vector delivery that overcome mucosal cellular barriers may be effective in preventing Ct and other sexually transmitted pathogens.
开发针对沙眼衣原体(Ct)和 HIV-1 等性传播病原体的有效材料在材料选择以及提高所需粘膜部位的生物膜和细胞通透性方面都具有挑战性。在这里,我们构建了携带两种功能肽(整合素结合肽(RGD)和 Ct 多态膜蛋白 D(PmpD)片段)的原核细菌病毒(M13 噬菌体),作为一种基于噬菌体的材料,可以改善 Ct 感染。Ct 是一种在全球范围内流行的人类病原体,目前尚无有效的疫苗或杀微生物剂。我们表明,工程噬菌体稳定表达 RGD 基序和 Ct 肽,并在细胞内和包含体的腔室内运输,包含体内的生物体存在于宿主细胞内。与单独的 Ct 感染相比,工程噬菌体能够显著降低 HeLa 和原代宫颈内细胞中的 Ct 感染。与感染前与构建体共孵育的针对 PmpD 的多克隆抗体并未改变感染过程,表明 PmpD 是导致观察到的 Ct 感染减少的原因。我们的研究结果表明,克服粘膜细胞屏障的基于噬菌体的载体传递设计方法可能有效预防 Ct 和其他性传播病原体。