沙眼衣原体是世界上最常见的细菌性性传播病原体。由于衣原体感染在很大程度上是无症状的,有可能出现严重的并发症,预防性疫苗可能是应对这种公共卫生威胁的最可行的长期解决方案。无细胞蛋白质合成(CFPS)利用细胞蛋白质制造机制与维持细胞活力的要求脱钩,提供灵活的潜力,快速,和去集中生产重组蛋白疫苗抗原。这里,我们使用CFPS产生假定的衣原体型三分泌系统(T3SS)针尖蛋白,CT584,用作小鼠模型中的疫苗抗原。高速原子力显微镜(HS-AFM)成像和计算机模拟证实,CFPS产生的CT584在免疫之前保留了天然样结构。雌性小鼠鼻内(i.n.)或肌内(i.m.)用CpG-1826MontanideISA720佐剂化的CT584引发,四周后,在104个衣原体包涵体形成单位(IFU)进行呼吸攻击之前,进行静脉注射。用CT584免疫产生了强大的抗体反应,但细胞介导的免疫力较弱,并且无法抵御如体重减轻所证明的静脉内攻击。肺部重量增加,肺部存在大量IFU。虽然单独的CT584可能不是理想的候选疫苗,CFPS可用于生产其他潜在衣原体抗原的速度和灵活性使其成为一种有吸引力的抗原生产技术。
Chlamydia trachomatis is the most prevalent bacterial sexually transmitted pathogen worldwide. Since chlamydial infection is largely asymptomatic with the potential for serious complications, a preventative vaccine is likely the most viable long-term answer to this public health threat. Cell-free protein synthesis (CFPS) utilizes the cellular protein manufacturing machinery decoupled from the requirement for maintaining cellular viability, offering the potential for flexible, rapid, and de-centralized production of recombinant protein vaccine antigens. Here, we use CFPS to produce the putative chlamydial type three secretion system (T3SS) needle-tip protein,
CT584, for use as a vaccine antigen in mouse models. High-speed atomic force microscopy (HS-AFM) imaging and computer simulations confirm that CFPS-produced
CT584 retains a native-like structure prior to immunization. Female mice were primed with
CT584 adjuvanted with CpG-1826 intranasally (i.n.) or CpG-1826 + Montanide ISA 720 intramuscularly (i.m.), followed four-weeks later by an i.m. boost before respiratory challenge with 104 inclusion forming units (IFU) of Chlamydia muridarum. Immunization with
CT584 generated robust antibody responses but weak cell mediated immunity and failed to protect against i.n. challenge as demonstrated by body weight loss, increased lungs\' weights and the presence of high numbers of IFUs in the lungs. While
CT584 alone may not be the ideal vaccine candidate, the speed and flexibility with which CFPS can be used to produce other potential chlamydial antigens makes it an attractive technique for antigen production.