In vitro assessment of halobacterial gas vesicles as a Chlamydia vaccine display and delivery system
In vitro assessment of halobacterial gas vesicles as a Chlamydia vaccine display and delivery system
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DOI：
10.1016/j.vaccine.2012.07.038
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发表时间：
2012-09-07
期刊：
影响因子：
5.5
通讯作者：
Webley, Wilmore C.
中科院分区：
文献类型：
--
作者：
Childs, Tawanna S.;Webley, Wilmore C.
沙眼衣原体是世界范围内细菌性传播疾病的主要原因，虽然抗生素治疗可以有效消除病原体，但高达70%的感染是无症状的。尽管在过去20年中持续努力，但有效的衣原体疫苗仍然难以捉摸，这在很大程度上是由于缺乏有效的递送系统。我们探索了使用来自盐生盐杆菌的气泡作为疫苗感兴趣的衣原体抗原的潜在展示和递送载体。将编码主要外膜蛋白（MOMP）、外膜复合物B（Omc B）和多态性外膜蛋白D（PompD）的各种大小的基因片段整合到这些稳定结构的表面上，并表达为气泡蛋白C（gvpC）的一部分。通过使用抗gvpC和抗衣原体抗体以及来自衣原体阳性患者的血清探测的蛋白质印迹证实重组蛋白的存在。组织培养评价显示重组气囊泡（r-Gv）在人和动物细胞系中的稳定性和时间依赖性降解。使用人包皮成纤维细胞（HFF）的体外评估证实了野生型和r-Gv与Toll样受体（TLR）4和5的接合，导致MyD 88活化、TNF-α、IL-6和IL-12产生。这些数据表明，r-GV可能是一种有效的，自然佐剂，时间释放抗原递送系统的免疫相关的衣原体疫苗抗原，很容易识别的人免疫血清。(c)2012爱思唯尔有限公司保留所有权利。
Chlamydia trachomatis is the leading cause of bacterial sexually transmitted disease worldwide and while antibiotic treatment is effective in eliminating the pathogen, up to 70% of all infections are asymptomatic. Despite sustained efforts over the past 2 decades, an effective chlamydial vaccine remains elusive, due in large part to the lack of an effective delivery system. We explored the use of gas vesicles derived from Halobacterium salinarium as a potential display and delivery vehicle for chlamydial antigens of vaccine interest. Various size gene fragments coding for the major outer membrane protein (MOMP), outer membrane complex B (OmcB) and polymorphic outer membrane protein D (PompD) were integrated into and expressed as part of the gas vesicle protein C (gvpC) on the surface of these stable structures. The presence of the recombinant proteins was confirmed by Western blots probed using anti-gvpC and anti-Chlamydia antibodies as well as sera from Chlamydia-positive patients. Tissue culture evaluation revealed stability and a time-dependent degradation of recombinant gas vesicles (r-Gv) in human and animal cell lines. In vitro assessment using human foreskin fibroblasts (HFF) confirmed Toll-like receptor (TLR) 4 and 5 engagement by wild type and r-Gv, leading to MyD88 activation, TNF-alpha, IL-6 and IL-12 production. The data suggest that r-GV could be an effective, naturally adjuvanting, time-release antigen delivery system for immunologically relevant Chlamydia vaccine antigens which are readily recognized by human immune sera. (c) 2012 Elsevier Ltd. All rights reserved.