SURFACE ANTIGENS OF TREPONEMA PALLIDUM

梅毒由梅毒螺旋体引起，仍然是一个重大的公共卫生问题，并已被证明会加剧艾滋病毒的传播。需要新的梅毒预防手段，包括疫苗开发。为了合理地开发疫苗，必须根据抗原在发病机制和免疫方面的功能优先进行检测。抗体对梅毒螺旋体的调理作用对细菌清除和巨噬细胞介导的杀伤至关重要。因此，我们将从重组表达文库中筛选出与光学兔血清(ORS)反应的克隆。ORS是免疫的兔血清，从中吸附了许多“不相关的”抗体(那些对调理不重要的抗体，包括主要脂蛋白和内鞭毛的抗体)；吸附后保留了全部调理能力。选定的克隆将优先进行进一步检查，因为它们与非调理抗血清(抗疏螺旋体、抗钩端螺旋体)缺乏反应性。有希望的克隆将被测序，其完整的开放阅读框架(ORF)将在新发布的梅毒螺旋体基因组序列中确定。ORF将被表达为重组蛋白并产生抗体。这些抗体将被用来确定抗原是否位于梅毒螺旋体的表面。这些分子在梅毒螺旋体附着到真核细胞中的作用，这是发病的早期步骤，将通过测试抗体在体外阻止附着的能力来检验。这些抗原在调理中的作用将通过测试抗血清促进兔腹膜巨噬细胞吞噬梅毒螺旋体的能力来检验。根据其表面定位或在调理或细胞黏附中的作用，似乎有希望的抗原将被测试其保护兔免受梅毒螺旋体攻击的能力。这种基于已知在致病和免疫中的重要功能来选择感兴趣的抗原的集中方法，将识别梅毒螺旋体的重要毒力因素，并将识别潜在的疫苗候选进行进一步研究。

Syphilis, caused by Treponema pallidum, continues to be a significant public health problem and has been demonstrated to augment the spread of HIV. New means of prevention of syphilis, including vaccine development, are needed. In order to approach vaccine development in a rational way, antigens must be prioritized for examination based upon their functions in pathogenesis and immunity. The opsonization of T. pallidum by antibodies is crucial for bacterial clearance and for macrophage-mediated killing. Therefore, a recombinant expression library will be screened for clones reacting with opsonic rabbit serum (ORS). The ORS is immune rabbit serum from which many "irrelevant" antibodies (those not important for opsonization, including antibodies to the major lipoproteins and the endoflagella) have been adsorbed; full opsonic capacity is retained following adsorption. Selected clones will be prioritized for further examination based upon their lack of reactivity with non-opsonic antisera (anti-Borrelia, anti-Leptospira). Promising clones will be sequenced and their full open reading frames (ORFs) will be identified in the newly released T. pallidum genome sequence. The ORFs will be expressed as recombinant proteins and antibodies will be produced. These antibodies will be used to determine if the antigens are located on the surface of T. pallidum. The role of these molecules in attachment of T. pallidum to eucaryotic cells, an early step in pathogenesis, will be examined by testing the ability of the antibodies to block adherence in vitro. The role of the antigens in opsonization will be examined by testing the antisera for their ability to facilitate phagocytosis of T. pallidum by rabbit peritoneal macrophages. Antigens that appear to be promising, based upon in their surface localization or their role in opsonization or cytoadherence, will be tested for their ability to protect rabbits from challenge with T. pallidum. This focused approach for selecting antigens of interest, based upon functions known to be important in pathogenesis and immunity, will identify important virulence factors of T. pallidum and will identify potential vaccine candidates for further study.