Cameron C E, Castro C, Lukehart S A, Van Voorhis W C
Department of Medicine, University of Washington, Seattle, Washington, USA.
Infect Immun. 1998 Dec;66(12):5763-70. doi: 10.1128/IAI.66.12.5763-5770.1998.
Infectious syphilis, caused by the spirochete bacterium Treponema pallidum subsp. pallidum, remains a public health concern worldwide. The immune-response evasion mechanisms employed by T. pallidum are poorly understood, and prior attempts to identify immunoprotective antigens for subsequent vaccine design have been unsuccessful. Previous investigations conducted in our laboratory identified the T. pallidum glycerophosphodiester phosphodiesterase as a potential immunoprotective antigen by using a differential immunologic expression library screen. In studies reported here, heterologous expression of the T. pallidum glycerophosphodiester phosphodiesterase in Escherichia coli yielded a full-length, enzymatically active protein. Characterization of the recombinant molecule showed it to be bifunctional, in that it exhibited specific binding to human immunoglobulin A (IgA), IgD, and IgG in addition to possessing enzymatic activity. IgG fractionation studies revealed specific binding of the recombinant enzyme to the Fc fragment of human IgG, a characteristic that may play a role in enabling the syphilis spirochete to evade the host immune response. In further investigations, immunization with the recombinant enzyme significantly protected rabbits from subsequent T. pallidum challenge, altering lesion development at the sites of challenge. In all cases, animals immunized with the recombinant molecule developed atypical pale, flat, slightly indurated, and nonulcerative reactions at the challenge sites that resolved before lesions appeared in the control animals. Although protection in the immunized rabbits was incomplete, as demonstrated by the presence of T. pallidum in the rabbit infectivity test, glycerophosphodiester phosphodiesterase nevertheless represents a significantly immunoprotective T. pallidum antigen and thus may be useful for inclusion in an antigen cocktail vaccine for syphilis.
由梅毒螺旋体苍白亚种引起的感染性梅毒仍是全球公共卫生关注的问题。梅毒螺旋体所采用的免疫反应逃避机制尚不清楚,此前为后续疫苗设计鉴定免疫保护性抗原的尝试均未成功。我们实验室先前进行的研究通过差异免疫表达文库筛选,将梅毒螺旋体甘油磷酸二酯磷酸二酯酶鉴定为一种潜在的免疫保护性抗原。在本文报道的研究中,梅毒螺旋体甘油磷酸二酯磷酸二酯酶在大肠杆菌中的异源表达产生了一种全长的、具有酶活性的蛋白质。对该重组分子的表征表明它具有双功能,因为它除了具有酶活性外,还表现出与人免疫球蛋白A(IgA)、IgD和IgG的特异性结合。IgG分级分离研究揭示了重组酶与人IgG的Fc片段的特异性结合,这一特性可能在使梅毒螺旋体逃避宿主免疫反应中发挥作用。在进一步的研究中,用重组酶免疫可显著保护兔子免受后续梅毒螺旋体攻击,改变攻击部位的病变发展。在所有情况下,用重组分子免疫的动物在攻击部位出现非典型的苍白、扁平、轻度硬结和非溃疡性反应,这些反应在对照动物出现病变之前就已消退。尽管在兔感染性试验中证明免疫兔子的保护作用不完全,但甘油磷酸二酯磷酸二酯酶仍然是一种具有显著免疫保护作用的梅毒螺旋体抗原,因此可能有助于纳入梅毒抗原混合疫苗中。