Alderete J F, Newton E, Dennis C, Neale K A
Department of Obstetrics and Gynecology, University of Texas Health Science Center, San Antonio 78284-7758.
Genitourin Med. 1991 Dec;67(6):469-74. doi: 10.1136/sti.67.6.469.
Patients with trichomoniasis have serum antibody to numerous T. vaginalis cysteine proteinases, indicating that the proteinases are expressed in vivo. It was important, therefore, to examine for the presence of soluble trichomonad proteinases and/or antibody to the proteinases in the vagina of infected women.
Vaginal washes (VWs) from 20 women were examined for the presence of proteinases by electrophoresis using acrylamide co-polymerised with gelatin as the indicator system. Antibody to proteinases in VWs was detected by an immunoprecipitation assay involving protein A-bearing Staphylococcus aureus first coated with anti-human immunoglobulin G (IgG) antibody, which was then added to VWs. For VWs having soluble proteinases, the bacteria were used to determine whether immune complexes between antibody and proteinases were present. VWs without soluble proteinases were incubated with the anti-human IgG treated bacteria before adding to detergent extracts of T. vaginalis. Individual isolates from the patients examined in this study were also analysed by one- and two-dimensional electrophoresis for their proteinase content. Finally, VWs were from patients without any history of other sexually transmitted diseases (STDs) as well as from individuals having numerous other STDs, including yeast, group B streptococcus, chlamydia, and syphilis.
Approximately one-third of patients had soluble proteinases in the VWs; the remaining two-thirds (70%) of patients and normal women had no detectable proteinases in VWs. Half of the patients without soluble proteinases had IgG which, when bound to S. aureus, immunoprecipitated many proteinases from a detergent extract of T. vaginalis. All soluble proteinases and those precipitated from trichomonal extracts were inhibited by inhibitors of cysteine proteinases. Finally, patients having trichomoniasis in addition to numerous other STD agents, including yeast, group B streptococcus, chlamydia, and syphilis did not have soluble proteinases in VWs. Equally noteworthy, some patients with soluble proteinases in VWs did not have other detectable STD agents.
Proteinases were detected in the vagina of some patients with trichomoniasis, and in most cases the proteinases were complexed with IgG, which was precipitated by S. aureus. Patients without soluble proteinases in VWs also had antibody specifically to trichomonad proteinases, again demonstrating both the expression and immunogenic nature of the proteinases in vivo. The absence of soluble proteinases in normal women and in patients having other STD agents as well as the presence of proteinases in VWs of patients without other detectable STD pathogens reinforced the idea that the proteinases were of T. vaginalis parasite origin. The findings of this study indicate that proteinases may be important to the T. vaginalis-host interrelationship.
滴虫病患者血清中存在针对多种阴道毛滴虫半胱氨酸蛋白酶的抗体,这表明这些蛋白酶在体内有表达。因此,检测感染女性阴道中可溶性滴虫蛋白酶和/或蛋白酶抗体的存在具有重要意义。
采用与明胶共聚的丙烯酰胺作为指示系统,通过电泳检测20名女性的阴道冲洗液(VW)中蛋白酶的存在情况。通过免疫沉淀试验检测VW中蛋白酶的抗体,该试验首先用抗人免疫球蛋白G(IgG)抗体包被带有蛋白A的金黄色葡萄球菌,然后将其加入VW中。对于含有可溶性蛋白酶的VW,用这些细菌来确定抗体与蛋白酶之间是否存在免疫复合物。对于没有可溶性蛋白酶的VW,在加入阴道毛滴虫去污剂提取物之前,先与经抗人IgG处理的细菌孵育。本研究中检测的患者的单个分离株也通过一维和二维电泳分析其蛋白酶含量。最后,VW来自没有任何其他性传播疾病(STD)病史的患者以及患有多种其他STD的个体,包括酵母菌、B族链球菌、衣原体和梅毒。
约三分之一的患者VW中含有可溶性蛋白酶;其余三分之二(70%)的患者以及正常女性的VW中未检测到蛋白酶。一半没有可溶性蛋白酶的患者有IgG,当与金黄色葡萄球菌结合时,可从阴道毛滴虫去污剂提取物中免疫沉淀出多种蛋白酶。所有可溶性蛋白酶以及从滴虫提取物中沉淀出的蛋白酶均被半胱氨酸蛋白酶抑制剂抑制。最后,除了酵母菌、B族链球菌、衣原体和梅毒等多种其他STD病原体外还患有滴虫病的患者,其VW中没有可溶性蛋白酶。同样值得注意的是,一些VW中含有可溶性蛋白酶的患者没有其他可检测到的STD病原体。
在一些滴虫病患者的阴道中检测到了蛋白酶,在大多数情况下,这些蛋白酶与IgG形成复合物,可被金黄色葡萄球菌沉淀。VW中没有可溶性蛋白酶的患者也有针对滴虫蛋白酶的特异性抗体,这再次证明了蛋白酶在体内的表达和免疫原性。正常女性和患有其他STD病原体的患者VW中没有可溶性蛋白酶,而没有其他可检测到的STD病原体的患者VW中存在蛋白酶,这强化了蛋白酶源自阴道毛滴虫寄生虫的观点。本研究结果表明,蛋白酶可能对阴道毛滴虫与宿主的相互关系很重要。