STRUCTURAL DETERMINATION OF NEISSERIA GONORRHOEAE OLIGOSACCHARIDES FROM ISOLATES

滴虫寄生虫，阴道毛滴虫，引起一种主要的 性传播疾病，人类滴虫病，一种感染 具有重要的医学价值。阴道毛滴虫是最常见的 女性阴道炎的原因，并与几个主要的 病理症状如严重的阴道炎和 刺激，不孕不育，早产，低出生体重婴儿， 浸润性宫颈癌与宫内传播增加 细胞负性病毒与人类免疫缺陷易感性 病毒(HIV)感染。寄生虫附着在宿主上的能力 细胞在确定感染的过程中起着不可或缺的作用。是这样的 相互作用可能是由细胞表面的糖偶联物介导的。这个 该项目的长期目标是使用高度纯化的寄生虫 确定糖共轭产物对细胞结合和游离的作用 脂磷脂多糖(TV-LPG)对阴道毛滴虫感染病理的影响 和免疫。该寄生虫具有新的LPG样糖结合物(3 X 106拷贝/寄生虫)通过肌醇锚定在细胞表面 磷酸肌酰胺，有别于迄今报道的任何其他GPI锚定。 实验数据表明，TV-LPG参与了T细胞的黏附 阴道病毒通过一种特异性受体配体对HeLa细胞的作用 互动。提纯的TV-LPG相关多糖(TV-GL) 阴道毛滴虫的条件培养液也可能参与T 阴道病的发病机制。该项目的具体目标是：1) 人阴道上皮细胞体外培养方法的建立 类LPG糖偶联物参与粘连的研究 阴道毛滴虫对人血管内皮细胞的作用，并检测 阴道毛滴虫粘连；ii)电视液化石油气的结构特征和 阐明其与水溶性TV-GL的关系；iii)生产 抗LPG及其片段的单抗(MAbs) 活体寄生虫表面表达哪些LPG表位； 以及IV)以确定表面表达的LPG是否具有特异性单抗 表位抑制与人血管内皮细胞的黏附，对寄生虫有细胞毒性 在试管中。从培养的寄生虫中提取的糖偶联物通过 化学和酶消化结合高效液相色谱，糖脸， 石丹参[S，MS，MALDI-TOF MS和核磁共振]。HECs正在像我们一样培养 为牛“阴道”上皮细胞建立的研究牛 滴虫的致病机制，并将激素暴露研究 阴道毛滴虫与血管内皮细胞的结合。这些生物和化学物质 研究将提供有关结构/功能的基本知识 电视与液化石油气的关系，将有助于更好地理解 寄主-寄生虫致病的分子机制 互动。此外，这些发现将有助于 人类有效诊断和治疗方法的发展 滴虫病。

The trichomonad parasite, Trichomonas vaginalis, causes a major sexually transmitted disease, trichomoniasis, in humans, an infection of substantial medical importance. T vaginalis is the most common cause of vaginitis in women and has been linked to several major pathological symptoms such as severe vaginal inflammation and irritation, infertility, preterm delivery, low-birth-weight infants, invasive cervical carcinoma and increased intrauterine transmission of cytornegalovirus and also susceptibility to human immunodeficiency virus (HIV) infection. The ability of parasites to adhere to host cells plays an integral role in establishing infections. Such interactions may be mediated by cell surface glycoconjugates. The long term objective of this project is to use highly purified parasite glycoconjugate products to determine the role of cell-bound and free lipophosphoglycan (TV-LPG) upon T vaginalis infection pathology, andimmunity. The parasite possesses novel LPG-like glycoconjugates (3 X 106 copies/parasite) anchored on the cell surface via an inositol phosphoceramide, distinct from any other GPI anchor reported so far. The experimental data suggest that TV-LPG is involved in adhesion of T vaginalis to HeLa cells through a specific receptor-ligand interaction. The water soluble TV-LPG related glycan (TV-GL) purified from conditioned media of T vaginalis may also be involved in T vaginalis pathogenesis. The specific aims of the project are: 1) to establish in vitro cultures of human vaginal epithelial cells (HVECs) to study the involvement of the LPG-like glycoconjugates in adhesion of T vaginalis to HVECs and also examine the hormonal effects of adhesion of T vaginalis; II) to structurally characterize TV LPG and clarify its relationship to the water soluble TV-GL; III) to produce monoclonal antibodies (mAbs) against LPG and its fragments, to examine which LPG epitopes are expressed on the surface of the live parasite; and IV) to determine if mAbs specific for surface expressed LPG epitopes inhibit adhesion to HVECs and are cytotoxic to the parasite in vitro. The glycoconjugates from cultured parasites are analyzed by chemical and enzymatic digestion in combination with HPLC, Glyko-FACE, GC/M[S, MS, MALDI-TOF MS andNMR. HVECS are being cultured as we have established for bovine "vaginal' epithelial cells to study bovine trichomonad pathogenesis, and will be exposed to hormone to study the binding of T vaginalis to HVECs. These biological and chemical studies will provide essential knowledge of the structure/function relationships of TV-LPG and will lead to a better understanding of the molecular mechanisms of pathogenesis involved in host-parasite interactions. In addition, these findings will contribute to the development of effective diagnosis and therapies of human trichomoniasis.