STRUCTURAL DETERMINATION OF NEISSERIA GONORRHOEAE OLIGOSACCHARIDES FROM ISOLATES

滴虫寄生虫，阴道毛滴虫，导致一个主要的 性传播疾病，滴虫病，在人类，感染 具有重要的医学意义 T迷走神经是最常见的 女性阴道炎的原因，并已与几个主要的 病理症状，如严重的阴道炎症， 刺激，不孕，早产，低出生体重婴儿， 浸润性宫颈癌和子宫内传播增加 也易感染人类免疫缺陷病毒 HIV病毒感染。 寄生虫粘附于宿主的能力 细胞在建立感染中起着不可或缺的作用。 等 相互作用可由细胞表面糖缀合物介导。 的 该项目的长期目标是使用高度纯化的寄生虫 糖缀合物产物，以确定细胞结合的和游离的 脂磷酸聚糖（TV-LPG）在T迷走细胞感染病理学中的作用， 和免疫力。 寄生虫具有新的LPG样糖复合物（3 X 106拷贝/寄生虫）通过肌醇锚定在细胞表面 磷酸神经酰胺，不同于迄今报道的任何其他GPI锚。 实验结果提示TV-LPG参与了T细胞的粘附 通过特异性受体-配体对HeLa细胞进行迷走神经刺激 互动 纯化的水溶性TV-LPG相关聚糖（TV-GL） 也可能参与了T 迷走神经发病机制 该项目的具体目标是：1） 建立人阴道上皮细胞（HVEC）体外培养物 研究类LPG糖复合物在粘附中的作用 的T迷走细胞HVECs，并检查激素的影响， 粘附T迷走神经; II）结构表征TV LPG， 阐明其与水溶性TV-GL的关系; III）产生 抗LPG及其片段的单克隆抗体（mAb），以检查 所述LPG表位在活寄生虫的表面上表达; 和IV）确定是否特异于表面表达的LPG的mAb 表位抑制HVEC的粘附，并对寄生虫具有细胞毒性 体外 来自培养的寄生虫的糖缀合物通过以下分析： 化学和酶消化结合HPLC，Glyko-FACE， GC/M[S]、MS、MALDI-TOF MS和NMR。 HVECS的培养方式和我们 建立牛“阴道”上皮细胞，研究牛 滴虫的发病机理，并将其暴露于激素中进行研究， T迷走细胞与HVEC的结合。 这些生物和化学 研究将提供结构/功能的基本知识 TV-LPG的关系，并将导致更好地了解 宿主-寄生虫致病的分子机制 交互. 此外，这些发现将有助于 发展有效的诊断和治疗人类 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.