IMMUNOPATHOGENESIS OF LYMPHATIC FILARIASIS

在拟议的研究中，我们将检查免疫机制，通过 通过与丝虫抗原局部接触而激活的T淋巴细胞 负责启动局部淋巴损伤，也就是中枢 感染伍氏杆菌的患者的病理表现 班克罗夫蒂。受体在丝虫免疫病理学中的作用 活化的T细胞及其受体表面的表达 内皮细胞表面局部产生的细胞因子将是 评估过了。因为没有班氏丝虫病的动物模型， 将对流行地区的受感染人类进行研究。通过比较和 寄生虫引起的淋巴病理与感染者的对比 到完全无症状但同样严重感染的微丝虫病 个体特异性免疫黏附受体的作用 整合素和免疫球蛋白超家族在介导破坏性反应中的作用 班克罗夫蒂杆菌感染中的淋巴病理将被调查。 具体的实验目标是：1)将感染者分成2个阶段 用于体外免疫学检测的临床小组和执行 炎性浅表淋巴管的免疫组织化学染色 免疫学结果的活体相关性；2)用细胞定量 酶联免疫吸附试验检测患者外周血单个核细胞培养上清中细胞因子的能力 诱导内皮细胞表面MHC分子和黏附受体的表达 体外模型系统中的细胞。特定细胞因子的水平将是 与受体表达相关。受体阻滞剂的作用 被评估；3)在体外黏附试验中，检查 患者T细胞表面受体在介导与培养内皮细胞黏附中的作用 用重组细胞因子人工刺激的细胞 诱导已知黏附受体的最大表达；以及4)检测 T细胞与经分析前激活的内皮细胞的黏附 与患者细胞来源的细胞因子上清孵育，利用 内皮细胞和T细胞受体的最佳实验条件 在具体目标2和3中分别定义的表述。 单抗阻断细胞表面受体的效果将被检测。 作为潜在靶点的黏附受体和配体 用单抗、可溶性受体进行免疫治疗阻断 类似物，或封闭肽将被识别，打开一个可能的新的 疾病控制的方法。最重要的是，进一步完善 体外模型系统将导致筛选能力的确定 重组丝虫抗原，当它们可用时，用于其 免疫致病潜能，导致鉴定抗- 病理疫苗候选者。

In the proposed study we will examine the immunologic mechanisms by which T-lymphocytes activated by local contact with filarial antigen are responsible for initiating the local lymphatic damage that is the central pathological manifestation seen in individuals infected with Wuchereria bancrofti. The contributions to filarial immunopathology of receptors expressed on the surface of activated T-cells and of receptors induced by locally produced cytokines on the surface of endothelial cells will be assessed. Because there is no animal model of bancroftian filariasis, infected humans in an endemic area will be studied. By comparing and contrasting infected individuals with parasite induced lymphatic pathology to completely asymptomatic but equally heavily infected microfilaremic individuals, the role of specific immunologic adhesion receptors of the integrin and immunoglobulin superfamilies in mediating the destructive lymphatic pathology seen in W. bancrofti infections will be investigated. The specific experimental aims are: 1) to stage infected individuals into 2 clinical groups for in vitro immunological assays and to perform immunohistologic staining of inflamed superficial lymphatic vessels for in vivo correlation of the immunological findings; 2) to quantitate by a cell ELISA the ability of cytokines in patient derived PBMC culture supernatants to induce expression of MHC molecules and adhesion receptors on endothelial cells in an in vitro model system. Levels of specific cytokines will be correlated with receptor expression. The effect of receptor blockade will be assessed; 3) to examine, in an in vitro adhesion assay, the role of receptors on patient T-cells in mediating adherence to cultured endothelial cells that have been artificially stimulated with recombinant cytokines to induce maximal expression of known adhesion receptors; and 4) to examine T-cell adhesion to endothelial cells which have been activated by preassay incubation with patient cell derived cytokine supernatants, utilizing optimal experimental conditions for endothelial cell and T-cell receptor expression that had been defined separately in Specific Aims 2 and 3. Effects of mAb blockade of cell surface receptors will be examined. Adhesion receptors and ligands that are potential targets for immuno-therapeutic blockade with monoclonal antibodies, soluble receptor analogues, or blocking peptides will be identified, opening a possible new approach to disease control. Most importantly, further refinement of the in vitro model system will lead to the ability to screen defined recombinant filarial antigens, when they become available, for their immunopathogenic potential, leading to the identification of anti- pathology vaccine candidates.