CHOLERA: PATHOGENESIS AND IMMUNOLOGY

这项研究的目的是为全面了解 霍乱的发病机制和免疫学在生物体和 分子水平，并期望这将导致 发展合理有效的免疫预防手段。AS 霍乱是越来越多的肠道毒素的原型 肠道疾病，这些观察结果与更大的全球问题有关 分泌性腹泻病。很明显，霍乱这种疾病是 一种有效的免疫过程，向人类宿主呈现一种 由体内生长的霍乱弧菌精制的产品联合体。 这些因素包括：殖民因素(尚未确定)；表面成分 如脂多糖和外膜蛋白； 透明质酸/蛋白酶；其他酶；对甘露糖敏感(El Tor生物型)和 其他血凝素、鞭毛和肠毒素。这项建议 具体涉及：确定和表征 参与霍乱弧菌肠道黏附的因子(S) 上皮细胞；外膜抗原(特别是新的铁调节的 在体内表达的蛋白质)；以及抗原和抗体的定义 它可能是保护性的[包括功能结构域和表位 免疫优势，结合，霍乱原(B亚单位五聚体)部分 霍乱肠毒素]。技术包括：细菌结合研究； 蛋白质分离分析采用SDS-PAGE和免疫印迹； 涉及母乳蛋白质成分的微量抗菌检测， 多克隆和单克隆抗细菌抗体；以及抗毒素 霍乱肠毒素的单抗和合成肽 目标是开发一种合成疫苗。

The goals of this research are to contribute to the complete understanding of the pathogenesis and immunology of cholera at both the organismal and the molecular levels with the expectation that this will lead to the development of rational and effective means of immunoprophylaxis. As cholera is the prototype of an expanding number of enterotoxic enteropathies, the observations are pertinent to the larger global problem of secretory diarrheal disease. It is clear that the disease, cholera, is an effective immunizing process which presents to the human host a consortium of products elaborated by the cholera vibrios growing in vivo. These include: colonization factors (as yet undefined); surface components such as the lipopolysaccharide and outer membrane proteins; the HA/protease; other enzymes; the mannose-sensitive (El Tor biotype) and other hemagglutinins; the flagellum; and the enterotoxin. This proposal specifically addresses: the identification and characterization of factor(s) participating in adherence of Vibrio cholerae to intestinal epithelium; outer membrane antigens (particularly novel iron-regulated proteins expressed in vivo); and the definition of antigens and antibodies which may be protective [including functional domains and epitopes on the immunodominant, binding, choleragenoid (B-subunit pentamer) portion of the cholera enterotoxin]. Technology includes: bacterial binding studies; protein separation and analysis with SDS-PAGE and immunoblotting; micro-antibacterial assays involving protein components of mothers' milk, polyclonal and monoclonal antibacterial antibodies; and antitoxic monoclonal antibodies and synthetic peptides of the cholera enterotoxin with the objective of developing a synthetic vaccine.