BIOCHEMICAL CYTOLOGY OF HOST-PARASITE INTERACTIONS IN PARASITIC PROTOZOA

利什曼原虫和锥虫的细胞生物学和生物化学是 作为细胞内和细胞外寄生的模型， 分别 由于宿主和寄生虫之间的所有相互作用都发生在 水平的寄生虫表面膜（SM），重点放在：1）其 综合生化表征和2）定义其在 寄生虫生存 用Western印迹法检测了L. donovani到哪个内脏 利什曼病患者作出IgG应答。 糖脂 L.的组成部分。donovani前鞭毛体SM。 分泌的 对前鞭毛体酸性磷酸酶进行了纯化、化学表征， 无鞭毛体也会释放这种抗体， 酶和内脏患者产生针对它的IgG抗体。 前鞭毛体被激活，并被杀死的替代 补体途径和C3结合的主要靶点被鉴定为 SM蛋白的分子量为140 KD。 核糖的主动转运系统是 在L. donovani promastigotes. 调控 SM 3 '-核苷酸酶活性在诱导型遗传条件下， 控制 一种驱动膜运输过程的SM质子-ATP酶， 鉴定、细胞化学定位和酶学表征。 的 通过PCR鉴定了前鞭毛体的SM葡萄糖转运蛋白， 光亲和标记和部分表征。 基因组克隆编码 寄生虫SM抗原的鉴定，分离和表征， 使用抗SM血清和来自内脏的血清的表达载体系统 利什曼病患者。

The cell biology and biochemistry of Leishmania and Trypanosoma are investigated as models of intra- and extracellular parasitism, respectively. As all interactions between host and parasite occur at the level of the parasite surface membrane (SM), emphasis is placed on: 1) its integrated biochemical characterization and 2) defining its roles in parasite survival. Using Western blots, the major SM antigens of L. donovani to which visceral leishmaniasis patients make IgG responses were identified. The glycolipid constitutents of L. donovani promastigote SM were delineated. A secreted promastigote acid phosphatase was purified, chemically characterized and monoclonal antibodies raised against it. Amastigotes also release this enzyme and visceral patients make IgG antibodies against it. Log-phase promastigotes were shown to activate and be killed by the alternate complement pathway and the primary target of C3-binding was identified as a SM protein of 140 Kd. An active transport system for ribose was demonstrated and characterized in L. donovani promastigotes. Regulation of SM 3'-nucleotidase activity was shown to be under inducible genetic control. A SM proton-ATPase which drives membrane transport processes was identified, cytochemically localized and enzymatically characterized. The SM glucose transport protein of promastigotes was identified via photoaffinity labeling and partially characterized. Genomic clones coding for parasite SM antigens were identified, isolated and characterized from an expression-vector system using both anti-SM sera and sera from visceral leishmaniasis patients.