MEROZOITE MONOCLONAL ANTIBODY CHARACTERIZATION AND IN VITRO C. PARVUM CULTURING

艾滋病患者中由微小隐孢子虫引起的衰弱感染 很可能是继续无节制地回收第一类Meront和 自体感染孢子体阶段。第I类的对数乘法 裂殖子被认为是对 这些患者的压倒性感染。目前还没有有效的 可用来对抗这种感染的化疗药物。有证据表明 然而，出现了暗示特定阶段或交叉反应的 抗体，包括单克隆体，可能有助于消除这种 感染。这一年的最终延续的主要目标是 因此，该项目将完成确定和 鉴定来自裂殖子的单个和集合的单抗 然后可以传递给项目2并与单抗一起使用 对子孢子阶段进行衍生以确定单抗的最佳池 对微小隐孢子虫有中和作用。 为实现这一目标而采取的具体办法将包括； 1)完成电流的表征和中和试验 以及利用体外感染模型获得的新单抗和2) 中和敏感型裂殖子单抗的放大生产 利用体外生物反应器系统的表位。终点目标 是为了提供最有用的抗裂殖子单抗用于测试和 与抗子孢子单抗共用，可在Balb/c和 抗肠道隐孢子虫病的SCID小鼠模型。对这些的追求 目标应有助于确定预防或 治疗艾滋病患者的微小弧菌感染。

Debilitating infections due to Cryptosporidium parvum in AIDS patients are likely the result of the continued unchecked recycling of type I meront and autoinfective sporont stages. The logarithmic multiplication of type I merozoites has been cited as contributing most significantly to the overwhelming infection in these patients. There are presently no effective chemotherapeutic agents available to combat this infection. Evidence has emerged, however, to suggest that stage specific or cross reactive antibodies, including monoclonals, may be useful in abrogating such infections. The main objective of this one year final continuation project, therefore, will be to complete work on identifying and characterizing individual and pooled MAbs derived from merozoites which then can be passed on to Project 2 and used in conjunction with MAbs derived against the sporozoite stage to determine the optimal pool of MAbs having neutralization efficacy against C. parvum. Specific approaches to be taken to accomplish this objective will include; 1) completing the characterization and neutralization testing of current and newly derived mAbs utilizing in vitro models of infection and 2) scaling up MAb production of select merozoite neutralization-sensitive epitopes utilizing an in vitro bioreactor system. The endpoint objective is to provide the most useful of the anti-merozoite MAbs for testing and pooling with anti-sporozoite MAbs which can be tested in both Balb/c and SCID mouse models against intestinal cryptosporidiosis. Pursuit of these objectives should help define optimal strategies for preventing or treating C. parvum infections in AIDS patients.