MEROZOITE MONOCLONAL ANTIBODY CHARACTERIZATION AND IN VITRO C. PARVUM CULTURING

艾滋病患者中由于隐孢子虫引起的衰弱性感染是 可能是I型裂殖体继续不受控制地循环的结果， 自身感染孢子体阶段。 第一类对数乘法 裂殖子被认为是导致 这些病人的感染。 目前没有有效的 化疗药物可用于对抗这种感染。 证据 然而，出现了阶段特异性或交叉反应性， 抗体，包括单克隆抗体，可用于消除这种免疫缺陷。 感染. 这一年最后延续的主要目标是 因此，项目将完成确定和 表征来源于裂殖子的单个和合并的MAb， 然后可以传递到项目2并与MAb结合使用 针对子孢子阶段推导，以确定最佳的单克隆抗体库 对C.小的 实现这一目标的具体办法包括： 1)完成电流的表征和中和测试 和利用体外感染模型的新衍生的mAb，和2） 扩大选择裂殖子中和敏感的单克隆抗体生产 表位的体外生物反应器系统。 终点目标 是提供最有用的抗裂殖子单克隆抗体用于测试， 与抗子孢子单克隆抗体合并，其可以在Balb/c和 抗肠道隐孢子虫病的SCID小鼠模型。 追求这些 目标应有助于确定预防或 治疗C.艾滋病患者的细小病毒感染。

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.