ANTIGENIC ANALYSIS OF SEXUAL STAGES OF MALARIA PARASITES

我们的目标是，首先，继续发展一个25 kDa的性阶段表面 抗原Pfs 25作为潜在的候选疫苗;其次，克隆基因 对于其他已知的传播阻断免疫的靶抗原; 第三，鉴定有性阶段寄生虫上的新靶抗原;最后， 确定疟疾受精的分子机制 寄生虫 此外，我们正在研究寄生虫葡萄糖-6-磷酸 脱氢酶（G6 PD）酶和基因及其在保护中的作用 G6 PD缺乏症引起的。 以前，我们克隆了编码Pfs 25的基因，Pfs 25是一种主要的候选抗原 传播阻断疫苗 pfs 25现在已经在 细菌、酵母、牛痘和腺病毒感染的哺乳动物细胞， 瞬时转染的COS细胞和稳定转染的CHO细胞。 数据 从用牛痘产生的Pfs 25免疫的小鼠和Aotus猴中， 令人鼓舞的：来自接种活的重组 牛痘阻止疟疾传播。 我们还发现， 用纯化的酵母产生的Pfs 25免疫的猴子也获得了 传输阻断免疫。 几种佐剂系统已经被 检查并发现明矾或DPT足够。 克隆其他目标 组织抗原一直是一个问题，尽管我们现在相信， 克隆、测序并在E. coli Pfs 40，一个潜在的靶点 基于免疫遗传学数据的抗原。 小配子（雄性）特异性 单克隆抗体已经开发出来，可能会提供我们需要的入口， 了解受精的分子机制， 疟原虫 最后，我们已经证明疟原虫表达G6 PD， 组成性地，并且独立于宿主的G6 PD状态。 我们有 克隆了恶性疟原虫G6 PD基因。

Our goals are, first, continue development of a 25 kDa sexual stage surface antigen, Pfs25, as a potential vaccine candidate; second, clone the genes for the other known target antigens of transmission blocking immunity; third, identify new target antigens on sexual stage parasites; and finally, define the molecular mechanisms involved in fertilization of malarial parasites. In addition, we are studying the parasite glucose-6-phosphate dehydrogenase (G6PD) enzyme and gene and its role in the protection afforded by G6PD deficiency in humans. Previously we had cloned the gene encoding Pfs25, a prime candidate antigen for a transmission blocking vaccine. Pfs25 has now been expressed in bacteria, yeast, vaccinia ad adenovirus infected mammalian cells, transiently transfected COS cells, and stably transfected CHO cells. Data from mice and Aotus monkeys immunized with vaccinia-produced Pfs25 are very encouraging: sera from mice and monkeys inoculated with live, recombinant vaccinia block transmission of malaria. We have also found that mice and monkeys immunized with purified yeast-produced Pfs25 also acquire transmission blocking immunity. Several adjuvant systems have been examined and alum or DPT has been found adequate. Cloning the other target tissue antigens has been a problem, although we now believe that we have cloned, sequenced, and expressed in E. coli Pfs40, a potential target antigen based on immunogenetic data. A microgamete (male) specific monoclonal antibody has been developed and may provide the entre we need to understand the molecular mechanisms involved in fertilization in the malaria parasite. Finally, we have demonstrated that the malaria parasite expressed G6PD constitutively, and independently of the G6PD status of the host. We have cloned the P. falciparum G6PD gene.