Kan S C, Yamaga K M, Kramer K J, Case S E, Siddiqui W A
Infect Immun. 1984 Jan;43(1):276-82. doi: 10.1128/iai.43.1.276-282.1984.
Serum samples from Aotus trivirgatus subsp. griseimembra monkeys obtained at different stages of a vaccination experiment were analyzed for total antibody titer to Plasmodium falciparum and were used for identifying protective antigens of the human malaria parasite. Total malarial antibody titers were higher in serum samples from protected monkeys (vaccinated with antigen in an adjuvant) than in those from unprotected monkeys (vaccinated with either antigen or adjuvant only). Parasite proteins were labeled with [3H]isoleucine, solubilized with nonionic detergent, and reacted with immune Aotus sera. Immunoprecipitates obtained were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and fluorography. Thirteen protein antigen bands in the molecular weight range 73,000 to 180,000 were resolved. Serum samples obtained from protected Aotus monkeys reacted more intensely with these proteins than samples from unprotected monkeys did. Evidence is presented that the protective antigen is not a single, normally nonimmunogenic, protein that is recognized only in protected monkeys. Rather, the present data indicate that a heightened immune response to multiple proteins correlated with in vivo protection to P. falciparum in Aotus monkeys. This finding may have a significant bearing on strategies for the development of a human P. falciparum vaccine.
对在接种实验不同阶段获取的三带犰狳亚种灰膜犰狳猴的血清样本进行分析,以检测其对恶性疟原虫的总抗体滴度,并用于鉴定人类疟原虫的保护性抗原。受保护猴子(用佐剂中的抗原进行接种)血清样本中的总疟疾抗体滴度高于未受保护猴子(仅接种抗原或佐剂)的血清样本。用[3H]异亮氨酸标记寄生虫蛋白,用非离子去污剂使其溶解,并与免疫犰狳血清反应。对获得的免疫沉淀物进行十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和荧光自显影分析。分辨出分子量范围在73,000至180,000之间的13条蛋白质抗原带。与未受保护猴子的样本相比,从受保护的犰狳猴获得的血清样本与这些蛋白质的反应更强烈。有证据表明,保护性抗原不是单一的、通常无免疫原性的、仅在受保护猴子中被识别的蛋白质。相反,目前的数据表明,对多种蛋白质的免疫反应增强与犰狳猴体内对恶性疟原虫的保护相关。这一发现可能对人类恶性疟原虫疫苗的开发策略具有重要意义。