Bruce M C, Carter R N, Nakamura K, Aikawa M, Carter R
Institute of Cell, Animal and Population Biology, University of Edinburgh, UK.
Mol Biochem Parasitol. 1994 May;65(1):11-22. doi: 10.1016/0166-6851(94)90111-2.
The temporal expression during gametogenesis and the cellular location of the sexual stage specific protein Pfs16, a putative integral membrane protein of Plasmodium falciparum, was investigated using two monoclonal antibodies, 2G7 and 93A3A2. Using sorbitol synchronised, in vitro gametocyte cultures along with immunofluorescence assays, the time at which Pfs16 is first expressed during gametogenesis has been estimated to 35 hours post merozoite invasion. By immunofluorescence assays on thin blood smears monoclonal antibodies specific for Pfs16 react strongly with the gametocyte and also with vesicles within the red blood cell cytoplasm, many of which connect with the gametocyte cell. Purification of parasitophorous vacuole membranes from mature and immature gametocytes and immunoelectron microscopy on gametocytes during gametogenesis have allowed us to locate Pfs16 to the parasitophorous vacuole membrane. During gametogenesis this membrane is shed along with the red blood cell membrane. Immunofluorescence assays and immunoelectron microscopy studies of emerged gametes indicate that in a minority of cases the parasitophorous vacuole membrane along with Pfs16 can be retained to some extent on the gamete surface.
利用两种单克隆抗体2G7和93A3A2,研究了恶性疟原虫假定的整合膜蛋白——性阶段特异性蛋白Pfs16在配子发生过程中的表达时间和细胞定位。使用经山梨醇同步化的体外配子体培养物以及免疫荧光测定法,已估计出Pfs16在配子发生过程中首次表达的时间为裂殖子入侵后35小时。通过对薄血涂片进行免疫荧光测定,针对Pfs16的单克隆抗体与配子体以及红细胞胞质内的小泡强烈反应,其中许多小泡与配子体细胞相连。从成熟和未成熟配子体中纯化寄生泡膜,并对配子发生过程中的配子体进行免疫电子显微镜观察,使我们能够将Pfs16定位到寄生泡膜上。在配子发生过程中,该膜与红细胞膜一起脱落。对逸出的配子进行免疫荧光测定和免疫电子显微镜研究表明,在少数情况下,寄生泡膜连同Pfs16可以在一定程度上保留在配子表面。