Margos G, Kurtenbach K, Posnett E, Barker G C, Matsuoka H, Paton M G, Sinden R E
Department of Biology, Imperial College of Science, Technology and Medicine, London, UK.
Parasite Immunol. 1995 Apr;17(4):167-76. doi: 10.1111/j.1365-3024.1995.tb00886.x.
A surface protein of Plasmodium berghei ookinetes, Pbs21, was expressed in a baculovirus-insect cell system in cell culture and in Heliothis virescens larvae. Groups of BALB/c mice received two intraperitoneal inoculations of either i) Tris-buffer or homogenized H. virescens larvae infected with wild-type baculovirus; ii) enriched, homogenized ookinetes, or iii) homogenized H. virescens larvae expressing recombinant Pbs21 (rPbs21). All animals immunized with ookinetes or with rPbs21 had high titres of antibodies (IgG isotype) that bound to native Pbs21. The large majority of antibodies in immune sera of both groups recognized the antigen under non-reducing but not under reducing conditions. The predominant IgG-subclasses in mice immunized with ookinetes was IgG1 and in mice immunized with rPbs21, the subclasses were IgG1 and IgG2a. Immunization with rPbs21 reduced the infectivity of P.berghei to mosquitoes by 91% compared to a 99% reduction following immunization with ookinetes. This preliminary data indicate that rPbs21 expressed in this eukaryotic system induces a transmission-blocking immunity, which is more effective than that achieved using rPbs21 expressed in Escherichia coli (Matsuoka et al. 1994).
伯氏疟原虫动合子的一种表面蛋白Pbs21,在杆状病毒-昆虫细胞系统的细胞培养物以及烟草天蛾幼虫中表达。将BALB/c小鼠分组,分别腹腔注射以下物质:i)Tris缓冲液或感染野生型杆状病毒的匀浆烟草天蛾幼虫;ii)富集的匀浆动合子,或iii)表达重组Pbs21(rPbs21)的匀浆烟草天蛾幼虫。所有用动合子或rPbs21免疫的动物都产生了高滴度的能与天然Pbs21结合的抗体(IgG同种型)。两组免疫血清中的绝大多数抗体在非还原条件下能识别抗原,但在还原条件下不能。用动合子免疫的小鼠中主要的IgG亚类是IgG1,而用rPbs21免疫的小鼠中,亚类是IgG1和IgG2a。与用动合子免疫后感染性降低99%相比,用rPbs21免疫使伯氏疟原虫对蚊子的感染性降低了91%。这些初步数据表明,在这种真核系统中表达的rPbs21能诱导传播阻断免疫,这比使用在大肠杆菌中表达的rPbs21所达到的效果更有效(Matsuoka等人,1994年)。