Robson K J, Frevert U, Reckmann I, Cowan G, Beier J, Scragg I G, Takehara K, Bishop D H, Pradel G, Sinden R
MRC Molecular Haematology Unit, John Radcliffe Hospital, Headington, Oxford, UK.
EMBO J. 1995 Aug 15;14(16):3883-94. doi: 10.1002/j.1460-2075.1995.tb00060.x.
Plasmodium sporozoites collected from oocysts, haemocoel and salivary glands of the mosquito show profound differences in their biological properties such as motility, ability to induce protective immune response and infectivity for vertebrate host cells. Sporozoites from salivary glands are much more infectious than those from oocysts and haemocoel. Differential expression of proteins, such as the circumsporozoite (CS) protein and the thrombospondin-related adhesive protein (TRAP), implicated in sporozoite recognition and entry into hepatocytes may account for the development of infectivity during ontogeny. We have carried out a series of experiments to: (i) analyse the expression and localization of TRAP in P.falciparum sporozoites during development in the mosquito; and (ii) elucidate the biochemical and adhesive properties of recombinant TRAP. Our data indicate that TRAP is not expressed in oocysts, whereas variable amounts of CS protein are found in this parasite developmental stage. Hemocoel sporozoites display the distinct phenotypes TRAP- CS protein+ and TRAP+ CS protein+ at a frequency of 98.5 and 1.5% respectively. Salivary gland sporozoites are all TRAP+ CS protein+. We also provide experimental evidence showing that recombinant TRAP binds to the basolateral cell membrane of hepatocytes in the Disse's space and that sulfated glycoconjugates function as TRAP ligands on human hepatocytes.
从按蚊的卵囊、血腔和唾液腺中收集的疟原虫子孢子在其生物学特性方面表现出显著差异,如运动性、诱导保护性免疫反应的能力以及对脊椎动物宿主细胞的感染性。来自唾液腺的子孢子比来自卵囊和血腔的子孢子更具感染性。参与子孢子识别和进入肝细胞的蛋白质,如环子孢子蛋白(CS)和血小板反应蛋白相关黏附蛋白(TRAP)的差异表达,可能是子孢子个体发育过程中感染性形成的原因。我们进行了一系列实验,以:(i)分析疟原虫子孢子在按蚊体内发育过程中TRAP的表达和定位;(ii)阐明重组TRAP的生化和黏附特性。我们的数据表明,TRAP在卵囊中不表达,而在此寄生虫发育阶段可发现不同量的CS蛋白。血腔子孢子分别以98.5%和1.5%的频率表现出TRAP - CS蛋白+和TRAP+ CS蛋白+这两种不同的表型。唾液腺子孢子均为TRAP+ CS蛋白+。我们还提供了实验证据,表明重组TRAP与狄氏间隙中肝细胞的基底外侧细胞膜结合,并且硫酸化糖缀合物在人肝细胞上作为TRAP配体发挥作用。