Gonzalez-Ceron L, Rodriguez M H, Santillan F, Chavez B, Nettel J A, Hernandez-Avila J E, Kain K C
Centro de Investigación de Paludismo, Instituto Nacional de Salud Pública, 4 Norte and 19 Poniente, 30700 Tapachula, Chiapas, Mexico.
Exp Parasitol. 2001 Jul;98(3):152-61. doi: 10.1006/expr.2001.4626.
Anopheles albimanus and An. pseudopunctipennis differ in their susceptibilities to Plasmodium vivax circumsporozoite phenotypes. An. pseudopunctipennis is susceptible to phenotype VK247 but almost refractory to VK210. In contrast, An. albimanus is almost refractory to VK247 but susceptible to VK210. To investigate the site in the mosquito and the parasite stage at which resistance mechanisms affect VK247 development in An. albimanus, parasite development was followed in a series of experiments in which both mosquitoes species were simultaneously infected with blood from patients. Parasite phenotype was determined in mature oocysts and salivary gland sporozoites by use of immunofluorescence and Western blot assays and/or gene identification. Ookinete maturation and their densities within the bloodmeal bolus were similar in both mosquito species. Ookinete densities on the internal midgut surface of An. albimanus were 4.7 times higher than those in An. pseudopunctipennis; however, the densities of developing oocysts on the external midgut surface were 6.12 times higher in the latter species. Electron microscopy observation of ookinetes in An. albimanus midgut epithelium indicated severe parasite damage. These results indicate that P. vivax VK247 parasites are destroyed at different parasite stages during migration in An. albimanus midguts. A portion, accumulated on the internal midgut surface, is probably destroyed by the mosquito's digestive enzymes and another portion is most likely destroyed by mosquito defense molecules within the midgut epithelium. A third group, reaching the external midgut surface, initiates oocyst development, but over 90% of them interrupt their development and die. The identification of mechanisms that participate in parasite destruction could provide new elements to construct transgenic mosquitoes resistant to malaria parasites.
白纹按蚊和伪点按蚊对间日疟原虫环子孢子表型的易感性不同。伪点按蚊对VK247表型敏感,但对VK210几乎具有抗性。相比之下,白纹按蚊对VK247几乎具有抗性,但对VK210敏感。为了研究蚊子体内的作用位点以及抗性机制影响白纹按蚊中VK247发育的寄生虫阶段,在一系列实验中跟踪了寄生虫的发育情况,在这些实验中,两种蚊子同时感染了患者的血液。通过免疫荧光和蛋白质印迹分析和/或基因鉴定来确定成熟卵囊和唾液腺子孢子中的寄生虫表型。两种蚊子中动合子的成熟及其在血餐团中的密度相似。白纹按蚊中肠内表面的动合子密度比伪点按蚊高4.7倍;然而,后一种蚊子中肠外表面发育中的卵囊密度高6.12倍。对白纹按蚊中肠上皮中动合子的电子显微镜观察表明寄生虫受到严重损伤。这些结果表明,间日疟原虫VK247寄生虫在白纹按蚊中肠迁移过程中的不同寄生虫阶段被破坏。一部分积聚在中肠内表面,可能被蚊子的消化酶破坏,另一部分很可能被中肠上皮内的蚊子防御分子破坏。第三组到达中肠外表面,开始卵囊发育,但其中超过90%会中断发育并死亡。参与寄生虫破坏的机制的鉴定可以为构建抗疟原虫的转基因蚊子提供新的要素。