Ibañez C F, Affranchino J L, Frasch A C
Instituto de Investigaciones Bioquímicas Fundación Campomar, Buenos Aires, Argentina.
Mol Biochem Parasitol. 1987 Sep;25(2):175-84. doi: 10.1016/0166-6851(87)90006-5.
A genomic DNA library from Trypanosoma cruzi, the agent of Chagas' disease, was constructed in the gt11 lambda vector and was screened with serum from a Chagasic patient. Out of 53 positive clones, 23 plaques were purified to homogeneity and 10 different groups were defined by cross-hybridization experiments and by reaction of antibodies selected with products from each recombinant clone. Native T. cruzi proteins of molecular mass ranging from 85 to larger than 205 kDa that share antigenic determinants with products of the recombinant clones were observed in Western blots of parasite extracts. Some of the native proteins were detected in the trypomastigote stage of the parasite, while others were present in epimastigotes as well. The latter result was confirmed for some recombinant clones by hybridization of the cloned DNA with Northern blots of parasite RNA. Clones from each group reacted differently with nine sera from rabbits infected with several T. cruzi strains as well as with eight sera from human patients. Clone 7 was detected by all rabbit sera but not by three human sera. Conversely, clones 1, 2 and 30 were detected by all human sera but failed to be detected by most rabbit sera. We conclude that several proteins from T. cruzi are antigenically active during infection and that some of them differ in their ability to generate antibodies in rabbit or human infections.
用gt11 λ载体构建了来自恰加斯病病原体克氏锥虫的基因组DNA文库，并用一名恰加斯病患者的血清进行筛选。在53个阳性克隆中，23个噬菌斑被纯化至同质，通过交叉杂交实验以及用来自每个重组克隆的产物选择的抗体反应确定了10个不同的组。在寄生虫提取物的蛋白质印迹中观察到分子量范围从85 kDa到大于205 kDa的天然克氏锥虫蛋白，它们与重组克隆的产物共享抗原决定簇。一些天然蛋白在寄生虫的无鞭毛体阶段被检测到，而其他一些蛋白也存在于前鞭毛体中。通过将克隆的DNA与寄生虫RNA的Northern印迹杂交，对一些重组克隆证实了后一结果。来自每组的克隆与感染几种克氏锥虫菌株的兔子的九份血清以及八份人类患者血清的反应不同。克隆7被所有兔子血清检测到，但未被三份人类血清检测到。相反，克隆1、2和30被所有人类血清检测到，但大多数兔子血清未能检测到。我们得出结论，克氏锥虫的几种蛋白质在感染期间具有抗原活性，并且其中一些蛋白质在兔或人类感染中产生抗体的能力有所不同。
