Wrightsman R A, Leon W, Manning J E
Infect Immun. 1986 Aug;53(2):235-9. doi: 10.1128/iai.53.2.235-239.1986.
Monoclonal antibodies reactive with the surface antigens of the Peru strain of Trypanosoma cruzi were analyzed by Western blots and immunofluorescence assays to determine their reactivity with three life cycle stages and five strain isolates of T. cruzi. One monoclonal antibody, 7.6, recognized a 68-kilodalton (kDa) polypeptide in Western blots of Peru strain trypomastigotes, epimastigotes, and amastigotes. A 68-kDa polypeptide was also detected by monoclonal antibody 7.6 in trypomastigotes of the CL and Y strains and in the clonal isolates Esmeraldo clone 3 and Silvio X10 clone 1. Positive immunofluorescence results were obtained for all life cycle stages of the five strains that were reacted with monoclonal antibody 7.6, thus indicating that the antigen recognized by monoclonal antibody 7.6 is universally present in all T. cruzi strains tested. In contrast, monoclonal antibody 4.2 reacted with a polypeptide doublet of 90 and 105 kDa in Western blots of Peru strain trypomastigotes, but it did not detect these antigens in epimastigotes or amastigotes. The same polypeptide doublet of 90 and 105 kDa was also detected in Western blots of Y strain trypomastigotes; however, no bands were detected in blots of strain CL or isolate Silvio X10 clone 1 trypomastigotes. In blots of Esmeraldo clone 3 trypomastigotes, a single band of 130 kDa was detected by monoclonal antibody 4.2. In immunofluorescence assays of monoclonal antibody 4.2, positive reactions were obtained only with trypomastigotes of Peru, Y, and Esmeraldo clone 3 strains. Thus, monoclonal antibody 4.2 recognizes a trypomastigote-specific antigen which is not universally present on all strains of T. cruzi.
通过蛋白质免疫印迹法和免疫荧光测定法分析了与克氏锥虫秘鲁株表面抗原反应的单克隆抗体,以确定它们与克氏锥虫三个生命周期阶段和五个菌株分离株的反应性。一种单克隆抗体7.6在秘鲁株锥鞭毛体、上鞭毛体和无鞭毛体的蛋白质免疫印迹中识别出一条68千道尔顿(kDa)的多肽。单克隆抗体7.6在CL和Y株的锥鞭毛体以及克隆分离株埃斯梅拉尔多克隆3和西尔维奥X10克隆1中也检测到了一条68-kDa的多肽。用单克隆抗体7.6反应的五个菌株的所有生命周期阶段均获得了阳性免疫荧光结果,这表明单克隆抗体7.6识别的抗原普遍存在于所有测试的克氏锥虫菌株中。相比之下,单克隆抗体4.2在秘鲁株锥鞭毛体的蛋白质免疫印迹中与90和105 kDa的多肽 doublet 发生反应,但在上鞭毛体或无鞭毛体中未检测到这些抗原。在Y株锥鞭毛体的蛋白质免疫印迹中也检测到了相同的90和105 kDa的多肽 doublet;然而,在CL株或分离株西尔维奥X10克隆1锥鞭毛体的印迹中未检测到条带。在埃斯梅拉尔多克隆3锥鞭毛体的印迹中,单克隆抗体4.2检测到一条130 kDa的单带。在单克隆抗体4.2的免疫荧光测定中,仅在秘鲁、Y和埃斯梅拉尔多克隆3株的锥鞭毛体中获得了阳性反应。因此,单克隆抗体4.2识别一种锥鞭毛体特异性抗原,该抗原并非普遍存在于所有克氏锥虫菌株中。