Inoue N, Lluz A T, Mori T, Nagasawa H, Fujisak K, Mikami T
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Hokkaido, Japan.
J Vet Med Sci. 2000 Oct;62(10):1041-5. doi: 10.1292/jvms.62.1041.
Seven monoclonal antibodies (mAbs) were raised against Trypanosoma congolense procyclic form (PCF). Localization of the antigens recognized by the mAbs was determined in bloodstream form (BSF), PCF, epimastigote form (EMF) and metacyclic form (MCF) by confocal laser scanning microscopy (CLSM). Two mAbs (10F9 and 20H12) showed different fluorescent patterns among different life-cycle stages of the parasite. The 10F9 recognized a 76 kDa antigen of all life-cycle stages of the parasite and the antigen localization corresponded with that of a mitochondrion. While the 20H12 recognized 119 and 122 kDa antigens of all the life-cycle stages and the antigen localization corresponded with a flagellum in BSF and MCF, tip of a flagellum in PCF, and part of cytoplasm in EMF. Moreover, the 20H12 did not react to T. brucei gambiense, T. b. rhodesiense and T. evansi antigens in both CLSM and immunoblotting. Therefore, the antigens recognized by the 20H12 seem to be T. congolense specific. Although, further studies will be required for a full characterization of the T. congolense specific 119 and 122 kDa antigens, the mAb 20H12 and the specific antigens may be useful in not only establishment of T. congolense specific diagnosis methods but also studies on molecular mechanisms regulating differentiation of the parasite during life-cycle.
制备了七种针对刚果锥虫前循环型（PCF）的单克隆抗体（mAb）。通过共聚焦激光扫描显微镜（CLSM）确定了mAb识别的抗原在血流型（BSF）、PCF、上鞭毛体型（EMF）和循环后期型（MCF）中的定位。两种mAb（10F9和20H12）在寄生虫的不同生命周期阶段表现出不同的荧光模式。10F9识别寄生虫所有生命周期阶段的一种76 kDa抗原，且抗原定位与线粒体的定位一致。而20H12识别所有生命周期阶段的119 kDa和122 kDa抗原，在BSF和MCF中抗原定位与鞭毛一致，在PCF中与鞭毛尖端一致，在EMF中与部分细胞质一致。此外，在CLSM和免疫印迹中，20H12均不与布氏冈比亚锥虫、布氏罗得西亚锥虫和伊氏锥虫的抗原发生反应。因此，20H12识别的抗原似乎是刚果锥虫特异性的。尽管对刚果锥虫特异性的119 kDa和122 kDa抗原进行全面表征还需要进一步研究，但mAb 20H12和特异性抗原不仅可能有助于建立刚果锥虫特异性诊断方法，还可能有助于研究寄生虫生命周期中调节分化的分子机制。
