Mineo J R, Khan I A, Kasper L H
Department of Medicine, Dartmouth Medical School, Hanover, New Hampshire 03756.
Exp Parasitol. 1994 Nov;79(3):351-61. doi: 10.1006/expr.1994.1097.
During its intracellular life cycle within the infected host cell, Toxoplasma gondii is able to undergo rapid asexual replication. Neither the mechanism by which the parasite initiates this process nor the requirements for maintaining it are understood. We produced a monoclonal antibody, 1B8, that identifies a parasite antigen of approximate M(r) 97 kDa as determined by SDS-PAGE. The epitope recognized by mAb 1B8 appears as a collection of vesicular structures scattered throughout the cell cytoplasm. When RH strain parasites are incubated with mAb 1B8 in the absence of serum complement, parasite growth is inhibited by > 90% as determined by radioisotope incorporation. Both attachment and invasion assays show that neither of these parasite-host cell interactions are inhibited by the mAb. However, a marked reduction in the number of intracellular rosettes was observed following mAb treatment of the parasites. Viable extracellular parasites are able to endocytose mAb 1B8. Once within the parasite cytosol the antibody recognizes the vesicular structures similar to those observed with fixed parasites. Immunofluorescence assays with Besnoitia jellisoni and Plasmodium falciparum show that the epitope recognized by mAb 1B8 is conserved among Coccidiae but not the kinetoplastid Leishmania.
在被感染的宿主细胞内的细胞内生命周期中,刚地弓形虫能够进行快速无性繁殖。目前尚不清楚寄生虫启动这一过程的机制以及维持该过程的条件。我们制备了一种单克隆抗体1B8,通过SDS-PAGE鉴定出一种分子量约为97 kDa的寄生虫抗原。单克隆抗体1B8识别的表位表现为散布在整个细胞质中的囊泡结构集合。当RH株寄生虫在无血清补体的情况下与单克隆抗体1B8孵育时,通过放射性同位素掺入测定,寄生虫生长受到>90%的抑制。附着和侵袭试验表明,单克隆抗体均不抑制这些寄生虫与宿主细胞的相互作用。然而,在用单克隆抗体处理寄生虫后,观察到细胞内玫瑰花结数量显著减少。有活力的细胞外寄生虫能够内吞单克隆抗体1B8。一旦进入寄生虫胞质溶胶,该抗体就会识别与固定寄生虫中观察到的类似的囊泡结构。用耶氏贝斯诺球虫和恶性疟原虫进行的免疫荧光试验表明,单克隆抗体1B8识别的表位在球虫中保守,但在动基体利什曼原虫中不保守。