Alderete J F, Demĕs P, Gombosová A, Valent M, Yánoska A, Fabusová H, Kasmala L, Garza G E, Metcalfe E C
Infect Immun. 1987 May;55(5):1037-41. doi: 10.1128/iai.55.5.1037-1041.1987.
Fresh isolates of Trichomonas vaginalis were examined for reactions to a panel of five monoclonal antibodies (MAbs). Four MAbs (C20A3, DM126, DM116, and C55) were to distinct surface immunogens and one MAb (L64) was to a cytoplasmic component. The fresh isolates were evaluated by indirect immunofluorescence (IF), immunoblotting, and radioimmunoprecipitation. IF assay with C20A3 MAb gave isolates which were homogeneous nonstaining (negative [Neg] phenotype) and isolates which were heterogeneous staining and nonstaining (positive [Pos] and Neg phenotype, respectively) organisms. Immunoblotting and radioimmunoprecipitation assays revealed that surface phenotypic heterogeneity among isolates with C20A3 MAb was due to the presence or absence of the immunogen from the parasite surface. IF assay with DM126 MAb also gave Pos and Neg phenotypes among parasites of some isolates. All of the isolates were always Neg phenotype with DM116 and C55 MAbs. The occurrence of Neg phenotype organisms with DM126, DM116, and C55 was due to epitope inaccessibility to their respective MAbs and not to the absence of the immunogen from trichomonal membranes. All isolates possessed the cytoplasmic protein recognized by L64 MAb. Paired isolates (taken 5 to 6 days apart) from 24 women were also studied. Four of the 24 paired isolates (16%) had different phenotype distributions at the two timepoints for C20A3. Fresh isolates also underwent phenotypic variation during in vitro growth and multiplication, as determined with C20A3. Also, 7 of the 24 paired isolates demonstrated dramatic changes in the accessibility of DM126 MAb to epitope binding. Lastly, 55 (90%) of 60 serum samples from patients with trichomoniasis evaluated in this study possessed antibody to the C20A3 reactive molecule. The data show that the fresh T. vaginalis isolates were predominantly Neg phenotype and confirm the occurrence of protein and epitope phenotypic variation for major immunogens among fresh isolates of the pathogenic human trichomonads.
对阴道毛滴虫的新鲜分离株进行了检测,以观察它们对一组五种单克隆抗体(MAb)的反应。四种单克隆抗体(C20A3、DM126、DM116和C55)针对不同的表面免疫原,一种单克隆抗体(L64)针对一种细胞质成分。通过间接免疫荧光(IF)、免疫印迹和放射免疫沉淀对新鲜分离株进行评估。用C20A3单克隆抗体进行的IF检测得到了均一不染色(阴性[Neg]表型)的分离株和异质染色与不染色(分别为阳性[Pos]和Neg表型)的生物体。免疫印迹和放射免疫沉淀检测表明,C20A3单克隆抗体处理的分离株之间的表面表型异质性是由于寄生虫表面免疫原的存在或缺失。用DM126单克隆抗体进行的IF检测在一些分离株的寄生虫中也产生了Pos和Neg表型。所有分离株用DM116和C55单克隆抗体检测时始终为Neg表型。DM126、DM116和C55出现Neg表型生物体是由于其各自的单克隆抗体无法接近表位,而不是由于滴虫膜上缺乏免疫原。所有分离株都具有L64单克隆抗体识别的细胞质蛋白。还研究了来自24名女性的配对分离株(间隔5至6天采集)。24对配对分离株中有4对(16%)在两个时间点对C20A3的表型分布不同。新鲜分离株在体外生长和繁殖过程中也发生了表型变异,这是用C20A3确定的。此外,24对配对分离株中有7对显示DM126单克隆抗体与表位结合的可及性发生了显著变化。最后,本研究中评估的60份滴虫病患者血清样本中有55份(90%)含有针对C20A3反应性分子的抗体。数据表明,新鲜的阴道毛滴虫分离株主要为Neg表型,并证实了致病性人体毛滴虫新鲜分离株中主要免疫原的蛋白质和表位表型变异的发生。