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猫胎儿三毛滴虫分离株中表面抗原TF1.17的评估

Evaluation of surface antigen TF1.17 in feline Tritrichomonas foetus isolates.

作者信息

Gould E N, Corbeil L B, Kania S A, Tolbert M K

机构信息

The University of Tennessee College of Veterinary Medicine, Departments of Small Animal Clinical Sciences and Biomedical and Diagnostic Sciences, Knoxville, TN, United States.

The University of San Diego School of Medicine, San Diego, CA, Department of Pathology, United States.

出版信息

Vet Parasitol. 2017 Sep 15;244:144-153. doi: 10.1016/j.vetpar.2017.08.001. Epub 2017 Aug 3.

Abstract

Tritrichomonas foetus (T. foetus) is a flagellated protozoa that infects the distal ileum and proximal colon of domestic cats, as well as the urogenital tract of cattle. Feline trichomonosis is recognized as a prevalent cause of chronic diarrhea in cats worldwide. The suspected route of transmission is fecal-oral, with cats in densely crowded environments at highest risk for infection. Thus, the recommended strategy for minimizing spread of infection is to identify and isolate T. foetus-positive cats from the general population. Rapid identification of infected cats can be challenging due to the inability to accurately and quickly detect the organism in samples at point of care facilities. Thus, identification of targets for use in development of a novel diagnostic test, as well as a vaccine or therapy for T. foetus infection is a significant area of research. Despite a difference in organ tropism between T. foetus genotypes, evidence exists for conserved virulence factors between feline and bovine T. foetus. The bovine T. foetus surface antigen, TF1.17, is an adhesin that is conserved across isolates. Vaccination with the purified antigen results in amelioration of cytopathogenicity and more rapid clearance of infection in cattle. We previously showed that three feline isolates of T. foetus were positive for TF1.17 antigen so we further hypothesized that TF1.17 is conserved across feline T. foetus isolates and that this antigen would represent an attractive target for development of a novel diagnostic test or therapy for feline trichomonosis. In these studies, we used monoclonal antibodies previously generated against 1.15 and 1.17 epitopes of the bovine T. foetus TF1.17 antigen, to evaluate for the presence and role of TF1.17 in the cytopathogenicity of feline T. foetus. A previously validated in vitro co-culture approach was used to model feline T. foetus infection. Immunoblotting, immunofluorescence assays, and flow cytometric analysis confirmed the presence and surface localization of antigen TF1.17 across all feline T. foetus isolates tested. Antigen TF1.17 was notably absent in the presumably nonpathogenic intestinal trichomonad, Pentatrichomonas hominis, a parasite that can be confused microscopically with T. foetus. Similar to bovine trichomoniasis, TF1.17 was found to promote T. foetus adhesion to the intestinal epithelium. These results support further characterization and development of the TF1.17 antigen as a possible target for the diagnosis and prevention of feline T. foetus infection.

摘要

胎儿三毛滴虫(T. foetus)是一种鞭毛虫原生动物,可感染家猫的回肠末端和结肠近端,以及牛的泌尿生殖道。猫毛滴虫病被认为是全球范围内猫慢性腹泻的常见病因。推测其传播途径为粪口传播,处于密集拥挤环境中的猫感染风险最高。因此,为尽量减少感染传播,推荐的策略是从普通猫群中识别并隔离胎儿三毛滴虫阳性猫。由于无法在医疗点设施中准确快速地检测样本中的病原体,快速识别受感染的猫具有挑战性。因此,确定用于开发新型诊断测试以及胎儿三毛滴虫感染疫苗或疗法的靶点是一个重要的研究领域。尽管胎儿三毛滴虫不同基因型之间存在器官嗜性差异,但有证据表明猫和牛胎儿三毛滴虫之间存在保守的毒力因子。牛胎儿三毛滴虫表面抗原TF1.17是一种在所有分离株中都保守的黏附素。用纯化抗原进行疫苗接种可改善细胞致病性,并使牛体内的感染更快清除。我们之前发现三株猫胎儿三毛滴虫分离株的TF1.17抗原呈阳性,因此我们进一步推测TF1.17在猫胎儿三毛滴虫分离株中是保守的,并且该抗原将成为开发新型猫毛滴虫病诊断测试或疗法的有吸引力的靶点。在这些研究中,我们使用先前针对牛胎儿三毛滴虫TF1.17抗原的1.15和1.17表位产生的单克隆抗体,来评估TF1.17在猫胎儿三毛滴虫细胞致病性中的存在情况和作用。采用先前验证的体外共培养方法来模拟猫胎儿三毛滴虫感染。免疫印迹、免疫荧光分析和流式细胞术分析证实了抗原TF1.17在所测试的所有猫胎儿三毛滴虫分离株中的存在及其表面定位。在可能无致病性的肠道滴虫人五毛滴虫(一种在显微镜下可能与胎儿三毛滴虫混淆的寄生虫)中,明显不存在抗原TF1.17。与牛毛滴虫病相似,发现TF1.17可促进胎儿三毛滴虫黏附于肠上皮。这些结果支持进一步对TF1.17抗原进行表征和开发,将其作为诊断和预防猫胎儿三毛滴虫感染的可能靶点。

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