Institute of Inorganic and Analytical Chemistry, Justus Liebig University Giessen, 35392 Giessen, Germany.
Institute of Parasitology, Biomedical Research Center Seltersberg, Justus Liebig University Giessen, 35392 Giessen, Germany.
Biomolecules. 2023 Jul 31;13(8):1200. doi: 10.3390/biom13081200.
is a zoonotic-relevant parasite belonging to the phylum Alveolata (subphylum Apicomplexa). One of the most zoonotic-relevant etiologies of cryptosporidiosis is the species , infecting humans, cattle and wildlife. -infected intestinal mucosa as well as host cells infected in vitro have not yet been the subject of extensive biochemical investigation. Efficient treatment options or vaccines against cryptosporidiosis are currently not available. Human cryptosporidiosis is currently known as a neglected poverty-related disease (PRD), being potentially fatal in young children or immunocompromised patients. In this study, we used a combination of atmospheric pressure scanning microprobe matrix-assisted laser desorption/ionization (AP-SMALDI) mass spectrometry imaging (MSI) and liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) to determine and locate molecular biomarkers in in vitro -infected host cells as well as parasitized neonatal calf intestines. Sections of -infected and non-infected host cell pellets and infected intestines were examined to determine potential biomarkers. Human ileocecal adenocarcinoma cells (HCT-8) were used as a suitable in vitro host cell system. More than a thousand different molecular signals were found in both positive- and negative-ion mode, which were significantly increased in -infected material. A database search in combination with HPLC-MS/MS experiments was employed for the structural verification of markers. Our results demonstrate some overlap between the identified markers and data obtained from earlier studies on other apicomplexan parasites. Statistically relevant biomarkers were imaged in cell layers of -infected and non-infected host cells with 5 µm pixel size and in bovine intestinal tissue with 10 µm pixel size. This allowed us to substantiate their relevance once again. Taken together, the present approach delivers novel metabolic insights on neglected cryptosporidiosis affecting mainly children in developing countries.
是一种属亲动物寄生虫，属于纤毛门（子孢子亚门）。隐孢子虫病最相关的人畜共患病原之一是 ，感染人类、牛和野生动物。-感染的肠黏膜以及体外感染的宿主细胞尚未成为广泛生化研究的对象。目前尚无有效的隐孢子虫病治疗方法或疫苗。人类隐孢子虫病目前被认为是一种被忽视的与贫困相关的疾病（PRD），在幼儿或免疫功能低下的患者中可能致命。在这项研究中，我们使用大气压扫描微探针基质辅助激光解吸/电离（AP-SMALDI）质谱成像（MSI）和液相色谱-串联质谱（HPLC-MS/MS）的组合来确定和定位体外感染的宿主细胞以及寄生的新生牛肠中的分子生物标志物。检查感染和未感染的宿主细胞颗粒和感染的肠段以确定潜在的生物标志物。我们使用人回肠腺癌细胞（HCT-8）作为合适的体外宿主细胞系统。在正离子和负离子模式下都发现了超过一千种不同的分子信号，这些信号在感染材料中显著增加。结合 HPLC-MS/MS 实验的数据库搜索用于对标记物进行结构验证。我们的结果表明，鉴定出的标记物与其他顶复门寄生虫的早期研究数据之间存在一些重叠。以 5 µm 像素大小对感染和未感染宿主细胞的细胞层以及以 10 µm 像素大小对牛肠组织中的统计上相关的生物标志物进行成像。这再次证实了它们的相关性。总的来说，这种方法提供了关于主要影响发展中国家儿童的被忽视的隐孢子虫病的新的代谢见解。
