Percorrer por autor "Spengler, Bernhard"
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- Mass spectrometry imaging of in vitro Cryptosporidium parvum-infected cells and host tissuePublication . Anschütz, Nils H.; Gerbig, Stefanie; Ghezellou, Parviz; Silva, Liliana M. R.; Vélez, Juan Diego; Hermosilla, Carlos R.; Taubert, Anja; Spengler, BernhardCryptosporidium parvum is a zoonotic-relevant parasite belonging to the phylum Alveolata (subphylum Apicomplexa). One of the most zoonotic-relevant etiologies of cryptosporidiosis is the species C. parvum, infecting humans, cattle and wildlife. C. parvum-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 C. parvum-infected host cells as well as parasitized neonatal calf intestines. Sections of C. parvum-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 C. parvum-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 C. parvum-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.
- Mass spectrometry imaging of lipid and metabolite distributions in cysts of Besnoitia besnoiti-Infected bovine skinPublication . Wiedemann, Katja R.; Gerbig, Stefanie; Ghezellou, Parviz; Pilgram, Alejandra; Hermosilla, Carlos; Taubert, Anja; Silva, Liliana M. R.; Spengler, BernhardBovine besnoitiosis is a disease caused by the obligate intracellular parasite Besnoitia besnoiti. During its chronic stage, the parasite forms large, thick-walled cysts of up to 600 μm in diameter in the skin and other tissues. To assess an overview of parasite-induced metabolic changes during chronic infection, B. besnoiti-infected skin samples were analyzed by high-resolution atmospheric-pressure scanning microprobe matrix-assisted laser desorption/ionization mass spectrometry imaging (AP-SMALDI MSI). Overall, infection-driven, significant changes of 467 lipids and metabolites were found in comparison to noninfected control samples. Most of them belong to the group of phosphatidic acids (PAs), phosphatidylserines (PSs), phosphatidylcholines (PCs)/phosphatidylethanolamines (PEs), triacylglycerides (TGs), phosphatidylinositols (PIs) and phosphatidylglycerols (PGs). When these quantitative data were combined with analyses on the lateral distribution of respective infection markers, MS images of significantly changed ion signals with specific lateral distributions were generated, matching with typical biological structures as observed in Hematoxylin and eosin (H&E)-stained tissue sections. Ultrahigh-resolution MALDI MSI with a pixel size of 2 μm and 3-dimensional reconstruction gave further insights into cyst construction.