Mass Spectrometry Imaging of In Vitro Cryptosporidium parvum-Infected Cells and Host Tissue
Nils H Anschütz1, Stefanie Gerbig1, Parviz Ghezellou1
1Institute of Inorganic and Analytical Chemistry, Justus Liebig University Giessen, 35392 Giessen, Germany.
Abstract:
Cryptosporidium 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.
Insights
Cryptosporidium parvum causes cryptosporidiosis, a neglected disease. This study identified novel molecular biomarkers in infected cells and intestines using mass spectrometry imaging, offering new insights for treatment.
Area of Science:
- Parasitology
- Biochemistry
- Mass Spectrometry Imaging
Background:
- Cryptosporidium parvum is a zoonotic parasite causing cryptosporidiosis, a neglected poverty-related disease.
- Current treatment and vaccine options for cryptosporidiosis are limited.
- Biochemical investigations of C. parvum-infected host cells and intestinal mucosa are scarce.
Purpose of the Study:
- To identify and locate molecular biomarkers in C. parvum-infected host cells and bovine intestinal tissue.
- To gain novel metabolic insights into cryptosporidiosis.
Main Methods:
- Utilized atmospheric pressure scanning microprobe matrix-assisted laser desorption/ionization (AP-SMALDI) mass spectrometry imaging (MSI).
- Employed liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) for structural verification of biomarkers.
- Analyzed in vitro infected human ileocecal adenocarcinoma cells (HCT-8) and neonatal calf intestines.
Main Results:
- Identified over a thousand molecular signals, significantly increased in C. parvum-infected samples.
- Confirmed overlap with previously identified markers in other apicomplexan parasites.
- Successfully imaged statistically relevant biomarkers in host cells (5 µm pixel size) and bovine intestinal tissue (10 µm pixel size).
Conclusions:
- The study provides novel metabolic insights into C. parvum infection.
- Identified molecular biomarkers hold potential for future diagnostic and therapeutic strategies.
- Highlights the need for further research into neglected tropical diseases like cryptosporidiosis.
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