Mass Spectrometry Imaging of In Vitro Cryptosporidium parvum-Infected Cells and Host Tissue

Nils H Anschütz; Stefanie Gerbig; Parviz Ghezellou; Liliana M R Silva; Juan Diego Vélez; Carlos R Hermosilla; Anja Taubert; Bernhard Spengler

doi:10.3390/biom13081200

Mass Spectrometry Imaging of In Vitro Cryptosporidium parvum-Infected Cells and Host Tissue

Biomolecules. 2023 Jul 31;13(8):1200. doi: 10.3390/biom13081200.

Authors

Nils H Anschütz¹, Stefanie Gerbig¹, Parviz Ghezellou¹, Liliana M R Silva^{2

3

4}, Juan Diego Vélez², Carlos R Hermosilla², Anja Taubert², Bernhard Spengler¹

Affiliations

¹ Institute of Inorganic and Analytical Chemistry, Justus Liebig University Giessen, 35392 Giessen, Germany.
² Institute of Parasitology, Biomedical Research Center Seltersberg, Justus Liebig University Giessen, 35392 Giessen, Germany.
³ Egas Moniz Center for Interdisciplinary Research (CiiEM), Egas Moniz School of Health & Science, 2829-511 Caparica, Portugal.
⁴ MED-Mediterranean Institute for Agriculture, Environment and Development & CHANGE-Global Change and Sustainability Institute, Institute for Advanced Studies and Research, Universidade de Évora, 7006-554 Évora, Portugal.

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.

Keywords: AP-SMALDI; Cryptosporidium parvum; mass spectrometry imaging.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cattle
Child
Child, Preschool
Cryptosporidiosis*
Cryptosporidium parvum*
Cryptosporidium*
Diagnostic Imaging
Humans
Tandem Mass Spectrometry

Grants and funding

Financial support from the Federal State of Hesse, LOEWE Center DRUID (Novel Drug Targets against Poverty-Related and Neglected Tropical Diseases), the German ‘Bundesministerium für Bildung und Forschung’ (BMBF; DLR 01DG20023; CryptoNETs) and the Deutsche Forschungsgemeinschaft (DFG; Sp314/23-1) is gratefully acknowledged. We would like to thank the Hans-Böckler-Stiftung for granting Nils Anschütz a scholarship. Instrumental support from TransMIT GmbH and Thermo Fisher Scientific is gratefully acknowledged.