Juliett Anders (Hamburg / DE), Antonia Müller (Berlin / DE), Dr. David Holthaus (Berlin / DE), Dr. Constantin König (Hamburg / DE), Dr. Christian Klotz (Berlin / DE), Prof. Dr. Iris Bruchhaus (Hamburg / DE)
Abstract text
Juliett Anders1, Antonia Müller2, David Holthaus2, Constantin König1, Christian Klotz2, Iris Bruchhaus1
1Bernhard Nocht Institute for Tropical Medicine, Host Parasite Interaction, Hamburg, Germany; 2Robert Koch Institute, Mycotic and Parasitic Agents and Mycobacteria, Berlin, Germany
Introduction
Entamoeba histolytica is a protozoan parasite which causes the disease amoebiasis in humans. In 90 % of infections, the parasite persists in the intestine without causing symptoms, with the amoebae becoming invasive in 10 % of cases. E. histolytica can reach other organs, mainly the liver, through the bloodstream and form abscesses there (1 % of cases). If left untreated, theses abscesses can lead to the death of the host. The reasons why E. histolytica is sometimes invasive and sometimes non-invasive are not yet fully understood. To get a better insight into the processes behind the tissue invasion of E. histolytica, we are using an intestinal organoid derived 2D monolayer model.
Materials & methods
Organoids of the small and the large intestine were grown from intestinal crypts containing adult stem cells and maintained in culture. Organoid derived single cells were seeded onto a filter membrane in a transwell system, where they formed a monolayer after several days. The organoid derived monolayers were characterized via immunofluorescence assays (IFA) and via measurement of the transepithelial electrical resistance (TEER).
Infection of the organoid derived monolayers with the non-pathogenic A1 amoeba clone and with the pathogenic B2 amoeba clone was performed. The infection was analyzed using TEER measurement, IFAs and dual RNAseq.
Results
The data show a decrease in the TEER value within several hours of infection for both amoeba clones.