Image analysis of the adhesion of the probiotic strain E. coli Nissle to the fungal pathogen C. albicans using JIPipe

www.leibniz-hki.de References Image analysis of the adhesion of the probiotic strain E. coli Nissle to the fungal pathogen C. albicans using JIPipe Carl-Magnus Svensson1, Alexandre Chamas2,3, Carla Maneira2,4,5, Marc Thilo Figge1,3,4, Gerald Lackner2,3,5 1Applied Systems Biology, Leibniz Institute for Natural Product Research and Infection Biology –Hans Knöll Institute, Jena, Germany 2Junior Research Group Synthetic Microbiology, Leibniz-Institute for Natural Product Research and Infection Biology, Jena, Germany 3Cluster of Excellence Balance of the Microverse, Friedrich Schiller University Jena, Jena, Germany 4 Institute of Microbiology, Faculty of Biological Sciences, Friedrich-Schiller University Jena, Jena, Germany 5Chair of Biochemistry of Microorganisms, Faculty of Life Sciences: Food Nutrition and Health, University of Bayreuth, Bayreuth, Germany [1] Chamas, A. et al. Engineering Adhesion of the Probiotic Strain Escherichia coli Nissle to the Fungal Pathogen Candida albicans. ACS Synth. Biol. (2024) [2] Gerst, R., Cseresnyés, Z. & Figge, M.T. JIPipe: visual batch processing for ImageJ. Nat. Methods (2023) Probiotic E. coli Nissle 1917 and Candida albicans Imaging and analysis Conclusions •Fungal infections are responsible for the death of 1.6 million people each year •Live biotherapeutic products (LBPs) as an alternative to conventional antibiotics-based therapy •E. coli Nissle (EcN) targeted towards one of the most prevalent causes of fungal infection, C. albicans •Engineer the display of membrane proteins with affinity for the hyphal form of C. albicans •Six conditions: Control, Actinohivin (EcN/act), Chitinase 92 (EcN/chi), Dectin-2 (EcN/dec), Synthetic peptide 01 (EcN/SP1) and Synthetic peptide 02 (EcN/SP2) [1] •Synthetic peptide SP1 has the same sequence length as the chitinase 92 binding domain and many amino acids •Carbohydrate binding partially relies on interactions involving aromatic amino acids •SP2 possesses the same sequence as SP1 but with all aromatic amino acids exchanged for non-aromatic ones carl-magnus.sv[email protected] Funded by the German Federal Ministry of Education and Research within the funding program Photonics Research Germany, Project Leibniz Center for Photonics in Infection Research, Subproject LPI-BT3, contract number 13N15709 •Imaging on an Axiocam 202 (Carl Zeiss Microscopy Deutschland GmbH, Oberkochen, Germany) camera (0.83x magnification, final magnification 33.2x) •C. albicans hyphae were imaged using brightfield mode and E. coli Nissle was labelled by DYKDDDDK antibody (red) •Custom pipeline developed in JIPipe [2] to segment hyphae and bacteria •JIPipe is a powerful tool with pipelines that are easy to decipher and adapt •Western blot analysis of protein content •EcN/chi, EcN/SP1 and EcN/SP2 have significantly increased binding proteins •Data analysis directly in JIPipe •EcN/SP2 do not show increased hyphae binding despite an increase in linkers