Poster Presentation Lorne Infection and Immunity 2023

Establishing human small intestinal monolayers to study EPEC infection (#113)

Eva Chan 1 2 , Cristina Giogha 1 2 , Paul Hertzog 1 2 , Elizabeth Hartland 1 2
  1. Centre for Innate Immunity and Infectious Diseases , Hudson Institute of Medical Research, 27-31 Wright Street, Clayton , VIC 3168, Australia
  2. Department of Molecular and Translational Science, Monash University, Clayton, Victoria, Australia

The gastrointestinal tract is lined by a single layer of epithelial cells that forms a barrier to the external environment. With regular exposure to the external environment, the epithelial barrier is exposed to a number of stressors. An example of this is the introduction of gastrointestinal pathogens, such as enteropathogenic Escherichia coli (EPEC). This bacterium can disrupt the intestinal epithelial lining leading to symptoms such as diarrhoea, fever, vomiting, and in extreme cases death. This is possible due to the bacterial effector proteins that are injected into the host cell resulting in the remodelling of host cell cytoskeleton and changes to host cell signalling pathways.

Such findings have been possible due to the use of immortalised cell lines in the laboratory. However, immortalised cell lines do not recapitulate the epithelial cell heterogeneity that is exhibited in the human gastrointestinal tract. Therefore, to better understand the effect of EPEC infection on human intestinal epithelial cells, the organoid culturing system will be utilised. A protocol has been established to grow these cells as a monolayer to allow access to the apical cell surface by EPEC, where immunofluorescent staining and imaging has been used to visualise the interaction. By establishing this model, this will provide further insight to host cell response and epithelial cell function following infection.