Human Immune Response Triggered by Entamoeba histolytica in a 3D-Intestinal Model
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Entamoeba histolytica, the agent of amoebiasis, colonizes the human colon and can invade the lining of the colon to disseminate in the deep layers of the intestine. Amoebiasis mainly affects poor people in developing countries, where the barriers between human feces and food or water are inadequate. Humans are the only reservoir of E. histolytica and are the sole target organism of the development of the disease, which limits our knowledge of the crosstalk between the colon and the parasite, especially during the acute phase of infection. In the present work, we constructed an in vitro model of intestinal epithelium that includes an immune component to mimic the immune response against pathogenic microorganisms such as E. histolytica. Using this model and leading-edge technologies, including tissue and cell imaging, transcriptomics, proteomics and ELISA, we investigated the early stages of amoebic infection, in particular, the early immune response. The data obtained highlight the importance of several previously showed virulence markers in patients and experimental models. In addition, we underscored the involvement of other factors that appear to be key regulators of gene expression in the cellular stress responses against amoebiasis and we found novel regulatory mechanisms used by this parasite to modulate the immune response and survive within the human intestine.
KeywordsEntamoeba histolytica 3D-intestinal model Cytokines Mucin Transcriptomics Proteomics
Tumor Necrosis Factor alpha
Phorbol 12-myristate 13-acetate
Phorbol 12-myristate 13-acetate
Amoebic cysteine protease A5
Macrophage-colony stimulating factor
Enzyme-linked immunosorbent assay
Pathogen-associated molecular patterns
- LC-MS/MS proteomics
Liquid chromatography coupled to mass spectrometry
Surface amoebic protein
Macrophage migration inhibitory factor
Second Harmonic Generation signal
The authors gratefully acknowledge Professor Jost Enninga from the Dynamics of Host Pathogen Interactions Unit, Institut Pasteur, Paris, France, for his kind supply of the cell lines, Caco-2/TC7 and HT29-MTX. Special thanks are due to Maria Manich, BIA Unit-Institut Pasteur, for her support in laboratory organization. The authors express their gratitude to all members of Icy software group from BIA Unit-Institut Pasteur, Paris, France, for their help in image analysis. We acknowledge the generous support of “DIM Ile de France” for the support in the acquisition of the two-photon microscope.
The project has received funding from the European ERA-NET Infect-ERA program AMOEBAC (French National Agency for Research (ANR) grants ANR-14-IFEC-0001-01 and ANR-14-IFEC-0001-02). Biomics Platform (Institute Pasteur) is supported by France Génomique (ANR-10-INBS-09-09) and IBISA. AAR is a recipient of Research Career Development Awards from Fundación-IMSS, México. SCC received a postdoctoral fellowship with number 173697 from the program S190-Conacyt, 2017, Mexico.
Conflict of Interest
All the authors declare no potential conflicts of interest.
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