Project #11642
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#11642 : Defining Shigella-targeting of human lamina propria mononuclear cells using CyTOF technology
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Name of Applicant : Katja Brunner
Date of application : 23-04-2018
Unit : Molecular Microbial Pathogenesis
Location : Duclaux, Bertrand wing, 1ere etage, 06A
Phone : 01 40 61 37 71
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Project context and summary :

Invasion of human intestinal epithelial cells by Shigella flexneri is secondary to the delivery of bacterial effectors into the host cell cytoplasm via a type III secretion system (T3SS). By using a beta-lactamase reporter tool we observed that in contrast to the epithelium, human lymphocytes are mainly targeted by injection of T3SS effectors not resulting in subsequent cell invasion (Pinaud et al., 2017). Furthermore, we observed that the targeting process, in form of successful injection of effectors into the host cell, is dependent on glycan-glycan interactions between bacterial and host cell surfaces rendering the targeting process to be dependent on the activation state of the host cell (Belotserkovsky et al., 2018). CyTOF technology is a research tool used for phenotypic analysis of complex cell population allowing for the simultaneous labelling of up to 40 different surface and intracellular marker without issues of compensation as present in regular flow cytometry (van Unen et al., 2016). Using CyTOF technology and the beta-lactamase reporter tool, we will perform a detailed analysis of Shigella targeting in a complex cell population using human lamina propria mononuclear cells (LPMCs), isolated from human colon explants. Analysis will address the question if specific cellular subsets are preferentially targeted in the intestinal environment and if this differs from targeting of peripheral blood mononuclear cells (PBMCs) diverging in their immune phenotypes and cellular activation.

Related team publications :
Pinaud et al. (2017), PNAS, 114 (37): 9954-9959
Belotserkovsky et al. (2018), mBio, 9(1): e02309-17
van Unen et al. (2016), Immunity, 44: 1-13
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Project Type : Short
Status : Closed
Global Satisfaction for this application : Pretty Good (4/5)

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