@article{Savulescu2023,
abstract = {Macrophages provide a first line of defense against invading pathogens, including the leading cause of bac- terial mortality, Mycobacterium tuberculosis (Mtb). A challenge for quantitative characterization of host-path- ogen processes in differentially polarized primary human monocyte-derived macrophages (MDMs) is their heterogeneous morphology. Here, we describe the use of microfabricated patterns that constrain the size and shape of cells, mimicking the physiological spatial confinement cells experience in tissues, to quantita- tively characterize interactions during and after phagocytosis at the single-cell level at high resolution. Comparing pro-inflammatory (M1) and anti-inflammatory (M2) MDMs, we find interferon-g stimulation in- creases the phagocytic contraction, while contraction and bacterial uptake decrease following silencing of phagocytosis regulator NHLRC2 or bacterial surface lipid removal. We identify host organelle position alter- ations within infected MDMs and differences in Mtb subcellular localization in line with M1 and M2 cellular polarity. Our approach can be adapted to study other host-pathogen interactions and coupled with down- stream automated analytical approaches.},
author = {Savulescu, Anca F and Peton, Nashied and Oosthuizen, Delia and Hazra, Rudranil and Rousseau, Robert P and Mhlanga, Musa M and Coussens, Anna K},
doi = {10.1016/J.CRMETH.2023.100640},
file = {:C$\backslash$:/Users/01462563/AppData/Local/Mendeley Ltd./Mendeley Desktop/Downloaded/Savulescu et al. - 2023 - Quantifying spatial dynamics of iMycobacterium tuberculosisi infection of human macrophages using microfabrica.pdf:pdf},
issn = {2667-2375},
journal = {Cell Reports Methods},
keywords = {OA,fund{\_}ack,original},
mendeley-tags = {OA,fund{\_}ack,original},
month = {nov},
number = {11},
pages = {100640},
pmid = {37963461},
publisher = {Cell Press},
title = {{Quantifying spatial dynamics of \textit{Mycobacterium tuberculosis} infection of human macrophages using microfabricated patterns}},
volume = {3},
year = {2023}
}

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Here, we describe the use of microfabricated patterns that constrain the size and shape of cells, mimicking the physiological spatial confinement cells experience in tissues, to quantita- tively characterize interactions during and after phagocytosis at the single-cell level at high resolution. Comparing pro-inflammatory (M1) and anti-inflammatory (M2) MDMs, we find interferon-g stimulation in- creases the phagocytic contraction, while contraction and bacterial uptake decrease following silencing of phagocytosis regulator NHLRC2 or bacterial surface lipid removal. We identify host organelle position alter- ations within infected MDMs and differences in Mtb subcellular localization in line with M1 and M2 cellular polarity. 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