Quantifying spatial dynamics of Mycobacterium tuberculosis infection of human macrophages using microfabricated patterns

Savulescu, AF; Peton, N; Oosthuizen, D; Hazra, R; Rousseau, RP; Mhlanga, MM; Coussens, AK

Author(s): Savulescu, AF; Peton, N; Oosthuizen, D; Hazra, R; Rousseau, RP; Mhlanga, MM; Coussens, AK;
Details: Publication Year 2023-11-07,Volume 3,Issue #11,Page 100640
Journal Title: Cell Reports Methods
Abstract: Macrophages provide a first line of defense against invading pathogens, including the leading cause of bacterial mortality, Mycobacterium tuberculosis (Mtb). A challenge for quantitative characterization of host-pathogen 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 quantitatively 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-γ stimulation increases 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 alterations 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 downstream automated analytical approaches.
Publisher: Elsevier
Keywords: CP: Cell biology; CP: Microbiology; Mdm; Mtoc; Mtb; Nhlrc2; cell wall; host-pathogen interactions; interferon; micropattern; phagocytosis; single cell
Research Division(s): Infectious Diseases And Immune Defence
PubMed ID: 37963461
Publisher's Version: https://doi.org/10.1016/j.crmeth.2023.100640
Open Access at Publisher's Site: https://doi.org/10.1016/j.crmeth.2023.100640
Terms of Use/Rights Notice: Refer to copyright notice on published article.

Creation Date: 2023-11-20 03:32:59

Last Modified: 2023-11-30 09:12:27