| Autor: |
Savulescu, Anca F., Peton, Nashied, Oosthuizen, Delia, Hazra, Rudranil, Rousseau, Robert P., Mhlanga, Musa M., Coussens, Anna K. |
| Zdroj: |
Cell Reports Methods; November 2023, Vol. 3 Issue: 11 |
| Abstrakt: |
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 NHLRC2or bacterial surface lipid removal. We identify host organelle position alterations within infected MDMs and differences in Mtbsubcellular 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. |
| Databáze: |
Supplemental Index |
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