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    Immune cells migrating against fluid flow generate significantly higher forces than when static. This study quantifies these forces, revealing a distinct mechanical program for upstream migration and paving the way for future research.

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    Area of Science:

    • Cellular and Molecular Biology
    • Biophysics
    • Immunology

    Background:

    • Cell migration is essential for leukocyte function in tissues.
    • Upstream migration, moving against fluid flow on ICAM-1 surfaces, is mediated by LFA-1 but its mechanical basis is unknown.

    Purpose of the Study:

    • To quantify the mechanical forces generated during upstream migration using Traction Force Microscopy (TFM).
    • To characterize the spatiotemporal force patterns of KG1a cells during upstream migration under shear flow.

    Main Methods:

    • Utilized Traction Force Microscopy (TFM) to measure forces generated by KG1a cells.
    • Applied shear flow to ICAM-1 functionalized hydrogels to induce upstream migration.
    • Analyzed force generation under static and flow conditions.

    Main Results:

    • KG1a cells under shear flow exhibited polarized tractions aligned with migration direction.
    • Maximum RMS traction forces were significantly elevated during upstream migration (428.5 ± 63.0 nN) compared to static conditions (220.8 ± 22.2 nN).
    • Average RMS forces were also higher during upstream migration (82.6 ± 12.9 nN) versus static (45.9 ± 4.4 nN).

    Conclusions:

    • Upstream migration involves amplified stresses to overcome opposing forces, indicating a distinct mechanical program.
    • This study provides the first quantitative force characterization of upstream migration under shear stress.
    • The findings establish a platform for studying molecular regulators of force generation in immune cell trafficking.