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Related Experiment Video

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Isolation of Murine Lymph Node Stromal Cells
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Engineered human lymph node stroma model for examining interstitial fluid flow and T cell egress.

Jennifer H Hammel, Abhinav Arneja, Jessica Cunningham

    Biorxiv : the Preprint Server for Biology
    |December 16, 2024
    PubMed
    Summary
    This summary is machine-generated.

    Interstitial fluid flow significantly impacts lymph node (LN) structure and function. This study developed a novel LN model revealing how flow affects stromal cells, T cell migration, and inflammation, crucial for immunoengineering.

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    Area of Science:

    • Immunology
    • Bioengineering
    • Cell Biology

    Background:

    • Lymph nodes (LNs) are vital for immunosurveillance and feature organized stromal cells and immune cells.
    • Fibroblastic reticular cells (FRCs) and lymphatic endothelial cells (LECs) guide T cell migration and are sensitive to fluid flow.
    • Inflammation increases LN interstitial fluid flow, but its impact on cell behavior is poorly understood.

    Purpose of the Study:

    • To engineer a human in vitro model of the lymph node (LN) stroma to investigate the effects of interstitial fluid flow.
    • To examine how varying interstitial flow rates influence LN stromal cell morphology and function.
    • To assess the impact of interstitial flow on T cell migration and inflammatory cytokine secretion within the engineered LN model.

    Main Methods:

    • Developed a 3D engineered human lymph node (LN) stroma model using FRC-laden hydrogels and LEC monolayers.
    • Applied gravity-driven interstitial flow at different rates (0.8 µm/s and 3.0 µm/s) to the engineered LN model.
    • Quantified FRC coverage, LEC barrier integrity, T cell egress, and inflammatory cytokine secretion under varying flow conditions.

    Main Results:

    • High interstitial flow rates enhanced FRC coverage and proliferation but decreased LEC barrier integrity.
    • Interstitial flow, irrespective of magnitude, significantly reduced CD4+ and CD8+ T cell egress from the engineered LN.
    • A flow rate of 3.0 µm/s, but not 0.8 µm/s, correlated with increased inflammatory cytokine secretion.

    Conclusions:

    • Interstitial fluid flow is a critical factor in regulating lymph node (LN) stromal cell morphology and function.
    • Altered interstitial flow dynamics significantly impede T cell migration within the LN microenvironment.
    • This engineered LN model highlights the importance of flow in modulating immune cell behavior and inflammation for improved immunoengineering strategies.