Interstitial fluid flow in an engineered human lymph node stroma model modulates T cell egress and stromal change

The lymph node (LN) performs essential roles in immunosurveillance throughout the body. Developing in vitro models of this key tissue is of great importance to enhancing physiological relevance in immunoengineering. The LN consists of stromal populations and immune cells, which are highly organized...

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Main Authors: Jennifer H. Hammel, Abhinav Arneja, Jessica Cunningham, Maosen Wang, Sophia Schumaecker, Yamilet Macias Orihuela, Tochukwu Ozulumba, Jonathan M. Zatorski, Thomas J. Braciale, Chance John Luckey, Rebecca R. Pompano, Jennifer M. Munson
Format: Article
Language:English
Published: AIP Publishing LLC 2025-06-01
Series:APL Bioengineering
Online Access:http://dx.doi.org/10.1063/5.0247363
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author Jennifer H. Hammel
Abhinav Arneja
Jessica Cunningham
Maosen Wang
Sophia Schumaecker
Yamilet Macias Orihuela
Tochukwu Ozulumba
Jonathan M. Zatorski
Thomas J. Braciale
Chance John Luckey
Rebecca R. Pompano
Jennifer M. Munson
author_facet Jennifer H. Hammel
Abhinav Arneja
Jessica Cunningham
Maosen Wang
Sophia Schumaecker
Yamilet Macias Orihuela
Tochukwu Ozulumba
Jonathan M. Zatorski
Thomas J. Braciale
Chance John Luckey
Rebecca R. Pompano
Jennifer M. Munson
author_sort Jennifer H. Hammel
collection DOAJ
description The lymph node (LN) performs essential roles in immunosurveillance throughout the body. Developing in vitro models of this key tissue is of great importance to enhancing physiological relevance in immunoengineering. The LN consists of stromal populations and immune cells, which are highly organized and bathed in constant interstitial fluid flow (IFF). The stroma, notably the fibroblastic reticular cells (FRCs) and the lymphatic endothelial cells (LECs), play crucial roles in guiding T cell migration and are known to be sensitive to fluid flow. During inflammation, interstitial fluid flow rates drastically increase in the LN. It is unknown how these altered flow rates impact crosstalk and cell behavior in the LN, and most existing in vitro models focus on the interactions between T cells, B cells, and dendritic cells rather than with the stroma. To address this gap, we developed a human engineered model of the LN stroma consisting of FRC-laden hydrogel above a monolayer of LECs in a tissue culture insert with gravity-driven interstitial flow. We found that FRCs had enhanced coverage and proliferation in response to high flow rates, while LECs experienced decreased barrier integrity. We added CD4+ and CD8+ T cells and found that their egress was significantly decreased in the presence of interstitial flow, regardless of magnitude. Interestingly, 3.0 μm/s flow, but not 0.8 μm/s flow, correlated with enhanced inflammatory cytokine secretion in the LN stroma. Overall, we demonstrate that interstitial flow is an essential consideration in the lymph node for modulating LN stroma morphology, T cell migration, and inflammation.
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spelling doaj-art-ab9150ed7bba4013a1dccd70f7f1bcb12025-08-20T03:28:52ZengAIP Publishing LLCAPL Bioengineering2473-28772025-06-0192026105026105-1410.1063/5.0247363Interstitial fluid flow in an engineered human lymph node stroma model modulates T cell egress and stromal changeJennifer H. Hammel0Abhinav Arneja1Jessica Cunningham2Maosen Wang3Sophia Schumaecker4Yamilet Macias Orihuela5Tochukwu Ozulumba6Jonathan M. Zatorski7Thomas J. Braciale8Chance John Luckey9Rebecca R. Pompano10Jennifer M. Munson11Virginia Tech Fralin Biomedical Research Institute, 4 Riverside Circle, Roanoke, Virginia 24016, USAUniversity of Virginia Department of Pathology, 415 Lane Road, Charlottesville, Virginia 22908, USAVirginia Tech Fralin Biomedical Research Institute, 4 Riverside Circle, Roanoke, Virginia 24016, USAVirginia Tech Fralin Biomedical Research Institute, 4 Riverside Circle, Roanoke, Virginia 24016, USAVirginia Tech Fralin Biomedical Research Institute, 4 Riverside Circle, Roanoke, Virginia 24016, USAVirginia Tech Fralin Biomedical Research Institute, 4 Riverside Circle, Roanoke, Virginia 24016, USAUniversity of Virginia Department of Chemistry, 409 McCormick Road, Charlottesville, Virginia 22904, USAUniversity of Virginia Department of Chemistry, 409 McCormick Road, Charlottesville, Virginia 22904, USAUniversity of Virginia Department of Pathology, 415 Lane Road, Charlottesville, Virginia 22908, USAUniversity of Virginia Department of Pathology, 415 Lane Road, Charlottesville, Virginia 22908, USAUniversity of Virginia Department of Chemistry, 409 McCormick Road, Charlottesville, Virginia 22904, USAVirginia Tech Fralin Biomedical Research Institute, 4 Riverside Circle, Roanoke, Virginia 24016, USAThe lymph node (LN) performs essential roles in immunosurveillance throughout the body. Developing in vitro models of this key tissue is of great importance to enhancing physiological relevance in immunoengineering. The LN consists of stromal populations and immune cells, which are highly organized and bathed in constant interstitial fluid flow (IFF). The stroma, notably the fibroblastic reticular cells (FRCs) and the lymphatic endothelial cells (LECs), play crucial roles in guiding T cell migration and are known to be sensitive to fluid flow. During inflammation, interstitial fluid flow rates drastically increase in the LN. It is unknown how these altered flow rates impact crosstalk and cell behavior in the LN, and most existing in vitro models focus on the interactions between T cells, B cells, and dendritic cells rather than with the stroma. To address this gap, we developed a human engineered model of the LN stroma consisting of FRC-laden hydrogel above a monolayer of LECs in a tissue culture insert with gravity-driven interstitial flow. We found that FRCs had enhanced coverage and proliferation in response to high flow rates, while LECs experienced decreased barrier integrity. We added CD4+ and CD8+ T cells and found that their egress was significantly decreased in the presence of interstitial flow, regardless of magnitude. Interestingly, 3.0 μm/s flow, but not 0.8 μm/s flow, correlated with enhanced inflammatory cytokine secretion in the LN stroma. Overall, we demonstrate that interstitial flow is an essential consideration in the lymph node for modulating LN stroma morphology, T cell migration, and inflammation.http://dx.doi.org/10.1063/5.0247363
spellingShingle Jennifer H. Hammel
Abhinav Arneja
Jessica Cunningham
Maosen Wang
Sophia Schumaecker
Yamilet Macias Orihuela
Tochukwu Ozulumba
Jonathan M. Zatorski
Thomas J. Braciale
Chance John Luckey
Rebecca R. Pompano
Jennifer M. Munson
Interstitial fluid flow in an engineered human lymph node stroma model modulates T cell egress and stromal change
APL Bioengineering
title Interstitial fluid flow in an engineered human lymph node stroma model modulates T cell egress and stromal change
title_full Interstitial fluid flow in an engineered human lymph node stroma model modulates T cell egress and stromal change
title_fullStr Interstitial fluid flow in an engineered human lymph node stroma model modulates T cell egress and stromal change
title_full_unstemmed Interstitial fluid flow in an engineered human lymph node stroma model modulates T cell egress and stromal change
title_short Interstitial fluid flow in an engineered human lymph node stroma model modulates T cell egress and stromal change
title_sort interstitial fluid flow in an engineered human lymph node stroma model modulates t cell egress and stromal change
url http://dx.doi.org/10.1063/5.0247363
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