An Organotypic Human Lymph Node Model Reveals the Importance of Fibroblastic Reticular Cells for Dendritic Cell Function

Andrew I. Morrison, Aleksandra M. Mikula, Sander W. Spiekstra, Michael de Kok, Alsya J. Affandi, Henk P. Roest, Luc J.W. van der Laan, Charlotte M. de Winde, Jasper J. Koning, Susan Gibbs, Reina E. Mebius

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    3 Citations (Scopus)

    Abstract

    Background: Human lymph node (HuLN) models have emerged with invaluable potential for immunological research and therapeutic application given their fundamental role in human health and disease. While fibroblastic reticular cells (FRCs) are instrumental to HuLN functioning, their inclusion and recognition of importance for organotypic in vitro lymphoid models remain limited. Methods: Here, we established an in vitro three-dimensional (3D) model in a collagen-fibrin hydrogel with primary FRCs and a dendritic cell (DC) cell line (MUTZ-3 DC). To study and characterise the cellular interactions seen in this 3D FRC-DC organotypic model compared to the native HuLN; flow cytometry, immunohistochemistry, immunofluorescence and cytokine/chemokine analysis were performed. Results: FRCs were pivotal for survival, proliferation and localisation of MUTZ-3 DCs. Additionally, we found that CD1a expression was absent on MUTZ-3 DCs that developed in the presence of FRCs during cytokine-induced MUTZ-3 DC differentiation, which was also seen with primary monocyte-derived DCs (moDCs). This phenotype resembled HuLN-resident DCs, which we detected in primary HuLNs, and these CD1a MUTZ-3 DCs induced T cell proliferation within a mixed leukocyte reaction (MLR), indicating a functional DC status. FRCs expressed podoplanin (PDPN), CD90 (Thy-1), CD146 (MCAM) and Gremlin-1, thereby resembling the DC supporting stromal cell subset identified in HuLNs. Conclusion: This 3D FRC-DC organotypic model highlights the influence and importance of FRCs for DC functioning in a more realistic HuLN microenvironment. As such, this work provides a starting point for the development of an in vitro HuLN.

    Original languageEnglish
    Pages (from-to)455-471
    Number of pages17
    JournalTissue Engineering and Regenerative Medicine
    Volume21
    Issue number3
    Early online date19 Dec 2023
    DOIs
    Publication statusPublished - Apr 2024

    Keywords

    • Dendritic cells
    • Fibroblastic reticular cells
    • Human immunology
    • Lymph nodes
    • Organotypic models

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