Podoplanin expression in fibroblasts determines lymph node architecture and adaptive immune function

Abstract

Lymph nodes are uniquely organised to form specialised niches for immune interactions. Fibroblastic reticular cells (FRCs) are an essential stromal component of lymph nodes – forming intricate 3-dimensional networks to facilitate communication between immune cells and depositing and ensheathing extracellular matrix on the conduit network. However, beyond these structural roles, FRCs regulate immune function through the production of growth factors, chemokines and inflammatory cues. Here we sought to determine how the immunoregulatory properties of FRCs are determined. Since PDPN has been implicated in lymph node development, we directly tested how the PDPN/CLEC-2 signalling axis impacted the immunoregulatory properties of FRCsin vitroandin vivo. We find that FRCs use the PDPN/CLEC-2 signalling axis to switch transcriptional states and alter the expression of immune related genes.In vivo, genetic deletion of PDPN from fibroblastic stroma in PDGFRαmGFPΔPDPNmice downregulated key immunoregulatory molecules CCL21, VCAM-1 and ICAM-1 and attenuated the activation, proliferation and differentiation of lymphocyte populations. Further, PDGFRαmGFPΔPDPNmice exhibited severe disruption of the FRC network structure, leading to a failure to separate B and T lymphocytes and misdistribution of myeloid cells through the tissue. We conclude that PDPN expression controls signalling pathways beyond cytoskeletal regulation and cell mechanics and that PDPN expression is required for FRC phenotype and function in lymph nodes.