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Abstract: TH-OR48

Using Three-Dimensional Imaging and Single-Cell Transcriptomics to Interrogate Human Kidney Lymphatics in Transplant Rejection

Session Information

Category: Transplantation

  • 2001 Transplantation: Basic

Authors

  • Jafree, Daniyal J., UCL Great Ormond Street Institute of Child Health, London, United Kingdom
  • Stewart, Benjamin James, University of Cambridge, Cambridge, Cambridgeshire, United Kingdom
  • Joannou, Maria K., UCL Great Ormond Street Institute of Child Health, London, United Kingdom
  • Russell, Lauren G., UCL Great Ormond Street Institute of Child Health, London, United Kingdom
  • Pomeranz, Gideon, UCL Great Ormond Street Institute of Child Health, London, United Kingdom
  • Scambler, Peter, UCL Great Ormond Street Institute of Child Health, London, United Kingdom
  • Clatworthy, Menna R., University of Cambridge, Cambridge, Cambridgeshire, United Kingdom
  • Motallebzadeh, Reza, UCL Division of Surgery and Interventional Science, London, United Kingdom
  • Long, David A., UCL Great Ormond Street Institute of Child Health, London, United Kingdom

Group or Team Name

  • UCL ICH Kidney Development & Disease Group
Background

Lymphatics participate in immune homeostasis and their dysfunction has been linked to autoimmunity and cancer. There is a need to enhance our understanding of the spatial and molecular features of lymphatics in human kidney health and transplant rejection.

Methods

Wholemount immunolabelling, tissue clearing and 3D microscopy were used to visualise lymphatics in non-transplanted donor kidney and allografts with chronic transplant rejection (CKTR). Furthermore, we integrated multiple human kidney single-cell RNA sequencing (scRNA-seq) datasets, including samples with distinct aetiologies of CKTR.

Results

In donor kidneys, lymphatics reside hierarchically within the cortex, form terminal branches along cortical nephron segments and possess a unique capillary phenotype, which is distinct from other organ lymphatics due to their low expression of LYVE1. In CKTR, lymphatics undergo expansion, lose structural hierarchy and infiltrate the medulla (Fig.1). Allograft lymphatics are predominantly donor-derived, express HLA-DR, and exhibit C4d immunoreactivity; indicative of targetting by anti-allograft antibodies. Additionally, kidney lymphatics are T cell-rich conduits which interconnect tertiary lymphoid structures. Using scRNA-seq, we identify putative crosstalk between lymphatics and T cells, featuring co-inhibitory immune checkpoints.

Conclusion

Utilising 3D imaging and scRNA-seq, we uncovered the spatial and molecular profile of human kidney lymphatics. We have revealed fundamentals of these vessels to inform future studies into renal biology, as well as identifying lymphatic phenomena and molecular candidates involved in CKTR.

Fig.1. The lymphatic phenotype in transplant rejection