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Diabetologia

pp 1–12 | Cite as

Phenotypic alterations in pancreatic lymph node stromal cells from human donors with type 1 diabetes and NOD mice

  • Jorge Postigo-Fernandez
  • Donna L. Farber
  • Rémi J. CreusotEmail author
Article

Abstract

Aims/hypothesis

Tolerance induction in lymph nodes can be mediated by both haematopoietic cells (e.g. specific dendritic cells subsets) and by non-haematopoietic cells (e.g. lymph node stromal cells [LNSCs]) when they present peripheral tissue antigens to autoreactive T cells. LNSCs normally regulate T cell trafficking and survival and help to maintain peripheral tolerance by exerting immunosuppressive effects. However, whether autoimmunity can be associated with defective tolerogenic functions of LNSCs is unknown and studies aimed at characterising LNSCs in humans are lacking. We hypothesised that dysregulated T cell responses in pancreatic lymph nodes (PLNs) from donors with type 1 diabetes and from NOD mice may be associated with altered LNSC function.

Methods

We analysed PLNs from donors with type 1 diabetes and NOD mice for LNSC distribution and phenotype using flow cytometry. We assessed the expression of tolerance-related genes in different subsets of LNSCs from human donors, as well as in a population of dendritic cells enriched in autoimmune regulator (AIRE)+ cells and identified as HLA-DRhigh CD45low.

Results

The relative frequency of different LNSC subsets was altered in both donors with type 1 diabetes and NOD mice, and both MHC class II and programmed death-ligand 1 (PD-L1) expression were upregulated in human type 1 diabetes. Tolerance-related genes showed similar expression profiles between mouse and human LNSCs at steady state but were generally upregulated in the context of human type 1 diabetes, while, at the same time, many such genes were downregulated in the AIRE-enriched dendritic cell population.

Conclusion/interpretation

Our study shows that LNSCs are substantially altered in type 1 diabetes, but, surprisingly, they exhibit an enhanced tolerogenic phenotype along with increased antigen-presenting potential, which may indicate an attempt to offset dendritic cell-related tolerogenic defects in tolerance. Thus, LNSCs could constitute alternative therapeutic targets in which to deliver antigens to help re-establish tolerance and prevent or treat type 1 diabetes.

Data availability

All data generated or analysed during this study are included in the published article (and its online supplementary files). Biomark gene expression data were deposited on the Mendeley repository at https://data.mendeley.com/datasets/d9rdzdmvyf/1. Any other raw datasets are available from the corresponding author on reasonable request. No applicable resources were generated or analysed during the current study.

Keywords

Beta cell antigens Dendritic cells Lymph node stromal cells Pancreatic lymph nodes Tolerance Type 1 diabetes 

Abbreviations

AAb

Autoantibody

AIRE

Autoimmune regulator

APC

Antigen-presenting cell

BEC

Blood (vascular) endothelial cell

DNC

Double-negative cell

FEZF2

FEZ family zinc finger 2

FRC

Fibroblastic reticular cell

LEC

Lymphatic endothelial cell

LNSC

Lymph node stromal cell

MFI

Mean fluorescence intensity

NOR

Non-obese diabetes-resistant

nPOD

Network of Pancreatic Organ Donors with Diabetes

PD-L1

Programmed death-ligand 1

PDPN

Podoplanin

PLN

Pancreatic lymph nodes

TRA

Tissue-restricted antigen

Notes

Acknowledgements

The authors thank A. Pugliese (University of Miami) for critical reading of the manuscript; surgeons N. Matsuoka, D. J. Carpenter and T. Senda (Department of Surgery, Columbia University, New York, NY, USA) who collected tissues procured by LiveOnNY; the nPOD Pathology team; and the organ donors and their families who contributed via nPOD and LiveOnNY. We also thank T. Brusko and H. Seay (University of Florida) for providing residual PLN tissue that helped us to set up and optimise our initial studies.

Contribution statement

JP-F and RJC designed the experiments. JP-F performed the experiments and analysed the data. DLF contributed lymph node samples via LiveOnNY and experimental design insights. RJC directed the research. JP-F and RJC wrote the manuscript. All authors critically edited and approved the manuscript. RJC is the guarantor of this work and, as such, had full access to all the data in the study and takes responsibility for the integrity of the data and the accuracy of the data analysis.

Funding

These studies were funded by a pilot grant from Columbia’s Clinical and Translational Science Award (UL1TR000040), by a JDRF Transition Award (10-2010-790) and by the Helmsley Charitable Trust George S. Eisenbarth nPOD Award for Team Science (2015PG-T1D052). JP-F was supported by a Berrie Fellowship in Diabetes Research from the Berrie Foundation and by a postdoctoral fellowship from the ADA (1-18-PDF-151). The research reported in this publication was supported by nPOD (RRID: SCR_014641), a collaborative type 1 diabetes research project sponsored by the JDRF (nPOD: 5-SRA-2018-557-Q-R) and The Leona M. & Harry B. Helmsley Charitable Trust (Grant no. 2018PG-T1D053). Organ Procurement Organizations partnering with nPOD to provide research resources are listed at www.jdrfnpod.org//for-partners/npod-partners. Studies were also performed using the CCTI Flow Cytometry Core, supported in part by the Office of the Director, National Institutes of Health (awards S10RR027050 and S10OD020056), and by the Diabetes Research Center (grant P30DK063608).

Duality of interest

The authors declare that there is no duality of interest associated with this manuscript.

Supplementary material

125_2019_4984_MOESM1_ESM.pdf (1.8 mb)
ESM 1 (PDF 1878 kb)

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Jorge Postigo-Fernandez
    • 1
    • 2
    • 3
  • Donna L. Farber
    • 1
    • 4
    • 5
  • Rémi J. Creusot
    • 1
    • 2
    • 3
    Email author
  1. 1.Columbia Center for Translational ImmunologyColumbia University Medical CenterNew YorkUSA
  2. 2.Department of MedicineColumbia University Medical CenterNew YorkUSA
  3. 3.Naomi Berrie Diabetes CenterColumbia University Medical CenterNew YorkUSA
  4. 4.Department of SurgeryColumbia University Medical CenterNew YorkUSA
  5. 5.Department of Microbiology & ImmunologyColumbia University Medical CenterNew YorkUSA

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