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Method of Generating Tolerogenic Maturation-Resistant Dendritic Cells and Testing for Their Immune-Regulatory Functions In Vivo in the Context of Transplantation

  • Sherrie J. Divito
  • Adrian E. MorelliEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1899)

Abstract

During that past two decades, advances in techniques for generating in vitro immune-suppressive dendritic cells (DCs) have heralded the use of these pro-tolerogenic DCs as therapeutics against transplant rejection and autoimmune diseases. In transplantation, previous dogma assumed that systemically administered therapeutic DCs bearing donor antigens (Ags) control the anti-donor response by directly interacting with anti-donor T cells in vivo. However, recent evidence indicates that the exogenously-administered therapeutic DCs instead function as Ag-transporting cells that transfer donor Ags to recipient’s Ag-presenting cells (APCs) for presentation to T cells. In secondary lymphoid organs, presentation of acquired donor Ags by recipient’s quiescent DCs triggers deficient activation and eventual apoptosis of donor-specific effector T cells, leading to a relative increase in the percentage of donor-specific regulatory T cells. This chapter describes the methodology to generate in vitro immune-suppressive DCs that are resistant to maturation, and to assess in vivo both their survival and their ability to regulate donor-specific T cells in a mouse model.

Key words

Tolerogenic dendritic cells Vitamin D3 Cell therapy Transplantation Mouse 

Notes

Acknowledgments

We thank the comments of the Research Specialist William J. Shufesky (Univ. of Pittsburgh, Pittsburgh, PA, USA).

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of DermatologyBrigham and Women’s Hospital and Harvard Medical SchoolBostonUSA
  2. 2.Department of Surgery, T.E. Starzl Transplantation InstituteUniversity of Pittsburgh Medical CenterPittsburghUSA

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