Go to The Journal of Clinical Investigation
  • About
  • Editors
  • Consulting Editors
  • For authors
  • Publication ethics
  • Transfers
  • Advertising
  • Job board
  • Contact
  • Current issue
  • Past issues
  • By specialty
    • COVID-19
    • Cardiology
    • Immunology
    • Metabolism
    • Nephrology
    • Oncology
    • Pulmonology
    • All ...
  • Videos
  • Collections
    • Resource and Technical Advances
    • Clinical Medicine
    • Reviews
    • Editorials
    • Perspectives
    • Top read articles
  • JCI This Month
    • Current issue
    • Past issues

  • Current issue
  • Past issues
  • Specialties
  • In-Press Preview
  • Editorials
  • Viewpoint
  • Top read articles
  • About
  • Editors
  • Consulting Editors
  • For authors
  • Publication ethics
  • Transfers
  • Advertising
  • Job board
  • Contact
Chimeric antigen receptor costimulation domains modulate human regulatory T cell function
Angela C. Boroughs, … , Shadmehr Demehri, Marcela V. Maus
Angela C. Boroughs, … , Shadmehr Demehri, Marcela V. Maus
Published March 14, 2019
Citation Information: JCI Insight. 2019;4(8):e126194. https://doi.org/10.1172/jci.insight.126194.
View: Text | PDF
Research Article Immunology

Chimeric antigen receptor costimulation domains modulate human regulatory T cell function

  • Text
  • PDF
Abstract

Tregs are key modulators of inflammation and are important for the maintenance of peripheral tolerance. Adoptive immunotherapy with polyclonal Tregs holds promise in organ transplantation, graft-versus-host disease, and autoimmune diseases but may be enhanced by antigen-specific, long-lived Tregs. We modified primary human Tregs with chimeric antigen receptors (CARs) bearing different costimulatory domains and performed in vitro analyses of their phenotype and function. While neither the presence of a CAR nor the type of costimulation domain influenced Foxp3 expression in Tregs, the costimulation domain of the CARs affected CAR-Treg surface phenotype and functions, such as cytokine production. Furthermore, signaling from the CD28 costimulation domain maintained CAR-Treg suppressor function, whereas 4-1B costimulation did not. In vivo, CAR-Tregs accumulated at sites expressing target antigen and suppressed antigen-specific effector T cell responses; however, only CAR-Tregs with CD28 signaling domains were potent inhibitors of effector T cell–mediated graft rejection in vivo. Our findings support the use of CD28-based CAR-Tregs for tissue-specific immune suppression in the clinic.

Authors

Angela C. Boroughs, Rebecca C. Larson, Bryan D. Choi, Amanda A. Bouffard, Lauren S. Riley, Erik Schiferle, Anupriya S. Kulkarni, Curtis L. Cetrulo, David Ting, Bruce R. Blazar, Shadmehr Demehri, Marcela V. Maus

×

Figure 1

Generation of CAR-Tregs.

Options: View larger image (or click on image) Download as PowerPoint
Generation of CAR-Tregs.
CD4+ T cells were isolated from human donor per...
CD4+ T cells were isolated from human donor peripheral blood mononuclear cells (PBMCs) and enriched for CD25+ cells using positive selection. (A) Sorting gates for Tregs: CD4mid, CD25++, and CD127lo. Sorting gates for Tconvs: CD4+ and CD25lo. Gates were drawn on cells prior to CD25 enrichment. (B) Foxp3 (clone PCH101) intracellular stain after sort. Data are representative of 7 independent sorts from different human donors. (C) Methylation status averaged across sites in the Treg-specific demethylation region (TSDR) on the X chromosome of the sorted T cell populations from female-donor leukopacks (n = 2 female donors, mean plotted). Dots within bars represent individual data points. (D) Vector maps of CD19 CAR constructs. L, leader sequence; scFv, single-chain variable fragment; TM, hinge and transmembrane domain. (E) Experimental design and preparation of CAR-Tregs. (F) Whiskers plots showing mCherry mean fluorescence intensity (MFI) of CAR T cells 12 days after lentivirus transduction at an MOI of 5 measured by flow cytometry (n = 7 human donors). **adj-P < 0.01, by paired ratio t test with Holm-Bonferroni method adjustment for 3 tests between Tregs and Tconvs. Tr, Treg; Tc, Tconv.

Copyright © 2023 American Society for Clinical Investigation
ISSN 2379-3708

Sign up for email alerts