Go to JCI Insight
  • About
  • Editors
  • Consulting Editors
  • For authors
  • Publication ethics
  • Publication alerts by email
  • Advertising
  • Job board
  • Contact
  • Clinical Research and Public Health
  • Current issue
  • Past issues
  • By specialty
    • COVID-19
    • Cardiology
    • Gastroenterology
    • Immunology
    • Metabolism
    • Nephrology
    • Neuroscience
    • Oncology
    • Pulmonology
    • Vascular biology
    • All ...
  • Videos
    • Conversations with Giants in Medicine
    • Video Abstracts
  • Reviews
    • View all reviews ...
    • Pancreatic Cancer (Jul 2025)
    • Complement Biology and Therapeutics (May 2025)
    • Evolving insights into MASLD and MASH pathogenesis and treatment (Apr 2025)
    • Microbiome in Health and Disease (Feb 2025)
    • Substance Use Disorders (Oct 2024)
    • Clonal Hematopoiesis (Oct 2024)
    • Sex Differences in Medicine (Sep 2024)
    • View all review series ...
  • Viewpoint
  • Collections
    • In-Press Preview
    • Clinical Research and Public Health
    • Research Letters
    • Letters to the Editor
    • Editorials
    • Commentaries
    • Editor's notes
    • Reviews
    • Viewpoints
    • 100th anniversary
    • Top read articles

  • Current issue
  • Past issues
  • Specialties
  • Reviews
  • Review series
  • Conversations with Giants in Medicine
  • Video Abstracts
  • In-Press Preview
  • Clinical Research and Public Health
  • Research Letters
  • Letters to the Editor
  • Editorials
  • Commentaries
  • Editor's notes
  • Reviews
  • Viewpoints
  • 100th anniversary
  • Top read articles
  • About
  • Editors
  • Consulting Editors
  • For authors
  • Publication ethics
  • Publication alerts by email
  • Advertising
  • Job board
  • Contact
NOTCH-induced rerouting of endosomal trafficking disables regulatory T cells in vasculitis
Ke Jin, … , Jorg J. Goronzy, Cornelia M. Weyand
Ke Jin, … , Jorg J. Goronzy, Cornelia M. Weyand
Published September 22, 2020
Citation Information: J Clin Invest. 2021;131(1):e136042. https://doi.org/10.1172/JCI136042.
View: Text | PDF
Research Article Autoimmunity Immunology Article has an altmetric score of 4

NOTCH-induced rerouting of endosomal trafficking disables regulatory T cells in vasculitis

  • Text
  • PDF
Abstract

The aorta and the large conductive arteries are immunoprivileged tissues and are protected against inflammatory attack. A breakdown of immunoprivilege leads to autoimmune vasculitis, such as giant cell arteritis, in which CD8+ Treg cells fail to contain CD4+ T cells and macrophages, resulting in the formation of tissue-destructive granulomatous lesions. Here, we report that the molecular defect of malfunctioning CD8+ Treg cells lies in aberrant NOTCH4 signaling that deviates endosomal trafficking and minimizes exosome production. By transcriptionally controlling the profile of RAB GTPases, NOTCH4 signaling restricted vesicular secretion of the enzyme NADPH oxidase 2 (NOX2). Specifically, NOTCH4hiCD8+ Treg cells increased RAB5A and RAB11A expression and suppressed RAB7A, culminating in the accumulation of early and recycling endosomes and sequestering of NOX2 in an intracellular compartment. RAB7AloCD8+ Treg cells failed in the surface translocation and exosomal release of NOX2. NOTCH4hiRAB5AhiRAB7AloRAB11AhiCD8+ Treg cells left adaptive immunity unopposed, enabling a breakdown in tissue tolerance and aggressive vessel wall inflammation. Inhibiting NOTCH4 signaling corrected the defect and protected arteries from inflammatory insult. This study implicates NOTCH4-dependent transcriptional control of RAB proteins and intracellular vesicle trafficking in autoimmune disease and in vascular inflammation.

Authors

Ke Jin, Zhenke Wen, Bowen Wu, Hui Zhang, Jingtao Qiu, Yanan Wang, Kenneth J. Warrington, Gerald J. Berry, Jorg J. Goronzy, Cornelia M. Weyand

×

Figure 5

NOTCH4 signaling regulates the endocytic pathway.

Options: View larger image (or click on image) Download as PowerPoint
NOTCH4 signaling regulates the endocytic pathway.
(A) Confocal microscop...
(A) Confocal microscopy of CD8+ Tregs stained for the early endosome marker EEA1 (green), NOX2 (red), and DAPI (blue). Scale bars: 5 μm. Intensities of EEA1/NOX2 overlap for 10 cells in each group are shown. (B) Confocal microscopy of CD8+ Tregs stained for the recycling endosome marker RAB11 (green), NOX2 (red), and DAPI (blue). Scale bars: 5 μm. Intensities of RAB11/NOX2 overlap in 10 cells from each group are shown. (C and D) CD8+ Tregs were incubated with FITC-labeled transferrin (Tfn) for the indicated times. Internalized Tfn-FITC was determined by flow cytometry. Kinetic measurements (C) for 4 control and 4 GCA Tregs. Tfn uptake at 10 minutes (D) from 9 samples each. (E) CD8+ Tregs were incubated with Tfn-FITC and visualized by confocal microscopy (10 minutes). Scale bars: 10 μm. (F and G) GCA CD8+ Tregs were treated with the endocytosis inhibitor Dynasore (F) or transfected with a RAB5DN-containing or control vector (G). Cell surface NOX2 protein quantified by flow cytometry. Representative histograms and results from 8 patients each. (H) Tfn uptake was quantified in GCA CD8+ Tregs treated with DAPT/vehicle. Flow cytometry data (10 minutes) from 8 samples. Internalized Tfn was visible by confocal microscopy. Scale bars: 10 μm. (I) Tfn uptake was quantified in GCA CD8+ Tregs transfected with NOTCH4 or control siRNA. Flow cytometry data (10 minutes) from 8 patients. (J) Tfn uptake was quantified by FACS in healthy CD8+ Tregs transfected with a NOTCH4 ICD-containing or empty vector. Results from 10 samples. (K) HC CD8+ Tregs transfected with empty vector or NOTCH4 ICD-containing plasmid were treated with Dynasore or vehicle. Cell surface NOX2 protein was determined by flow cytometry. Representative histograms and results from 9 samples. CD8+ Treg cells were induced ex vivo. Data are mean ± SEM. *P < 0.05; **P < 0.01; ***P < 0.001 by unpaired Mann-Whitney-Wilcoxon rank test (A, B, and D), paired Mann-Whitney-Wilcoxon rank test (E–I), or 1-way ANOVA and post-ANOVA pairwise 2-group comparisons conducted with Tukey’s method (J).

Copyright © 2025 American Society for Clinical Investigation
ISSN: 0021-9738 (print), 1558-8238 (online)

Sign up for email alerts

Posted by 8 X users
31 readers on Mendeley
See more details