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
Enhancing CAR T cell persistence through ICOS and 4-1BB costimulation
Sonia Guedan, … , Regina M. Young, Carl H. June
Sonia Guedan, … , Regina M. Young, Carl H. June
Published January 11, 2018
Citation Information: JCI Insight. 2018;3(1):e96976. https://doi.org/10.1172/jci.insight.96976.
View: Text | PDF
Research Article Immunology

Enhancing CAR T cell persistence through ICOS and 4-1BB costimulation

  • Text
  • PDF
Abstract

Successful tumor eradication by chimeric antigen receptor–expressing (CAR-expressing) T lymphocytes depends on CAR T cell persistence and effector function. We hypothesized that CD4+ and CD8+ T cells may exhibit distinct persistence and effector phenotypes, depending on the identity of specific intracellular signaling domains (ICDs) used to generate the CAR. First, we demonstrate that the ICOS ICD dramatically enhanced the in vivo persistence of CAR-expressing CD4+ T cells that, in turn, increased the persistence of CD8+ T cells expressing either CD28- or 4-1BB–based CARs. These data indicate that persistence of CD8+ T cells was highly dependent on a helper effect provided by the ICD used to redirect CD4+ T cells. Second, we discovered that combining ICOS and 4-1BB ICDs in a third-generation CAR displayed superior antitumor effects and increased persistence in vivo. Interestingly, we found that the membrane-proximal ICD displayed a dominant effect over the distal domain in third-generation CARs. The optimal antitumor and persistence benefits observed in third-generation ICOSBBz CAR T cells required the ICOS ICD to be positioned proximal to the cell membrane and linked to the ICOS transmembrane domain. Thus, CARs with ICOS and 4-1BB ICD demonstrate increased efficacy in solid tumor models over our current 4-1BB–based CAR and are promising therapeutics for clinical testing.

Authors

Sonia Guedan, Avery D. Posey Jr., Carolyn Shaw, Anna Wing, Tong Da, Prachi R. Patel, Shannon E. McGettigan, Victoria Casado-Medrano, Omkar U. Kawalekar, Mireia Uribe-Herranz, Decheng Song, J. Joseph Melenhorst, Simon F. Lacey, John Scholler, Brian Keith, Regina M. Young, Carl H. June

×

Figure 1

In vitro characterization of CD4+ and CD8+ T cells redirected with SS1-CARs.

Options: View larger image (or click on image) Download as PowerPoint
In vitro characterization of CD4+ and CD8+ T cells redirected with SS1-C...
(A) Schematic representation of chimeric receptors that contain the SS1 single chain fragment that binds to mesothelin and differ in the transmembrane and intracellular domains. (B) Surface expression of the SS1-CARs on human CD4+ T cells at the end of the primary expansion. Representative of 3 donors. (C) A real-time, impedance-based cytotoxicity assay (xCelligence) was used to evaluate the lysis of non–small cell lung L55 tumor cells when treated with CAR T cells at 1:1 E:T ratio over a 20-hour period. Representative of 2 donors. (D) CD4+ CAR T cells were cocultured with pancreatic cancer cells (Capan-2) that express mesothelin. Supernatants were obtained 24 hours after coculture, and cytokine production was analyzed by Luminex. Representative of 3 different experiments performed under similar conditions. (E) Ratios of cytokine expression for 28z and ICOSz were calculated using results from C. (F) CD4+ CAR T cells from different donors (n = 3–6) were cocultured with APC cells modified to express mesothelin. Supernatants were obtained 24 hours after coculture, and cytokine production was analyzed by ELISA. Ratios of cytokine expression for 28z and ICOSz are shown. Box plots show median (line) and 25th to 75th percentile (box). The end of the whiskers represents the minimum and the maximum of all of the data. *P < 0.05 by t-test. (G) CD4+ CAR T cells were cocultured with Capan-2 or immobilized mesothelin. Supernatants were obtained 24 hours after coculture, and IL-6 production was analyzed by Luminex. Representative of 3 donors (H) CD4+ CAR T cells were stimulated with magnetic beads coated with recombinant mesothelin. Cell lysates were obtained at different time points (5, 10, 30 and 60 min) and phosphorylation levels for AKT and ERK were analyzed by Western Blot and densitometry. Basal phosphorylation was evaluated without stimulation (minute 0). Representative of 3 donors.

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

Sign up for email alerts