Preclinical comparison of non-signaling domain in CD19 CAR T cell with interleukin-7 receptor alpha signaling domain

Pornlapat Keawvilai; Sirirut Jewmoung; Kristine Cate S. Pe; Supannikar Tawinwung; Koramit Suppipat

doi:10.1016/j.biopha.2026.119220

Preclinical comparison of non-signaling domain in CD19 CAR T cell with interleukin-7 receptor alpha signaling domain

Pornlapat Keawvilai, Sirirut Jewmoung, Kristine Cate S. Pe, Supannikar Tawinwung Koramit Suppipat

Biomedicine & pharmacotherapy, Vol.198, 119220

18/03/2026

: https://doi.org/10.1016/j.biopha.2026.119220

https://hdl.handle.net/10523/50365

B-cell malignancies

CAR T cell

CD19

IL-7 receptor alpha (IL-7Rα)

Non-signaling domain

Chimeric antigen receptor (CAR) T cells have emerged as an effective immunotherapy for hematologic malignancies. The non-signaling domain of CARs, comprising the spacer and transmembrane regions, is a key structural component that can be engineered to influence CAR expression and function. In this study, we evaluated three non-signaling domain configurations—IgG2.CH3/CD28, IgG2/CD28, and CD8/CD8—within a CD19 CAR construct incorporating 4–1BB and interleukin-7 receptor alpha (IL-7Rα) signaling domains. CARs incorporating the IgG2/CD28 domain exhibited reduced surface expression and diminished functional responses compared with IgG2.CH3/CD28 and CD8/CD8 constructs. The CD8/CD8 configuration supported the highest CAR expression and sustained surface density. In contrast, IgG2.CH3/CD28 CAR T cells displayed increased IL-2 and TNF-α secretion and enhanced CD107α upregulation following antigen stimulation. In a serial tumor cell rechallenge assay, IgG2.CH3/CD28 CAR T cells maintained cytotoxic activity and persistence compared with CD8/CD8 CAR T cells. In a NALM-6 xenograft model, IgG2.CH3/CD28 CAR T cells achieved durable tumor control and were associated with improved survival relative to CD8/CD8 CAR T cells. Collectively, these findings support the IgG2.CH3/CD28 non-signaling domain as a suitable structural component for CD19 CARs incorporating IL-7Rα signaling and provide insight into CAR design strategies aimed at improving T cell persistence and anti-leukemic activity.

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https://doi.org/10.1016/j.biopha.2026.119220

Published (Version of record),

: 9926853280201891
: Preclinical comparison of non-signaling domain in CD19 CAR T cell with interleukin-7 receptor alpha signaling domain
: Pornlapat Keawvilai
Sirirut Jewmoung
Kristine Cate S. Pe
Supannikar Tawinwung
Koramit Suppipat
: Microbiology and Immunology
: Biomedicine & pharmacotherapy, Vol.198, 119220
: Elsevier
: 18/03/2026
: Ratchadapisek Somphot Fund for Postdoctoral Fellowship, Chulalongkorn University; Thailand Science Research and Innovation Fund Chulalongkorn University (HEA_FF_69_231_3000_032); National Research Council of Thailand and Chulalongkorn University (N42A680237)
: Copyright © The Author(s) 2026. This work was first published in Biomedicine & Pharmacotherapy (Elsevier). This is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International License (https://www.creativecommons.org/licenses/by/4.0/), which permits unrestricted use, sharing, adaptation, distribution and reproduction in any medium or format, provided that the original work is properly attributed to the creator(s) and the source, a link to the Creative Commons license is provided, and any changes made are indicated.
: English
: Journal article