Journal Article

A novel fourth generation of CAR-T cells: CD19 CAR-T cells engineered to express membrane-bound interleukin-15 and CXCR5 for the treatment of lymphoma

Cao, Xinping, Guo, Ruiting, Zhao, Yifan, Liu, Jile, Zhao, Mohan, Guo, Shujing, Zhang, Yi, Zhang, Meng, Zhang, Xiaomei, Sun, Rui, Zhao, Mingfeng

2025

Overview

The efficacy of CD19 CAR-T cells in B-cell lymphoma patients is not as good as that in B-cell acute lymphoblastic leukemia (B-ALL) patients. This might be attributed to the intricate tumor microenvironment of B-cell lymphoma, which leads to CAR-T cell exhaustion, inability to sustain function, and difficulty infiltrating into the tumor interior. We developed CD19 CAR structures that simultaneously produce membrane-bound IL-15 (mbIL-15) and CXCR5. This design aims to enhance the migration of CAR-T cells into CXCL13+ B-cell lymphomas and their long-term antitumor ability. Compared with CD19 CAR-T cells, CD19 mbIL15-CXCR5 CAR-T cells exhibited greater cytotoxicity against CD19+ tumor cell lines in vitro. In particular, when exposed to recurrent tumor antigen stimulation, CD19 mbIL15-CXCR5 CAR-T cells still exerted long-lasting antitumor effects. CD19 mbIL15-CXCR5 CAR-T cells had a greater proportion of central memory T (TCM) and effector memory T (TEM) cells, which allowed them to exhibit more long-lasting antitumor effects. Moreover, in Transwell assays and mouse models, compared with CD19 CAR-T cells, CD19 mbIL15-CXCR5-CAR-T cells exhibited significant chemotaxis toward CXCL13+ tumor cells and superior tumor infiltration ability. The Xenogram animal model demonstrated better and more persistent tumor suppression ability than did the CD19 CAR-T cells. We preliminarily demonstrated the safety of CD19 mbIL15-CXCR5-CAR-T cells in vivo by evaluating liver and kidney function and major organ morphology in mice. In summary, the use of CD19 mbIL15-CXCR5-CAR-T cells is a relatively safe and effective option for the treatment of B-cell malignancies.

• Publisher: Springer International Publishing,Springer Nature B.V
• Subject: Acute lymphoblastic leukemia / Animal models / Animals / Antibodies / Antigens, CD19 - genetics / Antigens, CD19 - immunology / Antigens, CD19 - metabolism / Antitumor activity / B-cell lymphoma / CD19 antigen / Cell Line, Tumor / Cell migration / Cells / Chemokine CXCL13 / Chemokines / Chemotaxis / CXCL13 protein / CXCR5 protein / Cytokines / Cytotoxicity / Effector cells / Female / Flow cytometry / Hematology / Humans / Immunotherapy, Adoptive - methods / Interleukin 15 / Interleukin-15 - genetics / Interleukin-15 - immunology / Interleukin-15 - metabolism / Internal Medicine / Lymphatic leukemia / Lymphocytes / Lymphocytes B / Lymphocytes T / Lymphoma / Lymphoma, B-Cell - immunology / Lymphoma, B-Cell - therapy / Malignancy / Medical research / Medicine / Medicine & Public Health / Memory cells / Metastases / Mice / Oncology / Receptors, Chimeric Antigen - genetics / Receptors, Chimeric Antigen - immunology / Receptors, CXCR5 - genetics / Receptors, CXCR5 - immunology / Receptors, CXCR5 - metabolism / T-Lymphocytes - immunology / Tumor cell lines / Tumor cells / Tumor microenvironment / Tumor suppression / Tumors / Xenograft Model Antitumor Assays