CAR T Cell-Induced Interferon Gamma Enhances MHC Class I Expression and Sensitizes Neuroblastoma to TCR-Engineered T Cell Therapy

Authors

• Alice Vitali
• Jennie Padlo
• Carlotta Caramel
• Lena Andersch
• Laura M Hartmann
• Jonas Kath
• Laura Grunewald
• Silke Schwiebert
• Anika Winkler
• Efrosiinia Talalai
• Tabea Biereder
• Judith Wienke
• Jan J Molenaar
• Michael Launspach
• Catharina Delebinski
• Anton Kern
• Kathy Astrahantseff
• Dimitrios L Wagner
• Gerald Willimsky
• Angelika Eggert
• Thomas Blankenstein
• Kathleen Anders
• Annette Künkele

Language

English

Publication Date

1-1-2026

Journal

Frontiers in Immunology

DOI

10.3389/fimmu.2026.1901504

PMID

42827984

PMCID

PMC13631103

PubMedCentral® Posted Date

9-18-2026

PubMedCentral® Full Text Version

Post-print

Abstract

Background: T cell receptor (TCR)-engineered T cells target antigens presented on major histocompatibility complex (MHC) class I molecules. MHC class I expression is often low in childhood tumors such as neuroblastoma but can be stimulated by interferon gamma (IFNγ). Chimeric antigen receptor (CAR) T cell effector function, including IFNγ production, is MHC class I-independent.

Aim: We aimed to enhance neuroblastoma immunotherapy by combining CAR- and TCR-engineered T cells. Our strategy exploits CAR T cell-derived effector molecules, including IFNγ, to increase tumor cell MHC class I expression and thereby enhance subsequent TCR T cell activity.

Methods: IFNγ-mediated induction and persistence of MHC class I expression were analyzed in neuroblastoma cell lines by flow cytometry. Primary human T cells were engineered to express either a high-affinity TCR targeting the cancer testis antigen NY-ESO-1 or an L1CAM-targeted CAR. Cytotoxic activity of CAR and TCR T cells, applied alone or sequentially, was quantified against LAN-1 neuroblastoma cells using IncuCyte-based real-time killing assays. To model heterogeneous antigen expression, mixed tumor populations containing L1CAM-positive and CRISPR/Cas9-generated L1CAM-knockout cells were analyzed. In vivo MHC class I upregulation was assessed in a human tumor xenograft model following treatment with human L1CAM-CAR T cells and in a syngeneic tumor model following treatment with GD2-CAR T cells.

Results: NY-ESO-1 TCR T cells alone showed limited cytotoxicity against LAN-1 cells in vitro, whereas L1CAM-CAR T cell pretreatment increased MHC class I expression and enhanced subsequent TCR-mediated killing. In tumor cells with heterogeneous CAR antigen expression, CAR T cell recognition of antigen-positive cells induced HLA-A*02 upregulation across the tumor population, enabling subsequent NY-ESO-1 TCR T cell recognition of cells not directly recognized by CAR T cells. Exploratory in vivo analyses further showed increased tumor MHC class I expression following CAR T cell treatment, providing preliminary evidence of CAR T cell-mediated inflammatory priming in vivo.

Conclusion: Sequential CAR- and TCR-engineered T cell therapy may help overcome limitations imposed by low MHC class I and heterogeneous antigen expression in neuroblastoma, supporting further investigation of this combinatorial strategy.

Keywords

Humans, Neuroblastoma, Animals, Interferon-gamma, Immunotherapy, Adoptive, Receptors, Chimeric Antigen, Cell Line, Tumor, Histocompatibility Antigens Class I, Mice, T-Lymphocytes, Receptors, Antigen, T-Cell, Xenograft Model Antitumor Assays, Cytotoxicity, Immunologic, antigen loss, MHC class I low, neuroblastoma, T cell therapy, TCR T cell resistance, tumor heterogeneity

Published Open-Access

yes

Recommended Citation

Vitali, Alice; Padlo, Jennie; Caramel, Carlotta; et al., "CAR T Cell-Induced Interferon Gamma Enhances MHC Class I Expression and Sensitizes Neuroblastoma to TCR-Engineered T Cell Therapy" (2026). Faculty, Staff and Students Publications. 8687.
https://digitalcommons.library.tmc.edu/baylor_docs/8687