TY - JOUR
AU - Moser, Laura M
AU - Heim, Catrin
AU - Koschade, Sebastian E
AU - Wendel, Philipp
AU - Bozkurt, Süleyman
AU - Harenkamp, Sabine
AU - Kreyenberg, Hermann
AU - Merker, Michael
AU - Münch, Christian
AU - Gradhand, Elise
AU - Vogler, Meike
AU - Ullrich, Evelyn
AU - Bönig, Halvard
AU - Klusmann, Jan-Henning
AU - Bader, Peter
AU - Wels, Winfried S
AU - Rettinger, Eva
TI - CAR-CIK vs. CAR-T: benchmarking novel cytokine-induced killer cells as solid tumor immunotherapy in ErbB2+ rhabdomyosarcoma.
JO - Frontiers in immunology
VL - 16
SN - 1664-3224
CY - Lausanne
PB - Frontiers Media
M1 - DKFZ-2025-00382
SP - 1485817
PY - 2025
AB - CAR-T cell therapy, though successful in hematologic malignancies, faces challenges in solid tumors due to limitations of autologous T cells. Cytokine-induced killer (CIK) cells can be given safely across allogeneic barriers and constitute alternative effector cells generated from healthy donors. CIK cells are a heterogenous population of predominantly T cells with a mixed natural killer (NK) phenotype and combine non-MHC-restricted cytotoxicity with potent anti-tumor capacity of the adaptive immune system. Here, we characterize and compare efficacy, phenotypic subpopulations and modes of action of CAR-CIK cells and conventional CAR-T cells from same-donor samples in ErbB2+ rhabdomyosarcoma (RMS).To benchmark CAR-CIK against conventional CAR-T cells, effector cells were generated from same-donor samples and lentivirally transduced with a second generation CD28-CD3ζ CAR. Effector subpopulations and their dynamics upon target cell exposure were phenotypically characterized by flow cytometry. Efficacy was assessed in human ErbB2+ RMS cancer cell lines and primary patient samples in vitro and ex vivo using cytotoxicity and spheroid co-incubation assays. Modes of action were assessed by comparing cytokine secretion profiles using bead-based multiplexed flow cytometry and by liquid chromatography mass spectrometry whole cell proteomics. Finally, we used an in vivo model of RMS mimicking minimal metastatic residual disease to compare anti-tumor potency of CAR-CIK vs. CAR-T cells and to assess their target organ infiltration.In vitro assays demonstrated superior cytotoxicity of CAR-CIK cells against RMS cell lines and primary tumor samples. Long-term co-incubation with tumor spheroids led to expansion of CAR-CIK cells and enrichment of CD3+CD56+ TNK cells. CAR-CIK cell cytokine signature showed significantly increased secretion of effector molecules like interferon-γ, perforin and granulysin, and lower secretion of Th2 cytokines IL-2, IL-4 and IL-10. Whole cell proteomics showed corresponding upregulation of chemokine signaling and NK-cytotoxicity pathways in CAR-CIK cells. In NSG mice xenografted with ErbB2+ RMS, a single injection of either CAR-effector cells strongly impeded metastatic tumor development and significantly improved survival.Our results demonstrate that CAR-CIK cells are at least equipotent to CAR-T cells. Combined with their favorable safety profile and allogeneic applicability, these findings position CAR-CIK cells as promising immune effectors for solid tumors.
KW - Humans
KW - Rhabdomyosarcoma: therapy
KW - Rhabdomyosarcoma: immunology
KW - Cytokine-Induced Killer Cells: immunology
KW - Immunotherapy, Adoptive: methods
KW - Animals
KW - Receptor, ErbB-2: immunology
KW - Receptors, Chimeric Antigen: immunology
KW - Receptors, Chimeric Antigen: genetics
KW - Receptors, Chimeric Antigen: metabolism
KW - Mice
KW - Cell Line, Tumor
KW - Xenograft Model Antitumor Assays
KW - Cytokines: metabolism
KW - Cytotoxicity, Immunologic
KW - Benchmarking
KW - CAR-T (Other)
KW - ERBB2 (Other)
KW - cytokine-induced killer cells (CIK) (Other)
KW - rhabdomyosarcoma (Other)
KW - solid tumors (Other)
KW - Receptor, ErbB-2 (NLM Chemicals)
KW - Receptors, Chimeric Antigen (NLM Chemicals)
KW - ERBB2 protein, human (NLM Chemicals)
KW - Cytokines (NLM Chemicals)
LB - PUB:(DE-HGF)16
C6 - pmid:39963129
C2 - pmc:PMC11831232
DO - DOI:10.3389/fimmu.2025.1485817
UR - https://inrepo02.dkfz.de/record/298948
ER -