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. 2024 Jan 2;15:89. doi: 10.1038/s41467-023-44310-y

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© The Author(s) 2024, corrected publication 2024

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 7 — a Schematic of the retroviral vector encoding the GD2.CAR and membrane-bound IL-12 in which the mutated CD8α stalk was added to IL-12 (IL12M). b Representative flow cytometry plots of 5 donors showing CAR expression in GD2.CAR(I)IL12 and GD2.CAR(I)IL12.TM NKTs assessed at day 14 of culture. Gray line represents non-transduced NKTs. c Representative flow cytometry plots of 5 donors showing the expression of IL-12 on the cell surface of GD2.CAR(I)IL12 and GD2.CAR(I)IL12.TM NKTs assessed at day 14 of culture. Gray line represents non-transduced NKTs. d Representative flow cytometry plots of 5 donors showing CD62L expression in GD2.CAR(I)IL12 and GD2.CAR(I)IL12.TM NKTs assessed at day 14 of culture. e Representative western blot illustrating STAT4 phosphorylation in NT, CD19.CAR(I)IL12, GD2.CAR, GD2.CAR(I)IL12 and GD2.CAR(I)IL12.TM NKTs at day 14 of culture; n = 3. Source data are provided as a Source Data file. f Schematic representation of the metastatic xenograft neuroblastoma model. Mice were engrafted i.v. with 1 × 10⁶ CHLA-255 Firefly-luciferase labeled neuroblastoma tumor cells and treated with 5 × 106 NT, GD2.CAR, GD2.CAR(I)IL12 and GD2.CAR(I)IL12.TM NKTs. g Representative tumor BLI; n = 4 from two independent experiments. h Kaplan–Meier survival curve showing overall survival; n = 4 mice, *p = 0.0344 GD2.CAR vs GD2.CAR(I)IL12.TM NKTs; log-rank Mantel–Cox test. Source data are provided as a Source Data file. i Representative flow cytometry plots (left panels) and quantification of human NKTs (iNKT⁺CD45⁺) in peripheral blood samples (right panel) collected 4 weeks after NKT infusion. Mean is shown; n = 4 mice; **p = 0.0060 GD2.CAR vs GD2.CAR(I)IL12 NKTs, **p = 0.0035 GD2.CAR vs GD2.CAR(I)IL12.TM NKTs, two-tailed unpaired t test. Source data are provided as a Source Data file. j Quantification of IL-12 and IFN-γ detected in the serum of mice at sacrifice. Mean is shown; n = 4 mice; IL-12: **p = 0.0077 GD2.CAR vs GD2.CAR(I)IL12, **p = 0.0081 GD2.CAR(I)IL12 vs GD2.CAR(I)IL12.TM; IFNγ: **p = 0.0032 GD2.CAR vs GD2.CAR(I)IL12, **p = 0.0086 GD2.CAR vs GD2.CAR(I)IL12.TM; two-tailed paired t test. Source data are provided as a Source Data file. k, L Metastatic xenograft neuroblastoma model in which mice were treated with GD2.CAR.IL15 or GD2.CAR(I)IL12 or GD2.CAR(I)IL12.TM NKTs following the tumor model described in (f). k Representative tumor BLI. L Kaplan–Meier survival curve showing overall survival; n = 3 mice for NT, n = 4 mice for GD2.CAR(I)IL12 and n = 5 mice for GD2.CAR.IL15 and GD2.CAR(I)IL12.TM NKTs, *p = 0.0112 GD2.CAR.IL15 vs GD2.CAR(I)IL12; *p = 0.0197 GD2.CAR.IL15 vs GD2.CAR(I)IL12.TM NKTs; log-rank Mantel–Cox test. Source data are provided as a Source Data file.