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. 2023 Jul 5;7(20):6009–6022. doi: 10.1182/bloodadvances.2023010032

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© 2023 by The American Society of Hematology. Licensed under Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0), permitting only noncommercial, nonderivative use with attribution. All other rights reserved.

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Figure 2. — Anti-tumor efficacy of BCMA CAR-iNKTs is enhanced with rhIL-7-hyFc. (A) In vivo schema. Actively growing BCMA CAR-Ts, CD19 CAR-Ts, BCMA CAR-iNKTs and CD19 CAR-iNKTs were used in this experiment. rhIL-7-hyFc (10 mg/kg; NeoImmuneTech, Inc) was injected subcutaneously on days +1 and +14 after CAR-iNKT administration. (B) Kaplan-Meier survival curve. (C) Normalized longitudinal BLI images. (D) Quantitation of photon flux. (E) Absolute number of CAR-iNKTs per μL blood. Because the BCMA CAR-iNKTs were 100% CD4⁺ (not shown), we initially used CD45, CD3, CD4, and CD34 to track CAR⁺ iNKTs in vivo. On day 64, the panel was expanded and confirmed that all CD4⁺ cells were Vα24-Jα18⁺. (F) In vivo schema. Previously expanded and cryopreserved NTD and M-BB BCMA CAR-iNKTs were used in this experiment. Cells were thawed the day before injection and placed in the incubator overnight, counted, and injected into mice with MM1S-CG engraftment. (G) Kaplan-Meier survival analysis. (H) Longitudinal normalized BLI images. (I) Quantitation of BLI signal. (J) Absolute number of CAR-iNKTs per μL blood (CD45, CD3, Vα24-Jα18, and CD34). On day 52, total CD45⁺ cells are shown because of a technical issue.

Figure 2. — Anti-tumor efficacy of BCMA CAR-iNKTs is enhanced with rhIL-7-hyFc. (A) In vivo schema. Actively growing BCMA CAR-Ts, CD19 CAR-Ts, BCMA CAR-iNKTs and CD19 CAR-iNKTs were used in this experiment. rhIL-7-hyFc (10 mg/kg; NeoImmuneTech, Inc) was injected subcutaneously on days +1 and +14 after CAR-iNKT administration. (B) Kaplan-Meier survival curve. (C) Normalized longitudinal BLI images. (D) Quantitation of photon flux. (E) Absolute number of CAR-iNKTs per μL blood. Because the BCMA CAR-iNKTs were 100% CD4⁺ (not shown), we initially used CD45, CD3, CD4, and CD34 to track CAR⁺ iNKTs in vivo. On day 64, the panel was expanded and confirmed that all CD4⁺ cells were Vα24-Jα18⁺. (F) In vivo schema. Previously expanded and cryopreserved NTD and M-BB BCMA CAR-iNKTs were used in this experiment. Cells were thawed the day before injection and placed in the incubator overnight, counted, and injected into mice with MM1S-CG engraftment. (G) Kaplan-Meier survival analysis. (H) Longitudinal normalized BLI images. (I) Quantitation of BLI signal. (J) Absolute number of CAR-iNKTs per μL blood (CD45, CD3, Vα24-Jα18, and CD34). On day 52, total CD45⁺ cells are shown because of a technical issue.