Journal: Science Advances

Article Title: IL-7/IL-15/IL-21 cytokine-fusion scaffold generates highly functional CAR T cells enriched in long-lived T memory stem cells

doi: 10.1126/sciadv.aec2632

Figure Lengend Snippet: ( A ) Schematic of HCW9206. ( B ) Representative histogram plot depicting phospho-STAT5 (pSTAT5) or pSTAT3 expression in CD8 + T cells following HCW9206 (100 nM) stimulation. ( C ) Phospho-STAT5 expression represented by mean fluorescence intensity (MFI) ( N = 3) following stimulation with HCW9206 (100 nM); IL-15, IL-7, and IL-21 (1 nM); or no stimulation. ( D ) Phospho-STAT3 expression represented by MFI ( N = 3). ( E ) Representative histogram plots from one donor (HGLK00122) showing proliferation of human CD8 + T cells and T N/SCM cell memory populations after Cell Trace Violet (CTV) staining following αCD3/28 activation or HCW9206 (100 nM) treatment (T N/SCM cells: CD45RO − CCR7 + ). ( F ) Percentage of proliferating CD8 + T cells following 7-day αCD3/28 stimulation or HCW9206 treatment. Gated on lymphocytes, single cells, CD8 + , and CTV − ( N = 4). ( G ) Percentage of proliferating CD8 + T SCM cells following αCD3/28 stimulation or HCW9206 treatment. Gated on lymphocytes, single cells, CD8 + , CD45RO − , CCR7 + , and CTV − ( N = 4). Data are represented as mean ± SEM. Statistical analysis performed by one-way analysis of variance [ANOVA; (C) and (D)] or unpaired t test [(F) and (G)]. Created in BioRender. E.B.C. (2026). https://BioRender.com/pqy742r .

Article Snippet: T cells were then activated with 1 nM recombinant human IL-7 [National Institutes of Health (NIH)], IL-15 (NIH), or IL-21 (R&D Systems, catalog no. 219-IL-005) cytokines or 100 nM HCW9206 in supplemented IMDM for 7 days before lentiviral transduction.

Techniques: Expressing, Fluorescence, Staining, Activation Assay

Journal: Science Advances

Article Title: IL-7/IL-15/IL-21 cytokine-fusion scaffold generates highly functional CAR T cells enriched in long-lived T memory stem cells

doi: 10.1126/sciadv.aec2632

Figure Lengend Snippet: ( A ) Representation of the CD19-CAR and packaging plasmid maps. ( B ) Percentage of CAR expression in age-matched donor CD8 + T cells after αCD3/28 activation or HCW9206 (100 nM), IL-15, or IL-2 (1 nM) treatment ( N = 3). ( C ) Representative FACS contour plot of the T cell memory phenotype of CD8 + CD19 CAR T cells generated from either αCD3/28 activation or HCW9206 stimulation (left). Histogram plots depicting up-regulation of T SCM cell markers (CD95 and CD45RA) within the HCW9206 CD19 CAR T N/SCM cell population (T N/SCM cells: CD45RO − CCR7 + ) (right). ( D ) CD8 + CD19 CAR T SCM cell frequency in age-matched donors. Gated on lymphocytes, single cells, CD8 + , green fluorescent protein (GFP) + , CD45RO − , CCR7 + , and CD95 + ( N = 3). Statistical analysis compared to HCW9206 treatment. ( E ) CD8 + CD19 CAR T SCM cell frequency comparing IL-7 stimulation (1 nM) to HCW9206 treatment (1 and 100 nM) ( N = 3). ( F ) Progeny analysis depicting frequency changes in CD19 CAR T cell memory populations before and after NALM-6 cell stimulation ( N = 3). Data are shown as the mean ± SEM of triplicates, and statistical analysis was performed using one-way ANOVA [(B) and (D)], one-way repeated-measures ANOVA (E), or two-way ANOVA (F). Created in BioRender. E.B.C. (2026). https://BioRender.com/pqy742r .

Article Snippet: T cells were then activated with 1 nM recombinant human IL-7 [National Institutes of Health (NIH)], IL-15 (NIH), or IL-21 (R&D Systems, catalog no. 219-IL-005) cytokines or 100 nM HCW9206 in supplemented IMDM for 7 days before lentiviral transduction.

Techniques: Plasmid Preparation, Expressing, Activation Assay, Generated, Cell Stimulation