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Phenotypic characterisation of expanded natural killer (NK) cells. NK cells were characterised on Day 14 of expansion. Uniform Manifold Approximation and Projection (UMAP) plots show the 10 distinct meta clusters of NK cells generated via FLOWSOM (a) , with a contour UMAP indicating proportions of NK cells belonging to each cluster (b) . A clustered heatmap visualisation displays the MFI of each indicated marker belonging to NK cells within the corresponding clusters (c) . To model <t>ADCC,</t> <t>PD‐L1</t> was over‐expressed on MCF‐7 cells (d) prior to cytotoxicity assay in the presence/absence of 10 μg/mL avelumab (e) . NK Cell phenotyping, n = 3. Cytotoxicity displayed as individual donors ( n = 4). ** P ≤ 0.005, by multiple paired t ‐tests.
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Phenotypic characterisation of expanded natural killer (NK) cells. NK cells were characterised on Day 14 of expansion. Uniform Manifold Approximation and Projection (UMAP) plots show the 10 distinct meta clusters of NK cells generated via FLOWSOM (a) , with a contour UMAP indicating proportions of NK cells belonging to each cluster (b) . A clustered heatmap visualisation displays the MFI of each indicated marker belonging to NK cells within the corresponding clusters (c) . To model <t>ADCC,</t> <t>PD‐L1</t> was over‐expressed on MCF‐7 cells (d) prior to cytotoxicity assay in the presence/absence of 10 μg/mL avelumab (e) . NK Cell phenotyping, n = 3. Cytotoxicity displayed as individual donors ( n = 4). ** P ≤ 0.005, by multiple paired t ‐tests.
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Phenotypic characterisation of expanded natural killer (NK) cells. NK cells were characterised on Day 14 of expansion. Uniform Manifold Approximation and Projection (UMAP) plots show the 10 distinct meta clusters of NK cells generated via FLOWSOM (a) , with a contour UMAP indicating proportions of NK cells belonging to each cluster (b) . A clustered heatmap visualisation displays the MFI of each indicated marker belonging to NK cells within the corresponding clusters (c) . To model <t>ADCC,</t> <t>PD‐L1</t> was over‐expressed on MCF‐7 cells (d) prior to cytotoxicity assay in the presence/absence of 10 μg/mL avelumab (e) . NK Cell phenotyping, n = 3. Cytotoxicity displayed as individual donors ( n = 4). ** P ≤ 0.005, by multiple paired t ‐tests.
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Phenotypic characterisation of expanded natural killer (NK) cells. NK cells were characterised on Day 14 of expansion. Uniform Manifold Approximation and Projection (UMAP) plots show the 10 distinct meta clusters of NK cells generated via FLOWSOM (a) , with a contour UMAP indicating proportions of NK cells belonging to each cluster (b) . A clustered heatmap visualisation displays the MFI of each indicated marker belonging to NK cells within the corresponding clusters (c) . To model <t>ADCC,</t> <t>PD‐L1</t> was over‐expressed on MCF‐7 cells (d) prior to cytotoxicity assay in the presence/absence of 10 μg/mL avelumab (e) . NK Cell phenotyping, n = 3. Cytotoxicity displayed as individual donors ( n = 4). ** P ≤ 0.005, by multiple paired t ‐tests.
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Phenotypic characterisation of expanded natural killer (NK) cells. NK cells were characterised on Day 14 of expansion. Uniform Manifold Approximation and Projection (UMAP) plots show the 10 distinct meta clusters of NK cells generated via FLOWSOM (a) , with a contour UMAP indicating proportions of NK cells belonging to each cluster (b) . A clustered heatmap visualisation displays the MFI of each indicated marker belonging to NK cells within the corresponding clusters (c) . To model <t>ADCC,</t> <t>PD‐L1</t> was over‐expressed on MCF‐7 cells (d) prior to cytotoxicity assay in the presence/absence of 10 μg/mL avelumab (e) . NK Cell phenotyping, n = 3. Cytotoxicity displayed as individual donors ( n = 4). ** P ≤ 0.005, by multiple paired t ‐tests.
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Phenotypic characterisation of expanded natural killer (NK) cells. NK cells were characterised on Day 14 of expansion. Uniform Manifold Approximation and Projection (UMAP) plots show the 10 distinct meta clusters of NK cells generated via FLOWSOM (a) , with a contour UMAP indicating proportions of NK cells belonging to each cluster (b) . A clustered heatmap visualisation displays the MFI of each indicated marker belonging to NK cells within the corresponding clusters (c) . To model <t>ADCC,</t> <t>PD‐L1</t> was over‐expressed on MCF‐7 cells (d) prior to cytotoxicity assay in the presence/absence of 10 μg/mL avelumab (e) . NK Cell phenotyping, n = 3. Cytotoxicity displayed as individual donors ( n = 4). ** P ≤ 0.005, by multiple paired t ‐tests.
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Phenotypic characterisation of expanded natural killer (NK) cells. NK cells were characterised on Day 14 of expansion. Uniform Manifold Approximation and Projection (UMAP) plots show the 10 distinct meta clusters of NK cells generated via FLOWSOM (a) , with a contour UMAP indicating proportions of NK cells belonging to each cluster (b) . A clustered heatmap visualisation displays the MFI of each indicated marker belonging to NK cells within the corresponding clusters (c) . To model ADCC, PD‐L1 was over‐expressed on MCF‐7 cells (d) prior to cytotoxicity assay in the presence/absence of 10 μg/mL avelumab (e) . NK Cell phenotyping, n = 3. Cytotoxicity displayed as individual donors ( n = 4). ** P ≤ 0.005, by multiple paired t ‐tests.

Journal: Clinical & Translational Immunology

Article Title: Parallel processing of T cells and natural killer cells to enhance in vivo CAR T cell activity against blood and solid cancers

doi: 10.1002/cti2.70120

Figure Lengend Snippet: Phenotypic characterisation of expanded natural killer (NK) cells. NK cells were characterised on Day 14 of expansion. Uniform Manifold Approximation and Projection (UMAP) plots show the 10 distinct meta clusters of NK cells generated via FLOWSOM (a) , with a contour UMAP indicating proportions of NK cells belonging to each cluster (b) . A clustered heatmap visualisation displays the MFI of each indicated marker belonging to NK cells within the corresponding clusters (c) . To model ADCC, PD‐L1 was over‐expressed on MCF‐7 cells (d) prior to cytotoxicity assay in the presence/absence of 10 μg/mL avelumab (e) . NK Cell phenotyping, n = 3. Cytotoxicity displayed as individual donors ( n = 4). ** P ≤ 0.005, by multiple paired t ‐tests.

Article Snippet: The human PD‐L1 (NCBI 29126) gene with endogenous signal sequence was synthesised by Twist Biosciences and cloned into SfiI sites of pSBbiP—a modified pSBbiGP encoding puromycin resistance but with the original GFP gene removed.

Techniques: Generated, Marker, Cytotoxicity Assay

Recycled natural killer (NK) cells enhance CAR T cell therapy of MCF‐7. NSG mice were engrafted with Her2 h i PDL1 hi MCF‐7 cells on Day 0 prior to sequential administration of NK cells (d10) and CAR T cells (d12) (a) . Quality control of both cell types ensured high NK cell purity and transduction efficiency of T cells prior to administration (b, c) . Mice were imaged weekly (d, e) , and callipers were used every other day to monitor tumor burden (f) . Mice were euthanised when radiance and/or tumor volume reached endpoint thresholds (g) . Data from one mouse experiment. n = 4–5 mice per group. Tumor burdens displayed as mean ± SEM. * P < 0.05 by two‐way ANOVA with Bonferroni correction. Survival analysed with Kaplan–Meier, log‐rank mantel‐cox test. ** P ≤ 0.005. White out‐lined box conceals mouse removed from study because of failed tumor engraftment.

Journal: Clinical & Translational Immunology

Article Title: Parallel processing of T cells and natural killer cells to enhance in vivo CAR T cell activity against blood and solid cancers

doi: 10.1002/cti2.70120

Figure Lengend Snippet: Recycled natural killer (NK) cells enhance CAR T cell therapy of MCF‐7. NSG mice were engrafted with Her2 h i PDL1 hi MCF‐7 cells on Day 0 prior to sequential administration of NK cells (d10) and CAR T cells (d12) (a) . Quality control of both cell types ensured high NK cell purity and transduction efficiency of T cells prior to administration (b, c) . Mice were imaged weekly (d, e) , and callipers were used every other day to monitor tumor burden (f) . Mice were euthanised when radiance and/or tumor volume reached endpoint thresholds (g) . Data from one mouse experiment. n = 4–5 mice per group. Tumor burdens displayed as mean ± SEM. * P < 0.05 by two‐way ANOVA with Bonferroni correction. Survival analysed with Kaplan–Meier, log‐rank mantel‐cox test. ** P ≤ 0.005. White out‐lined box conceals mouse removed from study because of failed tumor engraftment.

Article Snippet: The human PD‐L1 (NCBI 29126) gene with endogenous signal sequence was synthesised by Twist Biosciences and cloned into SfiI sites of pSBbiP—a modified pSBbiGP encoding puromycin resistance but with the original GFP gene removed.

Techniques: Control, Transduction