Journal: eBioMedicine

Article Title: Tandem CD19/CD22 CAR T-cells as potential therapy for children and young adults with high-risk r/r B-ALL

doi: 10.1016/j.ebiom.2025.105872

Figure Lengend Snippet: Tandem anti-CD19/CD22 CAR T-cell therapy response. A) Schematic diagram of the tandem anti-CD19/CD22 CAR structure. B) Flow chart of the study. C) Swimmer plot showing clinical responses after tandem anti-CD19/CD22 CAR T-cell product infusion. D) PET-TC imaging of patient P9 before (A.1 and A.2) and 28 days after (B.1 and B.2) tandem anti-CD19/CD22 CAR T-cell infusion. E) Event free survival (EFS) Kaplan–Meier curve in all patients (n = 10). F) Overall survival (OS) Kaplan–Meier curve in all patients (n = 10). For E-F, black dots on the curve represent censored observations. HSCT, haematopoietic stem cell transplantation. MRD, minimal residual disease. EMR, extramedullary relapse. PD, progression of disease. ∗ For P6, 6 reinfusions were performed (every two weeks), the last with 3 doses. ∗∗P4 joined a clinical trial with carfilzomib after relapse, with PD shortly after.

Article Snippet: The CD19 (130-129-550) and CD22 (130-126-727) CAR detection reagents, anti-CD19-Viogreen (130-113-649), anti-CD3-Viogreen (130-113-142), and anti-biotin-PE (130-110-951) were from Miltenyi Biotec.

Techniques: Imaging, Transplantation Assay

Journal: eBioMedicine

Article Title: Tandem CD19/CD22 CAR T-cells as potential therapy for children and young adults with high-risk r/r B-ALL

doi: 10.1016/j.ebiom.2025.105872

Figure Lengend Snippet: Tandem anti-CD19/CD22 CAR T-cell product immunophenotype and analysis. A) Tandem anti-CD19/CD22 CAR T-cell expansion in all products manufactured in the CliniMACS Prodigy closed system. B) CD19-CAR and CD22-CAR expression in all products manufactured by flow cytometry. C) CD4 + and CD8 + cell populations in all 10 products manufactured. D) PD-1 receptor expression on product cells. In C and D, graphs show box and whisker plots (vertical bars, min to max points; box, first to third quartile, with median as horizontal bar). E) Memory subpopulations determined by flow cytometry. Central memory cells (CD45RA − CD27 + ), effector memory cells (CD45RA − CD27 − ), naïve cells (CD45RA + CD27 + ) and TEMRA cells (CD45RA + CD27 − ) are represented. F) Specific-lysis by tandem anti-CD19/CD22 CAR T-cells against SEM cell line determined by 4-h Europium-BATDA assay at different E:T ratios. G) Degranulation assay against SEM cell line determined by CD107a expression after 4 h of co-culture at 1:1 or 1:2 E:T ratios (see Methods). A–D and F and G , in green, living patients; in purple, relapsed patient; in black, patients who died.

Article Snippet: The CD19 (130-129-550) and CD22 (130-126-727) CAR detection reagents, anti-CD19-Viogreen (130-113-649), anti-CD3-Viogreen (130-113-142), and anti-biotin-PE (130-110-951) were from Miltenyi Biotec.

Techniques: Expressing, Flow Cytometry, Whisker Assay, Lysis, Degranulation Assay, Co-Culture Assay

Journal: eBioMedicine

Article Title: Tandem CD19/CD22 CAR T-cells as potential therapy for children and young adults with high-risk r/r B-ALL

doi: 10.1016/j.ebiom.2025.105872

Figure Lengend Snippet: Tandem anti-CD19/CD22 CAR T-cell persistence in patients after product infusion. A) Tandem anti-CD19/CD22 CAR expression was determined with anti-CD19 CAR gated on CD3 + cells. Left panel, absolute numbers of peripheral blood CAR T-cells per μl in infused patients detected by flow cytometry. Right panel, percentage of CAR + cells within the T cell compartment. B) Copies per ml as detected using real-time qPCR (see Methods) (16). C) IL-6 levels from serum after CAR T-cell infusion. D) Peak IL-6 levels were upregulated in patients with ICANS or severe CRS. Box and whisker plot shows all points (vertical bars, min to max points; box, first to third quartile, with median as horizontal bar). # No CRS group included patients with mild phenotype (I-II grade); CRS group included patients with III-IV grade. In green, living patients; in purple, relapsed patient; in black, patients who died.

Article Snippet: The CD19 (130-129-550) and CD22 (130-126-727) CAR detection reagents, anti-CD19-Viogreen (130-113-649), anti-CD3-Viogreen (130-113-142), and anti-biotin-PE (130-110-951) were from Miltenyi Biotec.

Techniques: Expressing, Flow Cytometry, Whisker Assay

Journal: bioRxiv

Article Title: MiniCARbids: Minimalistic human binding domains specifically tailored to CAR T applications

doi: 10.1101/2025.09.09.675083

Figure Lengend Snippet: (A) The K D values of CD22-miniCARbids were determined by titrations of soluble CD22-miniCARbids on NALM6 cells. (B) A representative example of titrations of miniCARbids 22_1611 and 22_1317 on NALM6 cells is shown. The binding intensity was assessed via anti-His-tag staining by flow cytometry. Data were fitted with a 1:1 binding model (solid lines) for the calculation of the respective K D values illustrated in (A) (average ± SD, n=3 or 4, biological replicates). (C) Thermostability of CD22-miniCARbids and their parental protein 5UMR was assessed using DSC (average ± SD of 3 independent measurements, technical replicates). (D) Aggregation properties of CD22-miniCARbids were assessed using SEC-HPLC. One representative analysis (n=3, technical replicates) of CD22-miniCARbids and their parental protein 5UMR is shown. (E) Binding specificity was assessed by incubating NALM6, Raji or Jurkat (CD22-negative) cells with 250 nM CD22-miniCARbid, followed by flow cytometric analysis (one of three biological replicates is shown).

Article Snippet: Selection campaigns started with magnetic bead selections using Dynabeads Biotin Binder (Thermo Fisher Scientific) as described previously., Yeast display selections for miniCARbids against CD22 were based on a soluble, biotinylated CD22 protein (AcroBiosystems, SI2-H82E3).

Techniques: Binding Assay, Staining, Flow Cytometry

Journal: bioRxiv

Article Title: MiniCARbids: Minimalistic human binding domains specifically tailored to CAR T applications

doi: 10.1101/2025.09.09.675083

Figure Lengend Snippet: (A) CAR architecture used for the in vitro assessment of CAR activity. (B) Expression of CARs based on ten CD22-specific miniCARbids and scFvs HA22, m971-1xG 4 S and m971-4xG 4 S as benchmarks in Jurkat Nur77 reporter cells was assessed via anti-MAP-tag staining by flow cytometry (average ± SD, n=3, biological replicates). (C) Activation of CD22-specific CARs in Jurkat Nur77 reporter cells in the presence or absence of a 2-fold excess of NALM6 target cells was assessed via the expression of mKO2 by flow cytometry (average ± SD, n=3, biological replicates). (D) Cytotoxicity of CD22-specific CAR T cells and mock T cells (no CAR) against Raji cells (E:T 2:1, average ± SD, n=4, biological replicates). (E and F) Release of IFN-γ (E) and IL-2 (F) analyzed via ELISA. The cytokines were analyzed in the supernatants of co-cultures with Raji cells (E:T 2:1, average ± SD, n=4, biological replicates). (G) Cytotoxicity of CD22-specific CAR T cells and mock T cells (no CAR) against NALM6 cells (E:T 2:1, average ± SD, n=4, biological replicates). (H and I) Release of IFN-γ (H) and IL-2 (I) analyzed via ELISA. The cytokines were analyzed in the supernatants of co-cultures with NALM6 cells (E:T 2:1, average ± SD, n=4, biological replicates). Statistical analysis was performed using a repeated measure One-Way ANOVA with a Tukey post hoc test (*p < 0.05, **p < 0.01, ***p < 0.001). The statistical analysis for the cytokine concentration was performed using log-transformed values. Parts of this figure were created with BioRender.com.

Article Snippet: Selection campaigns started with magnetic bead selections using Dynabeads Biotin Binder (Thermo Fisher Scientific) as described previously., Yeast display selections for miniCARbids against CD22 were based on a soluble, biotinylated CD22 protein (AcroBiosystems, SI2-H82E3).

Techniques: In Vitro, Activity Assay, Expressing, Staining, Flow Cytometry, Activation Assay, Enzyme-linked Immunosorbent Assay, Concentration Assay, Transformation Assay

Journal: Frontiers in Immunology

Article Title: Unveiling spatial complexity in solid tumor immune microenvironments through multiplexed imaging

doi: 10.3389/fimmu.2024.1383932

Figure Lengend Snippet: Immunophenotyping panel for multiplexed tissue imaging of cancer.

Article Snippet: CD22 , REA340 , 50 , 130-124-223 , FITC , Miltenyi Biotec.

Techniques: Imaging

Journal: Frontiers in Immunology

Article Title: Unveiling spatial complexity in solid tumor immune microenvironments through multiplexed imaging

doi: 10.3389/fimmu.2024.1383932

Figure Lengend Snippet: Deep spatial profiling of human palatine tonsil tissues. (A) Hematoxylin and eosin (H&E) staining after 92 MICS cycles, including the marked epithelium, germinal center (GC), and T cell zone of the lymphoid follicle. MICS DAPI and stroma staining depicting the composition and structure of the tonsil. Markers: Collagen III, collagen IV, fibronectin (all extracellular matrix (ECM), cytokeratin (epithelium), podoplanin (lymphatic vessels), CD105/SM Actin (blood vessels). (B) Immune cell content of a human palatine tonsil comprising T cells (CD3), B cells (CD19/CD20), plasma cells (PCs) (CD38/CD138), NK cells (CD56), granulocytes (CD15/CD66b), mast cells (CD117), macrophages (MΦ) (CD163/CD169/CD206), myeloid dendritic cells (mDCs) (CD11c), and plasmacytoid dendritic cells (pDCs) (CD123). (C) Detailed view on the T cell zone, mainly composed of CD4 + helper T cells (T h ) and CD8 + cytotoxic T cells (T c ), mDCs (CD11c), and PCs (CD38/CD138). (D) Detailed view on the GC-mantle zone border, showing different B cells (CD11b, CD21, CD22), mDCs (CD11c), and PCs (CD38/CD138). (E) Cell annotations of three different tonsil samples plus respective bar graphs of gated cell populations, comparing the cell content between the three tonsil samples. Depicted markers and annotated cell types as indicated by the color code. ROI sizes: 976 x 640 µm, zoomed-in subregions in (C, D) : 334 µm x 219 µm. Scale bar: 100 µm.

Article Snippet: CD22 , REA340 , 50 , 130-124-223 , FITC , Miltenyi Biotec.

Techniques: Staining, Clinical Proteomics

Journal: Arthritis Research & Therapy

Article Title: TWEAK and Fn14 expression in the pathogenesis of joint inflammation and bone erosion in rheumatoid arthritis

doi: 10.1186/ar3294

Figure Lengend Snippet: Dual immunostaining for TWEAK and cell lineage markers and cells expressing Fn14 . Dual immunostaining for TWEAK (red) and CD68 (blue) in inflamed synovial tissue from a patient with active RA ( A ). Dual immunostaining for TWEAK (red) with CD38 (blue) with co-expression of TWEAK and CD38 (purple) indicated by arrow ( B ). C ) and D ) Dual immunostaining for TWEAK (red) with CD22 (blue). E ) TWEAK expression (red) in multinucleated cells (indicated by arrows), and F ) by plasma cells in tonsil tissue. Expression of Fn14 (brown) in multinucleated cells ( G ), and blood vessels of the synovial tissue ( H ), indicated by arrows. Sections shown in E, F, G, and H were counterstained with haematoxylin. Images shown in B and C were obtained with obj ×10; image shown in A obtained with obj ×20, D, F, G, H with obj ×40 and E with obj ×60.

Article Snippet: Anti-TWEAK antibody was combined with MAbs for human cell surface markers: CD68 (macrophage; clone KP-1, Dako), CD22 (B lymphocyte; MAB1968, R&D Systems, Minneapolis, MN, USA), Tryptase G3 (mast cell; Cell Marque, Rocklin, CA, USA) and CD38 (plasma cells, BD Biosciences, Franklin Lakes, NJ, USA).

Techniques: Immunostaining, Expressing, Clinical Proteomics

Journal: Arthritis Research & Therapy

Article Title: TWEAK and Fn14 expression in the pathogenesis of joint inflammation and bone erosion in rheumatoid arthritis

doi: 10.1186/ar3294

Figure Lengend Snippet: TWEAK expression by PBMC . PBMC from two healthy volunteers were sorted by FACS based on their expression of CD22, yielding CD22 + and CD22 - populations of greater than 94% purity based on post-sort analysis ( A ). Isolated cells were then analysed for TWEAK mRNA expression relative to that of GAPDH, by real-time RT-PCR ( B ). Data shown are means of triplicate reactions ± SD. Differences in relative expression of TWEAK mRNA between CD22 + and CD22 - populations were tested by Student's t -test (** P < 0.001).

Article Snippet: Anti-TWEAK antibody was combined with MAbs for human cell surface markers: CD68 (macrophage; clone KP-1, Dako), CD22 (B lymphocyte; MAB1968, R&D Systems, Minneapolis, MN, USA), Tryptase G3 (mast cell; Cell Marque, Rocklin, CA, USA) and CD38 (plasma cells, BD Biosciences, Franklin Lakes, NJ, USA).

Techniques: Expressing, Isolation, Quantitative RT-PCR

Journal: Journal of translational medicine

Article Title: Unraveling resistance mechanisms in anti-CD19 chimeric antigen receptor-T therapy for B-ALL: a novel in vitro model and insights into target antigen dynamics.

doi: 10.1186/s12967-024-05254-z

Figure Lengend Snippet: Fig. 5 Observation of CD19-BBζ-CAR expression in relapsed Nalm-6 cells and salvage treatment. A Detection of FMC63 and CD247 transcripts and 4-1BB gene of CAR in CD19+ Nalm-6 (red) and relapsed CD19− Nalm-6 cells (blue) by qRT-PCR. Data of left bar graph represent the relative quantification using ACTB as the internal reference. Error bars represent s.d. The data are the representative of three independent experiments. B Expression of CD19 and CAR on CD19+ Nalm-6 cells and relapsed CD19− Nalm-6 cells analyzed by flow cytometry (representative of 3 experiments). Merge Graphs, the blue dots represent CD19− Nalm-6 cells and the red dots represent Nalm-6 cells. C Confocal imaging of Nalm-6 cells and relapsed CD19− Nalm-6 cells using Alexa Flour 488-conjugated anti-CD19 antibody (green), Alexa Flour 647-conjugated anti-CAR19 antibody (red), and DAPI (blue). D Lentiviral integration sites of CAR transduced Nalm-6 cells were analyzed by linear-amplification mediated PCR (LAM-PCR) and visualized with Circos plots. The integration sites across the genome and genomic features were shown from outer to inner circle: (1) cytogenetic bands; (2) genes that harbor these integration sites along with a bar chart showing the reads of integration sites; (3) the distribution of integration sites, with colored circles representing different gene functional regions of the host sequence: purple for promoter region, green for intron region, and red for distal intergenic region. E Phenotype changes of Nalm-6 cells transduced with small amount of CD19 CAR lentiviruses detected by flow cytometry over time. Gating was based on the same cells stained with isotype-matched antibody. F Dynamics of CD19− B phenotype in relapsed cells after co-culture with different ratios (5×, 20×) of Nalm-6 cells. Gating was based on the same cells stained with isotype-matched antibody. G Relapsed CD19− Nalm-6 cells were tested by qPCR specific for VSV-G sequence. H Comparison of in vitro efficacy of CD19-, CD22-, CD19/CD22- and CD22×CD19- CAR T cells. Cocultures with the relapsed cells were performed at 1:5, 1:1, and 5:1 E: T ratios, and lysis efficacies were detected by the LDH release assay Declarations

Article Snippet: The cells were then washed twice and stained with phycoerythrin (PE) streptavidin (BD bioscience, USA) for 15 min. CART-22 cells and CART-22/19 cells were washed once and incubated with CD22 Fc Alexa Fluor® 647 Protein (R&D Systems, USA) for 15 min. To detect in vitro cytotoxicity of CART-19 cells, transduced and untransduced T cells were co-cultured with Nalm-6 cells (total 1 × 106 cells) at E: T ratios (0.2:1, 0.5:1, 1:1, 5:1) for 6, 24 and 72 h. Cells were pipetted to incubate with antibodies for 30 min at room temperature in the dark and washed twice with PBS.

Techniques: Expressing, Quantitative RT-PCR, Quantitative Proteomics, Flow Cytometry, Imaging, Amplification, Functional Assay, Sequencing, Transduction, Staining, Co-Culture Assay, Comparison, In Vitro, Lysis, Lactate Dehydrogenase Assay

Journal: bioRxiv

Article Title: Rational redesign of antigen binding domain improves in vivo efficacy of CD22-CAR T cells

doi: 10.1101/2025.03.13.643183

Figure Lengend Snippet: 1A: Schematic: Timeline for in vivo experiment. NSG mice were injected with 1e6 indicated Nalm6 leukemia on day -3, followed by 5e6 CD22-CAR T cells on day 0. Bioluminescent imaging was performed before CAR dosing on day 0, as well on days 5 and 11 post-CAR. 1B: Quantification of bioluminescence data in A. 1C: ELISA measuring Granzyme B in supernatant after 16 hour co-culture of CD22-CAR T cells with the indicated leukemia. 1D: Degranulation as measured by CD107a expression after 4 hour co-culture assay. 1E: Activation as measured by CD69 expression after 6 hour co-culture assay. 1F: Activation as measured by CD25 expression after 24 hour co-culture assay. All in vitro assays performed with n=3 technical replicates, 1 experiment. In vivo assay performed with n=5 mice per group, 1 experiment. Data represent mean +/-SD. * p<0.05, ** p<0.01, *** p<0.001, **** p<0.0001.

Article Snippet: Following expansion, transduction efficiency of the CD22-CAR was evaluated by flow cytometry staining with CD22-Protein Fc (R&D Systems) and CAR T cells were cryopreserved or used immediately for in vitro or in vivo assays.

Techniques: In Vivo, Injection, Imaging, Enzyme-linked Immunosorbent Assay, Co-Culture Assay, Expressing, Co-culture Assay, Activation Assay, In Vitro

Journal: bioRxiv

Article Title: Rational redesign of antigen binding domain improves in vivo efficacy of CD22-CAR T cells

doi: 10.1101/2025.03.13.643183

Figure Lengend Snippet: 2A: Flow cytometry plots showing IL-2 by IFNg production after 6 hour coculture of the indicated CD22-CAR T cell with the indicated leukemia. 2B: Quantification of cytokine data in A. 2C: Schematic: Timeline for in vivo experiment. NSG mice were injected with 1e6 indicated Nalm6 leukemia on day -3, followed by 4e6 CD22-CAR T cells on day 0. Bioluminescent imaging was performed before CAR dosing on day -1, and biweekly post-CAR injection. Mice were monitored for survival. 2D: Quantification of bioluminescence data against WT leukemia from C. 2E: Survival of mice bearing WT leukemia. 2D: Quantification of bioluminescence data against CD22 Lo leukemia from C. 2E: Survival of mice bearing CD22 Lo leukemia. All in vitro assays performed with n=3 technical replicates, and are representative of two experiments with two independent donors. In vivo assay performed with n=5 mice per group, 1 experiment. Data represent mean +/-SD. * p<0.05, ** p<0.01, *** p<0.001, **** p<0.0001.

Article Snippet: Following expansion, transduction efficiency of the CD22-CAR was evaluated by flow cytometry staining with CD22-Protein Fc (R&D Systems) and CAR T cells were cryopreserved or used immediately for in vitro or in vivo assays.

Techniques: Flow Cytometry, In Vivo, Injection, Imaging, In Vitro

Journal: bioRxiv

Article Title: Rational redesign of antigen binding domain improves in vivo efficacy of CD22-CAR T cells

doi: 10.1101/2025.03.13.643183

Figure Lengend Snippet: S1A: Cell-based direct antigen-binding affinity titration assay. Indicated CARs were stained with indicated concentrations of fluorophore-conjugated CD22 Protein Fc. MFI of CAR+ Populations were measured and normalized to peak protein binding for each individual CAR. Data represents one experiment with one replicate per concentration.

Article Snippet: Following expansion, transduction efficiency of the CD22-CAR was evaluated by flow cytometry staining with CD22-Protein Fc (R&D Systems) and CAR T cells were cryopreserved or used immediately for in vitro or in vivo assays.

Techniques: Binding Assay, Titration, Staining, Protein Binding, Concentration Assay

Journal: bioRxiv

Article Title: Rational redesign of antigen binding domain improves in vivo efficacy of CD22-CAR T cells

doi: 10.1101/2025.03.13.643183

Figure Lengend Snippet: Figures S2A to S2F quantify indicated metrics by flow cytometry after coculture of indicated CD22-CAR with indicated leukemia after 6 hour coculture. S2A: %+ and MFI for IFNg production against WT leukemia. S2B: %+ and MFI for IL2 production against WT leukemia. S2C: %+ of cells making IFNg and IL-2 against WT leukemia. S2D: %+ and MFI for IFNg production against CD22 Lo leukemia. S2E: %+ and MFI for IL2 production against CD22 Lo leukemia. S2F: %+ of cells making IFNg and IL-2 against CD22 Lo leukemia. Figures S2G to S2J quantify CAR and leukemia counts relative to a starting 5:1 ratio of leukemia and CAR to fluorescent counting beads. Aliquots were taken from each condition and analyzed by flow cytometry at each of the indicated time points. S2G: Quantification of CAR Count/Bead Count ratio against WT leukemia. S2H: Quantification of Leukemia Count/Bead Count ratio for WT leukemia. S2I: Quantification of CAR Count/Bead Count ratio against CD22 Lo leukemia. S2J: Quantification of Leukemia Count/Bead Count ratio for CD22 Lo leukemia. All in vitro assays performed with n=3 technical replicates. are representative of two experiments with two independent donors. are representative of one experiment. Data represent mean +/- SD. * p<0.05, ** p<0.01, *** p<0.001, **** p<0.0001.

Article Snippet: Following expansion, transduction efficiency of the CD22-CAR was evaluated by flow cytometry staining with CD22-Protein Fc (R&D Systems) and CAR T cells were cryopreserved or used immediately for in vitro or in vivo assays.

Techniques: Flow Cytometry, In Vitro

Journal: bioRxiv

Article Title: Rational redesign of antigen binding domain improves in vivo efficacy of CD22-CAR T cells

doi: 10.1101/2025.03.13.643183

Figure Lengend Snippet: 3A: Schematic: Timeline for in vivo experiment. NSG mice were injected with 1e6 WT Nalm6 leukemia on day-3, followed by 2e6 of indicated CD22-CAR T cells on day 0. Bioluminescent imaging was performed before CAR dosing on day -1, and biweekly post-CAR. 3B: Quantification of average bioluminescence data for each group in A. 3C: Quantification of individual bioluminescence data for each group in A. 3D: Schematic: Timeline for in vivo experiment. NSG mice were injected with 1e6 WT Nalm6 leukemia on day -3, followed by 4e6 of indicated CD22-CAR T cells on day 0. Bioluminescent imaging was performed before CAR dosing on day -1, and biweekly post-CAR. 3E: Quantification of average bioluminescence data for each group in D. 3F: Survival of mice treated with 4e6 of the indicated CAR T cells. In vivo assay performed with n=5 mice per group, 1 experiment (3A to 3C) or 3 experiments with independent donors (3D to 3F). 3D to 3E are representative data from one experiment. 3F is pooled data, SA-SL (n=15), HA-SL (n=15), HA-LL (n=15). Data represent mean +/- SD. * p<0.05, ** p<0.01, *** p<0.001, **** p<0.0001.

Article Snippet: Following expansion, transduction efficiency of the CD22-CAR was evaluated by flow cytometry staining with CD22-Protein Fc (R&D Systems) and CAR T cells were cryopreserved or used immediately for in vitro or in vivo assays.

Techniques: In Vivo, Injection, Imaging

Journal: bioRxiv

Article Title: Rational redesign of antigen binding domain improves in vivo efficacy of CD22-CAR T cells

doi: 10.1101/2025.03.13.643183

Figure Lengend Snippet: 4A: Schematic: Timeline for in vivo experiment. NSG mice were injected with 1e6 CD22 Lo Nalm6 leukemia on day -3, followed by 4e6 of indicated CD22-CAR T cells on day 0. Bioluminescent imaging was performed before CAR dosing on day -1, and biweekly post-CAR. 4B: Quantification of average bioluminescence data for each group in A. 5C: Quantification of individual bioluminescence data for each group in A. For 4D to 4E, bone marrow was analyzed by flow cytometry at day 18 post-CAR for indicated cell population. 4D: % CAR+ of live marrow. 4E: % leukemia of live marrow. 4F: Survival of mice treated with 4e6 of indicated CAR T cells. In vivo assay performed with n=5 mice per group, 4 experiments with independent donors. Data in 4A to 4C is representative data from one experiment. Survival is pooled from 3 experiments with independent donors: Mock (n=10), SA-SL (n=15), HA-SL (n=10), HA-LL (n=15). Data represent mean +/- SD. * p<0.05, ** p<0.01, *** p<0.001, ****

Article Snippet: Following expansion, transduction efficiency of the CD22-CAR was evaluated by flow cytometry staining with CD22-Protein Fc (R&D Systems) and CAR T cells were cryopreserved or used immediately for in vitro or in vivo assays.

Techniques: In Vivo, Injection, Imaging, Flow Cytometry