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Fig. 1. B cells populations, BAFF receptors expression, In-vitro B cells specific cytotoxicity, and the cytokine release of BAFF CAR-T cells. A) Percentage of B cell subset populations in healthy (N = 3) vs SLE patient samples (N = 6). The average percentages from control and SLE patients are plotted as pie chart, and the gating strategies are given in the Supplementary Fig. S2A. B) The BAFF receptors expression in the healthy donor (N = 8) and active SLE patient peripheral blood monocytic cells (PBMCs) gated on the lymphocytes (N = 13). C) Percentage of the BAFF CAR positive T cells after transfection in healthy vs SLE patients (N = 3). D) In-vitro cytotoxicity of autologous control-T and BAFF CAR-T cells on healthy donor B cells and, E) SLE patient B cells, co-cultured at the E:T ratio of 1:1, 3:1, and 5:1 for 48 h (N = 3). Also, the effect of control-T and BAFF CAR-T cells from healthy donors and SLE on Jeko-1, RPMI-8226, and Jeko-KO B cell lines at 3:1 ratio (N = 3). F) Representative 2D scatter plots showing the CD19 vs <t>BAFFr/BCMA/TACI/IgM</t> populations after co- culturing the UT/CD19 CAR-T/BAFF-CAR-T cells with the active SLE patient PBMCs at E:T ratio of 5:1 for 24 h gated on lymphocytes. G) Percentage of CD19+ BAFFr+, CD19+ IgM+, and CD19−IgM+, and the CD19−BCMA+ populations after co-culture (N = 5). H) Inflammatory cytokines, activation/degranulation enzyme levels in the supernatant after co-culturing the PBMCs with UT/CD19-CAR-T/BAFF CAR-T cells for 24 h at E:T ratio of 5:1 (N = 4). I) In-vitro cytotoxicity of UT, CD19 CAR-T and BAFF CAR-T cells on CD138+ plasma B cells from multiple myeloma patient samples after co-culture for 24 h at E:T ratios of 5:1 and 10:1 (N = 3). The p value < 0.05 is considered as statistically significant. The student t-test was employed to compare figure B, C, G, and two-way ANOVA with Tukey’s test was used for the analysis of the figures A, D, E, H and I. P < 0.05 = *, P < 0.01 = **, P < 0.001 = ***.
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Fig. 1. B cells populations, BAFF receptors expression, In-vitro B cells specific cytotoxicity, and the cytokine release of BAFF CAR-T cells. A) Percentage of B cell subset populations in healthy (N = 3) vs SLE patient samples (N = 6). The average percentages from control and SLE patients are plotted as pie chart, and the gating strategies are given in the Supplementary Fig. S2A. B) The BAFF receptors expression in the healthy donor (N = 8) and active SLE patient peripheral blood monocytic cells (PBMCs) gated on the lymphocytes (N = 13). C) Percentage of the BAFF CAR positive T cells after transfection in healthy vs SLE patients (N = 3). D) In-vitro cytotoxicity of autologous control-T and BAFF CAR-T cells on healthy donor B cells and, E) SLE patient B cells, co-cultured at the E:T ratio of 1:1, 3:1, and 5:1 for 48 h (N = 3). Also, the effect of control-T and BAFF CAR-T cells from healthy donors and SLE on Jeko-1, RPMI-8226, and Jeko-KO B cell lines at 3:1 ratio (N = 3). F) Representative 2D scatter plots showing the CD19 vs BAFFr/BCMA/TACI/IgM populations after co- culturing the UT/CD19 CAR-T/BAFF-CAR-T cells with the active SLE patient PBMCs at E:T ratio of 5:1 for 24 h gated on lymphocytes. G) Percentage of CD19+ BAFFr+, CD19+ IgM+, and CD19−IgM+, and the CD19−BCMA+ populations after co-culture (N = 5). H) Inflammatory cytokines, activation/degranulation enzyme levels in the supernatant after co-culturing the PBMCs with UT/CD19-CAR-T/BAFF CAR-T cells for 24 h at E:T ratio of 5:1 (N = 4). I) In-vitro cytotoxicity of UT, CD19 CAR-T and BAFF CAR-T cells on CD138+ plasma B cells from multiple myeloma patient samples after co-culture for 24 h at E:T ratios of 5:1 and 10:1 (N = 3). The p value < 0.05 is considered as statistically significant. The student t-test was employed to compare figure B, C, G, and two-way ANOVA with Tukey’s test was used for the analysis of the figures A, D, E, H and I. P < 0.05 = *, P < 0.01 = **, P < 0.001 = ***.

Journal: Journal of autoimmunity

Article Title: CAR-T cell targeting three receptors on autoreactive B cells for systemic lupus erythematosus therapy.

doi: 10.1016/j.jaut.2025.103369

Figure Lengend Snippet: Fig. 1. B cells populations, BAFF receptors expression, In-vitro B cells specific cytotoxicity, and the cytokine release of BAFF CAR-T cells. A) Percentage of B cell subset populations in healthy (N = 3) vs SLE patient samples (N = 6). The average percentages from control and SLE patients are plotted as pie chart, and the gating strategies are given in the Supplementary Fig. S2A. B) The BAFF receptors expression in the healthy donor (N = 8) and active SLE patient peripheral blood monocytic cells (PBMCs) gated on the lymphocytes (N = 13). C) Percentage of the BAFF CAR positive T cells after transfection in healthy vs SLE patients (N = 3). D) In-vitro cytotoxicity of autologous control-T and BAFF CAR-T cells on healthy donor B cells and, E) SLE patient B cells, co-cultured at the E:T ratio of 1:1, 3:1, and 5:1 for 48 h (N = 3). Also, the effect of control-T and BAFF CAR-T cells from healthy donors and SLE on Jeko-1, RPMI-8226, and Jeko-KO B cell lines at 3:1 ratio (N = 3). F) Representative 2D scatter plots showing the CD19 vs BAFFr/BCMA/TACI/IgM populations after co- culturing the UT/CD19 CAR-T/BAFF-CAR-T cells with the active SLE patient PBMCs at E:T ratio of 5:1 for 24 h gated on lymphocytes. G) Percentage of CD19+ BAFFr+, CD19+ IgM+, and CD19−IgM+, and the CD19−BCMA+ populations after co-culture (N = 5). H) Inflammatory cytokines, activation/degranulation enzyme levels in the supernatant after co-culturing the PBMCs with UT/CD19-CAR-T/BAFF CAR-T cells for 24 h at E:T ratio of 5:1 (N = 4). I) In-vitro cytotoxicity of UT, CD19 CAR-T and BAFF CAR-T cells on CD138+ plasma B cells from multiple myeloma patient samples after co-culture for 24 h at E:T ratios of 5:1 and 10:1 (N = 3). The p value < 0.05 is considered as statistically significant. The student t-test was employed to compare figure B, C, G, and two-way ANOVA with Tukey’s test was used for the analysis of the figures A, D, E, H and I. P < 0.05 = *, P < 0.01 = **, P < 0.001 = ***.

Article Snippet: The K562 luciferase cells were purchased from ATCC (CAT: CCL-243Luc2, Lot: 70014924), and transduced with lentivirus particles encoding BAFFr (Origene technologies, CAT No: RC211270L4V), or NgR (G&P Biosciences, CAT No: LTV9002P), constructs encoding puromycin resistance gene.

Techniques: Expressing, In Vitro, Control, Transfection, Cell Culture, Co-Culture Assay, Activation Assay, Clinical Proteomics

The effect of CCN1 on gene expression of APRIL/BAFF system (fold change).

Journal: Scientific Reports

Article Title: CCN1 promotes APRIL/BAFF signaling in esophageal squamous cell carcinoma but attenuates it in esophageal adenocarcinoma

doi: 10.1038/s41598-025-86228-z

Figure Lengend Snippet: The effect of CCN1 on gene expression of APRIL/BAFF system (fold change).

Article Snippet: The following primary antibodies were used in this study: CCN1 (TA349858) and BCMA (TA382649) from Origene, APRIL (sc-374674), BAFF (sc-271809), TACI (sc-365253), BAFFR (sc-365409), Furin (sc-133142), and β-actin (sc69879) from Santa Cruz Biotechnology.

Techniques: Gene Expression