Journal: Scientific Reports

Article Title: Positive and negative cooperativity of TNF and Interferon-γ in regulating synovial fibroblast function and B cell survival in fibroblast/B cell co-cultures

doi: 10.1038/s41598-020-57772-7

Figure Lengend Snippet: Antibodies used for flow cytometry.

Article Snippet: Miltenyi , 130-097-656 , CD268 (BAFF-R) , FITC.

Techniques: Cytometry, Membrane

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

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: In vitro and in vivo activation induces BAFF and APRIL expression in B cells.

doi: 10.4049/jimmunol.179.9.5947

Figure Lengend Snippet: FIGURE 3. In vitro and in vivo activation induces BAFF and APRIL expression. Sorted splenic B cells were in vitro activated with medium alone, LPS, or CpG-ODN for 3 days. Cytospins were prepared and costained with anti- (green) and polyclonal rabbit anti-BAFF (red; A) or anti-APRIL Ab (red; B). Nuclei were counterstained with 4,6-dia- midino-2-phenylindole (DAPI; blue). Single staining and overlays are shown. C, Splenic tissue sections from immunized (C1–3), naive BALB/c (C4), and 3- to 4-mo-old (NZB NZW)F1 mice (D) are shown. Consecutive sections (C1,2) were double stained with anti-BAFF and either anti-FDC (M2) or PNA. Light zone (LZ) and dark zone (DZ) are indicated. E, Splenic tissue sec- tions from 6- to 7-mo-old (NZB NZW)F1 mice were stained for , BAFF, and APRIL. Single staining and overlays of plasma cells are shown. Original magnification, 40. Representative results from three in- dependent experiments are shown.

Article Snippet: To analyze BAFF expression in GC, tissue sections were double stained with polyclonal rabbit anti-BAFF and biotinylated FDC-specific mAb M2 (Serotec) or biotinylated PNA (Vector Laboratories).

Techniques: In Vitro, In Vivo, Activation Assay, Expressing, Staining, Clinical Proteomics