Journal: Antibodies

Article Title: Evaluating the Therapeutic Efficacy of an Anti-BAFF Receptor Antibody Using a Rheumatoid Arthritis Mouse Model

doi: 10.3390/antib14040090

Figure Lengend Snippet: Antibody-mediated dose-dependent inhibition of B220+/CD21+CD23+ B-cell proliferation analyzed by flow cytometry. ( A ) CD21+ CD23+ cells, ( B ) B cells. Splenocytes from C57BL/6 mice (1 × 10 6 cells/mL) were treated with or without the following: BAFF [7 ng/mL]. mIgG2a isotype control mAb (clone-c1.18.4), mIgG1 isotype control mAb (clone-MOPC-21), V3-1 mIgG2a mAb, V3-46s mIgG2a mAb, and V3-46s mIgG1 mAb were added in a dose-dependent manner, as indicated, with 3-fold dilutions starting at 3 µg/mL, and cultured for 72 h at 37 °C. Cells were then stained for B220, CD21, and CD23, and visualized on a CytoflexL4.

Article Snippet: Cells were then stained for CD45 (Biolegend, catalog BLG-103108), B220 (Miltenyi, catalog 130-110-846), CD21/CD35 (Miltenyi, catalog 130-111-732), CD23 (Miltenyi, catalog 130-118-761), and DAPI (Sigma, catalog D9542), and were visualized on a CytoflexL4 (Beckman Coulter) flow cytometer.

Techniques: Inhibition, Flow Cytometry, Control, Cell Culture, Staining

Journal: Blood

Article Title: Viral latency locus augments B-cell response in vivo to induce chronic marginal zone enlargement, plasma cell hyperplasia, and lymphoma.

doi: 10.1182/blood-2012-03-415620

Figure Lengend Snippet: Figure 3. MZ expansion in KSHV latency locus transgenic mice. (A) Splenocytes from wild-type (WT) or latency transgenic mouse (TG) were analyzed by fluorescence- activated cell sorter (FACS). CD191CD241 cells were further gated with CD21 and CD23. Percentages of CD191CD241CD21hiCD23- cells among whole lymphocytes are indicated in FACS profiles. (B) Alternatively, CD191IgD- cells were gated with CD21 and CD23 in an independent analysis. (C) Bar plot of flow cytometry data from panels A-B (n 5 5 for WT and n 5 9 for the transgenic mice). (D) MZ precursor cells were gated as CD191IgD1CD21hiCD231. Average percentages of 6 WT and 9 latency transgenic (TG) mice are shown with the standard deviation. (E) Frozen spleen sections were stained with B220 (green) for B cells and MOMA-1 for marginal zone macrophages (red). Magnification 3200. FL, follicle.

Article Snippet: Antibodies and reagents used for flow cytometry and immunohistology The following antibodies were used: anti-vCYC, biotinylated MOMA-1 (Abcam), rat mAb to LANA (Advanced Biotechnologies Inc.), Alexa Fluor 488-conjugated anti-B220, fluorescein isothiocyanate-conjugated anti-mouse activation antigen GL7, anti-mouse IgD, phycoerythrin (PE)-conjugated anti-mouse CD21/CD35, anti-mouse IgM, PE/Cy5.5-conjugated anti-mouse CD45R, PE-conjugated anti-mouse CD138 (BD Biosciences), Alexa Fluor 555-conjugated anti-rat IgG antibodies (Cell Signaling Technology), Alexa Fluor 488-conjugated streptavidin, Alexa Fluor 555-conjugated streptavidin, allophycocyanin-conjugated anti-mouse CD19, anti-mouse CD23, fluorescein isothiocyanate-conjugated anti-mouse CD24, anti-mouse CD45R (B220), anti-mouse CD69, and PE-conjugated anti-mouse CD86 (Invitrogen), antikaposin (Millipore), PE-conjugated mouse anti-major histocompatibility complex class II (Miltenyi), and biotinylated PNA (Vector Laboratories).

Techniques: Transgenic Assay, Fluorescence, Flow Cytometry, Standard Deviation, Staining