Journal: EJHaem

Article Title: Ars2‐containing bispecific, Fab‐ and IgG1‐format BAR‐bodies to target DLBCL cells

doi: 10.1002/jha2.635

Figure Lengend Snippet: Characterization of bispecific Ars2 BAR‐bodies. Flow cytometric assessment of both bispecific Ars2 BAR‐bodies are shown in parts (A) and (B). Anti‐CD16/Ars2 and anti‐CD3/Ars2 both show no binding to cells of the mantle cell lymphoma (MCL) cell line MAVER‐1 expressing LRPAP1‐reactive B‐cell receptors (BCRs) (A). The LRPAP1/ETA′ immunotoxin served as positive control. Anti‐CD16/Ars2 and anti‐CD3/Ars2 show strong binding to U2932 cells expressing Ars2‐reactive BCRs, whereas LRPAP1/ETA′ does not bind to U2932 cells (B). When incubated with peripheral blood mononuclear cells (PBMCs) as effector cells, both bispecific BAR‐bodies had no effect on MAVER‐1 cells at any concentration (C) but induced lysis of U2932 cells in a concentration‐dependent manner (D). All experiments were performed in triplicate (error bars).

Article Snippet: OCI‐Ly3 cells and the mantle cell lymphoma (MCL) cell lines MAVER‐1 and Granta‐519 were purchased from DSMZ (Braunschweig, Germany).

Techniques: Binding Assay, Expressing, Positive Control, Incubation, Concentration Assay, Lysis

Journal: EJHaem

Article Title: Ars2‐containing bispecific, Fab‐ and IgG1‐format BAR‐bodies to target DLBCL cells

doi: 10.1002/jha2.635

Figure Lengend Snippet: IgG‐format BAR bodies incorporating Ars2, LRPAP1, and Neurabin‐I were tested for binding to the diffuse large B‐cell lymphoma (DLBCL) cell lines U2932, OCI‐Ly3 (Ars2‐reactive B‐cell receptor [BCR]), and HBL‐1 (no Ars2‐reactive BCR) as well as to the mantle cell lymphoma (MCL) cell lines Granta‐519 (BCR of unknown reactivity) and MAVER‐1 (LRPAP1‐reactive BCR). Ars2 BAR‐bodies showed binding to OCI‐Ly3 and U2932 cells but not to other cell lines. LRPAP1 BAR‐bodies served as positive control and showed binding to MAVER‐1 cells as they express BCRs with known LRPAP1 reactivity. Neurabin‐I BAR‐bodies were used as negative controls and showed no binding to any used cell line.

Article Snippet: OCI‐Ly3 cells and the mantle cell lymphoma (MCL) cell lines MAVER‐1 and Granta‐519 were purchased from DSMZ (Braunschweig, Germany).

Techniques: Binding Assay, Positive Control

Journal: EJHaem

Article Title: Ars2‐containing bispecific, Fab‐ and IgG1‐format BAR‐bodies to target DLBCL cells

doi: 10.1002/jha2.635

Figure Lengend Snippet: Cytotoxicity assays (LDH‐release) of IgG‐format BAR‐bodies incorporating Ars2 and LRPAP1 against diffuse large B‐cell lymphoma (DLBCL) cell lines U2932, OCI‐Ly3, and HBL‐1 as well as mantle cell lymphoma (MCL) cell lines Granta‐519 and MAVER‐1. LDH‐release assays with Ars2 BAR‐bodies on HBL‐1 and U2932 cells were performed with and without effector cells (peripheral blood mononuclear cells [PBMCs]). Both Ars2 and LRPAP1 incorporating IgG‐format BAR‐bodies had no effect on HBL‐1 cells without (A) or with (B) effector cells. Ars2, but not LRPAP1 BAR‐bodies, induced specific lysis in U2932 cells. Ars2 BAR‐bodies showed cytotoxic effects (specific lysis of approximately 20% at 10 μg/ml) on U2932 cells even without effector cells (C). This effect increases to a specific lysis of approximately 60% after the addition of PBMCs (D). Similar results could be obtained with OCI‐Ly3 cells as targets, which also express B‐cell receptors (BCRs) with Ars2‐reactivity (E). LRPAP1 BAR‐bodies had no effect on either U2932 or OCI‐Ly3 cells. The MCL cell line Granta‐519 was used as additional negative control (F). LRPAP1 BAR‐bodies and the LRPAP1‐reactive MCL cell line MAVER‐1 were used as positive control (G). All experiments were performed in triplicate (error bars).

Article Snippet: OCI‐Ly3 cells and the mantle cell lymphoma (MCL) cell lines MAVER‐1 and Granta‐519 were purchased from DSMZ (Braunschweig, Germany).

Techniques: Lysis, Negative Control, Positive Control