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. 2023 Jan 7;12(1):2163784. doi: 10.1080/2162402X.2022.2163784

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Figure 7. — Pomalidomide increases CD38 expression of PEL cell lines and enhances Dara-mediated ADCC. (a) PEL cell lines were treated for 72 h with DMSO control (0 Pom) or indicated concentrations of Pom and then the live cell number was counted after trypan blue-staining. Live cell number in Pom-treated cells as a percentage of DMSO-treated cells are presented. (b) Levels of surface CD38 on PEL cell lines treated with DMSO or Pom for 72 h were measured by flow cytometry using PerCP-Cy5.5 conjugated anti-CD38 antibody and expressed as median fluorescence intensity (MFI) after subtracting background MFI from isotype control ab. Results represent average MFIs and standard deviations from at least 3 separate experiments. Because of toxicity at higher concentrations, the highest concentration of Pom utilized in JSC-1 cells was 1 µM. (c) ADCC pathway activation by DMSO or Pom-treated PEL cells (BC-2 with 1 µM and JSC-1 with 0.5 µM Pom) after co-incubation with Jurkat-ADCC cells for 6 h in the presence of various concentrations of Dara. Data show one representative experiment from three separate experiments and is expressed as relative luminescence unit (RLU) from Jurkat-ADCC cells alone or co-incubated with the PEL lines. (d) Change in ADCC-induction by Pom-treated BC-2 and JSC-1 cells over that by DMSO-treated cells is presented as fold change in RLU from Jurkat-ADCC cells after 6 h co-incubation in the presence of 0.6 µg/mL Dara or isotype control ab. Data show averages and standard deviations from 4 separate experiments. Statistically significant differences (*P ≤ .05, **P ≤ .01) by 2-tailed t-test are indicated.