“Adherent” versus Other Isolation Strategies for Expanding Purified, Potent, and Activated Human NK Cells for Cancer Immunotherapy

Figure 1

Select (low, medium, and high percent expanded NK cells; Refer to bold cases in Tables 1(a) and 1(b)) flow cytometry dot blots of phenotype of (a) cancer patient and (b) healthy donor PBMCs and corresponding expanded cells, and cytotoxicity of expanded cells against K-562 cells. PBMC populations and two-week expanded cells were stained by incubating with Pacific Blue conjugated anti-CD3 (Clone HIT3a, Biolegend) and Phycoerythrin (PE/Cy7) conjugated anti-CD56 antibodies (Clone MEM-188, Biolegend) and fixed in 2% paraformaldehyde. For all flow cytometry experiments, appropriate IgG isotype controls were used to assess nonspecific staining. Cells were analyzed using a BD LSRII FACS flow cytometer and the data was processed using FlowJo Flow Cytometry Analysis Software (TreeStar Inc.). Cytotoxicity of expanded cells against CFSE-labeled K-562 cells was carried out at various E : T using 7-AAD/CFSE cell-mediated cytotoxicity assay kit (Abnoa).

(a)

(b)