Figure 2.

CD34-hinged CAR expression and enrichment in primary human NK cells. Primary human NK cells were transduced with the lentiviral vectors delineated in panels (A, B) encoding BFP in cis with CD34-hinged CD33, CD123, CD19, or Cetux CARs. (B) Schematic representation of CAR construct including CD34-derived C6 hinge for microbead interactions. (C) Three to four days after lentiviral CAR transduction, NK cells were analyzed for BFP and CAR (via CD34-PE staining) expression by flow cytometry. Representative dot plots are shown. (D, E) CD34-hinged CAR NK cells were enriched via magnetic cell sorting using CD34 microbeads and three fractions (before MACS, flowthrough, and after MACS) were analyzed for the content of NK cells expressing BFP by flow cytometry. The histogram blots show representative data. Data are represented as mean ± SEM of at least four biological replicates.