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. 2021 Jul 23;12:652223. doi: 10.3389/fimmu.2021.652223

Figure 1.

Figure 1

Generation of S309-CAR-NK-92MI cells. (A) Plasmid construct of S309-CAR. The SFG retroviral vector contains the S309 single chain antibody fragment (accession code 6WS6 on PDB), a human IgG1 CH2CH3 hinge region, a CD28 transmembrane region, followed by the co-stimulatory CD28, 4-1BB, and the intracellular domain of CD3ζ. (B) Determination of S309-CAR-NK expression by flow cytometry. S309-CAR cells were collected and stained with anti-CD56 and CAR F(ab)2 domain [IgG (H+L)] for flow cytometry. The cells were then sorted to achieve a homogenous population of high CAR expression. (C) Representative histogram showing the immunoprofiling of S309-CAR-NK-92MI cells by flow cytometry. S309-CAR-NK-92MI or the wildtype (WT) NK-92MI cells were stained with antibodies against different immunomodulatory receptors including CTLA-4, PD-1, NKG2A, NKG2C, NKG2D, NKp46, CD56, CD16, 2B4, DNAM-1, CD94, KLRG1, TIM-3, LAG-3, and TIGIT.