pd-l1 t-hank  (ImmunityBio Inc)

Journal: Cancer research communications

Article Title: Combinatorial Natural Killer Cell–based Immunotherapy Approaches Selectively Target Chordoma Cancer Stem Cells

doi: 10.1158/2767-9764.crc-21-0020

Figure Lengend Snippet: Six chordoma cell lines; established from 3 from clival tumor patients (UM-Chor1, MUG-CC1, UM-Chor5), and 3 sacral tumor patients (JHC7, U-CH1, U-CH2) were analyzed by flow cytometry to quantify expression surface markers MHC-I/HLA-A,B,C, PD-L1, and EGFR. Cell surface expression of each marker is reported in % positive cells and MFI.

Article Snippet: Immune reagents and NK cells An anti-PD-L1 antibody (N-601), an IL-15/IL-15r superagonist (N-803), and the PD-L1-specific chimeric antigen receptor engineered high affinity natural killer cells (PD-L1 t-haNKs) were provided by ImmunityBio under a Cooperative Research and Development Agreement (CRADA) with the National Cancer Institute (NCI) of the National Institutes of Health (NIH).

Techniques: Flow Cytometry, Expressing, Marker

Journal: Cancer research communications

Article Title: Combinatorial Natural Killer Cell–based Immunotherapy Approaches Selectively Target Chordoma Cancer Stem Cells

doi: 10.1158/2767-9764.crc-21-0020

Figure Lengend Snippet: Two chordoma cell lines (UM-Chor1, JHC7) were used as targets for PD-L1 t-haNK cells in 111In-release killing assays. (A) Chordoma cells were analyzed by flow cytometry for PD-L1 expression after 24-hour co-incubation with IFNγ on the same day as killing assays. Cell surface expression of PD-L1 quantified by flow cytometry is reported in % positive cells and MFI. Bold values denote an increase of > 30% relative to untreated control cells. (B) Chordoma cells were co-incubated with IFNγ or left untreated as controls for 24 hours before being used as targets for PD-L1 t-haNK cells. All E:T ratios are 20:1. Statistical analyses were done by one-way ANOVA with Tukey’s multiple comparisons test. *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, ****P ≤ 0.0001. Results shown are the means ± SEM of technical triplicate measurements and are representative of three independent experiments.

Article Snippet: Immune reagents and NK cells An anti-PD-L1 antibody (N-601), an IL-15/IL-15r superagonist (N-803), and the PD-L1-specific chimeric antigen receptor engineered high affinity natural killer cells (PD-L1 t-haNKs) were provided by ImmunityBio under a Cooperative Research and Development Agreement (CRADA) with the National Cancer Institute (NCI) of the National Institutes of Health (NIH).

Techniques: Flow Cytometry, Expressing, Incubation, Control

Journal: Cancer research communications

Article Title: Combinatorial Natural Killer Cell–based Immunotherapy Approaches Selectively Target Chordoma Cancer Stem Cells

doi: 10.1158/2767-9764.crc-21-0020

Figure Lengend Snippet: CellTrace Violet-stained UM-Chor1 cells were used as targets for healthy donor NK cells in a flow cytometry-based killing assay. (A) CSC and non-CSC subpopulations were identified using the gating strategy described. (B) UM-Chor1 cells were co-incubated with N-601 or isotype control antibody and NK cells were treated with N-803 where indicated. CSC and non-CSC cell death through N-601-mediated ADCC by untreated or N-803-treated NK cells were evaluated via flow cytometry using a live/dead fixable cell stain exclusion method. (Cand D) Flow cytometry was used to quantify expression of HLA-A,B,C, MICA/B, B7-H6, and PD-L1 expression on CSC and non-CSC UM-Chor1 cells. Cell surface expression of each marker is reported in % positive cells and MFI. Bold values denote an increase of > 20% when comparing CSC vs. non-CSC. All E:T ratios are 5:1. Statistical analyses were done by one-way ANOVA with Tukey’s multiple comparisons test. *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, ****P ≤ 0.0001. Results shown are the means ± SEM of technical quadruplicate measurements and are representative of two independent experiments.

Article Snippet: Immune reagents and NK cells An anti-PD-L1 antibody (N-601), an IL-15/IL-15r superagonist (N-803), and the PD-L1-specific chimeric antigen receptor engineered high affinity natural killer cells (PD-L1 t-haNKs) were provided by ImmunityBio under a Cooperative Research and Development Agreement (CRADA) with the National Cancer Institute (NCI) of the National Institutes of Health (NIH).

Techniques: Staining, Flow Cytometry, Incubation, Control, Expressing, Marker