Journal: Hepatology Communications

Article Title: Human V H -based chimeric antigen receptor T cells targeting glypican 3 eliminate tumors in preclinical models of HCC

doi: 10.1097/HC9.0000000000000022

Figure Lengend Snippet: Engineered CAR T cells display increased cytoxicity in a panel of HCC line. (A) Map of CARs targeting GPC3 with varying hinge and transmembrane domains. (B) CAR constructs resulting in different permutations of CAR constructs. (C) Jurkat-CAR T cells binding to cetuximab, GPC3 at 1 μg/mL. (D) Overview of production of CAR T cells starting from donor PBMCs. (E) Cell count monitored over the course of 11 days. Transduction efficiency of all constructs used in the cytotoxicity experiment. Cell killing graphs showing cytotoxicity. CAR T cells were cultured with GPC3 expressing HCC cells lines (Hep3B, HepG2, and Huh7) and GPC3 knockout (KO) cells at different effector to target ratios. CAR, chimeric antigen receptor; PBMC, peripheral blood mononuclear cell.

Article Snippet: The cell surface binding the CAR on Jurkat cells to GPC3 protein tagged with hfc (ACRO Biosystems) was assessed using anti-human IgG-phycoerythrin (APC)-conjugated antibody (Jackson ImmunoResearch).

Techniques: Construct, Binding Assay, Cell Counting, Transduction, Cell Culture, Expressing, Knock-Out

Journal: Hepatology Communications

Article Title: Human V H -based chimeric antigen receptor T cells targeting glypican 3 eliminate tumors in preclinical models of HCC

doi: 10.1097/HC9.0000000000000022

Figure Lengend Snippet: Fc containing CAR T cells do not improve efficacy of engineered CAR T cells in vitro or in vivo . (A) CAR T-cell construct map with Fc containing long (IgG4-CH2CH3H), intermediate (IgG4-CH3H), and short (IgG4H) hinge regions. (B) Cell count monitored over the course of 14 days. Transduction efficiency measured by CAR + cells at day 8. CAR T cells were cultured with GPC3 expressing HCC cells lines (Hep3B, HepG2, and Huh7) and GPC3 knockout (KO) cells at different effector to target ratios to assess specific lysis. (C) Study diagram if injection (i.p., 3 million), treatment of HCC tumors (i.v., 5 million), and blood collection in an NSG mouse model Intermediate and long hinges containing Fc are indicated by M or L after the construct name. (D) Bioluminescence imaging results quantifying tumor size. (E) Bioluminescence quantification of tumor size. Values represent mean±SEM. million *** p < 0.001; ns, not significant. Survival curve over full course of study (67 days). (F) PD1 indicating exhaustion shows that in all constructs at week 4. Standardized CAR T-cell counts and CAR-positive percentage in all groups at week 4. All listed subsets of T cells were also quantified at 4 and 10 weeks by flow cytometry. CAR, chimeric antigen receptor; PD1, programmed cell death protein 1.

Article Snippet: The cell surface binding the CAR on Jurkat cells to GPC3 protein tagged with hfc (ACRO Biosystems) was assessed using anti-human IgG-phycoerythrin (APC)-conjugated antibody (Jackson ImmunoResearch).

Techniques: In Vitro, In Vivo, Construct, Cell Counting, Transduction, Cell Culture, Expressing, Knock-Out, Lysis, Injection, Imaging, Flow Cytometry