Fig. 5: IMC-KRASG12D mediated T cell activation and redirected killing of cancer cells expressing KRASG12D, but not KRASWT.
From: Therapeutic high affinity T cell receptor targeting a KRASG12D cancer neoantigen
a Immature dendritic cells differentiated from monocytes isolated from healthy donors were transfected with mRNA encoding KRASWT or KRASG12D or b pulsed with indicated exogenous peptide, were treated with IMC-KRASG12D and co-cultured with autologous T cells for 24 h. T cell activation was measured by IFNγ ELISPOT assay. Three biological replicates from one representative experiment from two independent experiments are shown. c Immunoblot and densitometry analysis of isogenic PSN-1 cell lysates. Blot is representative of two independent experiments. Uncropped blots can be viewed in Supplementary Fig. S10. d IFNγ ELISPOT output of cell lines modified to express HLA-A*11 and KRASG12D (clones 1 & 2), HLA-A*11 and KRASWT (clone 3) or only KRASG12D (clone 4) treated with IMC-KRASG12D and co-cultured with PBMC for 24 h. Targets alone, or no IMC-KRASG12D negative controls (Cont.) were performed. Mean data of three biological replicates ±SEM from one representative experiment from three independent experiments is shown. e IFNγ ELISPOT output of an extended panel of HLA-A*11-KRASG12D + and HLA-A*11-KRASG12D- cancer cell lines and healthy cells, treated with IMC-KRASG12D and co-cultured with PBMC for 24 h. HLA-A*11 + /KRASG12D- cell lines were pulsed with 10 µM KRASG12D (‘+G12D’) peptide as a positive control. Mean data of three biological replicates ±SEM from one representative experiment from two independent experiments is shown f. Redirected T cell killing assay of HLA-A*11 + /KRASG12D CL40 cancer or HLA-A*11 + /KRASWT normal human colonic epithelial cells expressing nuclear-restricted mKATE2, treated with IMC-KRASG12D and co-cultured with PBMC for 72 h. Cells were pulsed with 10 µM KRASG12D (‘+G12D’) peptide as a positive control. Targets alone, PBMC alone or no IMC-KRASG12D negative controls (Cont.) were performed. Mean data of three biological replicates ±SD from one representative experiment from two independent experiments is shown. Source data are provided as a Source Data file.
