A multiparametric analysis of T cell infiltration in tumor biopsies using IHC, MAGE-A4 SPEAR RNAscope (Advanced Cell Diagnostics) and multiplex immunofluorescence. Images of post-infusion biopsy (liver) from Patient 4: A, spatial plot generated using spatial analysis module in HALO (Indica Labs) showing malignant cells (PanCK + (red), regulatory (CD3 + CD4 + FoxP3 + ) T cells (yellow) and cytotoxic (CD3 + CD8 + ) T cells (blue/cyan)). Scale bar is not applicable as this is not a raw image; B, CD3 IHC/SPEAR + T cell RNAscope duplex (CD3 IHC staining (blue); MAGE-A4 SPEAR T cell staining (purple)); C, Ultivue 8-plex multiplex dataset showing CD4 (orange), PD-L1 (red), CD8 (green), Ki67 (purple), FoxP3 (blue), GrazB (white), CD3 (yellow) and PanCK (teal); D, PanCK immunofluorescence staining from Ultivue 8-plex panel (displayed in absorption mode in HALO for clarity); E, MAGE-A4 IHC stain (DAB (brown)); and F, hematoxylin (purple) and eosin (pink) stain.
