Fig. 3

Tumor and blood immunologic analyses. a Pre-treatment tumor biopsy and post-treatment tumor and gastrointestinal (GI) biopsies from patient 8 showing expression of macrophages in different stages of differentiation (shown in red, purple and orange due to expression of different combinations of markers), and natural killer (NK) cells (green) in column A, cytotoxic T lymphocytes (CTL; blue/purple), helper T cells (Thelper; green/orange) and immature thymocytes (white) in column B, plasma cells (green) in column C, and B cells (red/orange) in column D. Higher macrophage, NK cell and CTL expression was seen in tumor samples after treatment. Scattered plasma cells were observed with no appreciable change after treatment. No significant B cell population was observed, except in one field of view of the GI biopsy (shown above). b Post-treatment tumor biopsy from a lesion demonstrating response from patient 3 showing expression of macrophages (red, blue, purple and orange) in Panel A, natural killer (NK) cells (green) in Panel B, and cytotoxic T lymphocytes (CTL, blue/purple) in Panel C. c Pre-treatment tumor biopsy and post-treatment tumor and gastrointestinal (GI) biopsies from patient 8 after nine doses of avelumab showing increased expression of HLA I (red/pink/purple due to overlap with pan-leukocyte marker in blue, or orange due to overlap with pan-cytokeratin marker in green; column A), low and heterogeneous expression of HLA II (green) surrounded by macrophages (red/blue/purple) (column B) in post-treatment samples. Regulatory T cells (Tregs) were not present in significant numbers in pre- and post-treatment samples (column C). d Absolute lymphocyte count (ALC) and frequency of immune cell subsets prior to therapy (baseline) that are differentially expressed between clinical responders (patients 1, 3, 6, and 8) and non-responders (patients 2, 4, 5, 7). Patient 2 (a clinical non-responder who developed autoimmune adverse event like the responders) is noted with an open square. cDC, conventional dendritic cells