Fig. 1

Multiplex immunohistochemistry of cholangiocarcinomas. A Overlaid multiple images stained with immune cell markers with pseudocolors and selected immune cell subtypes defined by multiple myeloid cell marker were shown. B Percentages of immune cell subtypes among CD45⁺ cells. C–E Immune cell composition according to the average percentages of each type of immune cells among CD45⁺ cells in three different types of cholangiocarcinoma divided into the group of OS < 3 years and the group of OS > 3 years. The percentages of each cell type were compared between the groups of OS < 3 years and OS > 3 years by t test. *p < 0.05; **p < 0.01. F–K Kaplan–Meier curves of overall survival of patients whose tumors are grouped by higher density versus lower density of CD8⁺ T cells or CD8⁺granzyme B⁺ T cell subtypes as indicated. Log rank tests were done with p values indicated. DCC, distal cholangiocarcinoma; HC, hilar cholangiocarcinoma; ICC, intrahepatic cholangiocarcinoma