Fig. 4: PDL1 expression of macrophages is associated with shorter progression-free survival and is mutually exclusive with Treg infiltration.

A–F The “amplicon-sequenced DLBCL” TMA was stained with antibodies against CD8, PD1, CD68, PDL1, TCF1 and CD79a and DAPI (D). Normal lymph node samples were stained with the same panel for comparison. Representative images of a normal lymph node, and of a representative DLBCL case are shown at low and high (inset) magnification in (A, B) (note that the tumor/B-cell marker CD79a is not shown for the sake of clarity). A gallery of single and multi-channel images of four macrophage populations is shown in (C). Asterisks in A–C indicate cells of interest. The relative abundance (in % of all cells with a DAPI⁺ nucleus) of the eight numerically dominant macrophage and CD8⁺ T-cell populations, plus marker-negative populations (“other”, also including PDL1⁺ CD68^- cells), is shown in D in the form of stacked bar plots for all cases included on the TMA. Patients are arrayed on the x-axis and sorted by COO classification into ABC- and GCB-DLBCL. The relative abundance (in % of all cells with a DAPI⁺ nucleus) of the indicated populations as detected in the two COO subgroups is shown in (E) (macrophage populations on the left; CD8⁺ T-cell populations on the right). The relative abundance of the indicated macrophage populations, as detected in the five genetic subtypes of DLBCL (determined using the 2-step classifier; NA not annotated), is shown in (F); each symbol represents one case in (E, F). Statistical comparisons in E and F were performed by one-way ANOVA with Dunnett’s T3 correction; *p < 0.05; **p < 0.01. G Average number of cells, of the indicated populations, that reside within a 10-pixel radius of a PDL1⁺ CD68⁺ macrophage. Each symbol represents one of the amplicon-sequenced DLBCL patients; there are 10 dots per patient, one for each of the indicated populations. Other PDL1⁺ CD68⁺ macrophages, and tumor cells are the most common neighbors of PDL1⁺ CD68⁺ macrophages in DLBCL. H Kaplan-Meier curve showing progression-free survival of patients with large (>median) relative to small (<median) infiltrates of CD68⁺ PDL1⁺ macrophages; the cohort plotted here comprises the 77 DLBCL patients included in the NHL TMA and was stained with (D). I Kaplan-Meier curve showing progression-free survival of the same 77 DLBCL patients with > or <median infiltrates of CD68⁺ PDL1⁺ macrophages and CD8⁺ T-cells. The p-values in H and I were calculated by Kaplan-Meier log-rank test. J, K Relative abundance of Tregs, vs. relative abundance of PDL1⁺ macrophages (each in % of cells with a DAPI⁺ nucleus), of the amplicon-sequenced DLBCL cohort (in J) and the NHL cohort (DLBCL patients only; K). The color code indicates COO. L Representative micrographs of sections stained for the indicated markers, of cases of which one is highly infiltrated by PDL1⁺ macrophages but not Tregs (left panels, UPN016) and one is highly infiltrated by Tregs but not PDL1⁺ macrophages (right panels, UPN216).