Diffuse Large B-cell Lymphomas Have Spatially-Defined Tumor-Immune Microenvironments Revealed by High-Parameter Imaging
Wright K*, Weirather JL*, Jiang S*, Kao KZ, Sigal Y, Giobbie-Hurder A, Shipp MA, Rodig SJ.
Diffuse large B-cell lymphoma, not-otherwise-specified, (DLBCL), is the most common aggressive non-Hodgkin lymphoma and a biologically heterogeneous disease. Despite the development of effective immunotherapies, the organization of the DLBCL tumor-immune microenvironment (TIME) remains poorly understood. We interrogated the intact TIME of 51 de novo DLBCLs with triplicate sampling to characterize 337,995 tumor and immune cells using a 27-plex antibody panel that captured cell lineage, architectural, and functional markers. We spatially assigned individual cells, identified local cell neighborhoods, and established their topographical organization in situ. We found that the organization of local tumor and immune cells can be modeled by six composite cell neighborhood types (CNTs). Differential CNT representation divided cases into three aggregate TIME categories: immune-deficient, dendritic-cell enriched (DC-enriched), and macrophage-enriched (Mac-enriched). Cases with immune-deficient TIMEs have tumor cell-rich CNTs wherein the few infiltrating immune cells are enriched near CD31-positive vessels in keeping with limited immune activity. Cases with DC-enriched TIMEs selectively include tumor cell-poor/immune cell-rich CNTs with high numbers of CD11c-positive dendritic cells and antigen-experienced T cells also enriched near CD31-positive vessels in keeping with increased immune activity. Cases with Mac-enriched TIMEs selectively include tumor cell-poor/immune cell-rich CNTs with high numbers of CD163-positive macrophages and CD8 T cells throughout the microenvironment, accompanied by increased IDO-1 and LAG-3 and decreased HLA-DR expression, and genetic signatures in keeping with immune evasion. Our findings reveal that the heterogenous cellular components of DLBCL are not randomly distributed but organized into CNTs that define aggregate TIMEs with distinct cellular, spatial, and functional features.