PD-L1⁺ tumor-associated macrophages and PD-1⁺ tumor-infiltrating lymphocytes predict survival in primary testicular lymphoma

Primary testicular lymphoma is a rare and aggressive lymphoid malignancy, most often representing diffuse large B-cell lymphoma histologically. Tumor-associated macrophages and tumor-infiltrating lymphocytes have been associated with survival in diffuse large B-cell lymphoma, but their prognostic impact in primary testicular lymphoma is unknown. Here, we aimed to identify macrophages, their immunophenotypes and association with lymphocytes, and translate the findings into survival of patients with primary testicular lymphoma. We collected clinical data and tumor tissue from 74 primary testicular lymphoma patients, and used multiplex immunohistochemistry and digital image analysis to examine macrophage markers (CD68, CD163, and c-Maf), T-cell markers (CD3, CD4, and CD8), B-cell marker (CD20), and three checkpoint molecules (PD-L1, PD-L2, and PD-1). We demonstrate that a large proportion of macrophages (median 41%, range 0.08-99%) and lymphoma cells (median 34%, range 0.1-100%) express PD-L1. The quantity of PD-L1⁺ CD68⁺ macrophages correlates positively with the amount of PD-1⁺ lymphocytes, and a high proportion of either PD-L1⁺ CD68⁺ macrophages or PD-1⁺ CD4⁺ and PD-1⁺ CD8⁺ T cells translates into favorable survival. In contrast, the number of PD-L1⁺lymphoma cells or PD-L1^- macrophages do not associate with outcome. In multivariate analyses with IPI, PD-L1⁺ CD68⁺ macrophage and PD-1⁺ lymphocyte contents remain as independent prognostic factors for survival. In conclusion, high PD-L1⁺ CD68⁺ macrophage and PD-1⁺ lymphocyte contents predict favorable survival in patients with primary testicular lymphoma. The findings implicate that the tumor microenvironment and PD-1 - PD-L1 pathway have a significant role in regulating treatment outcome. They also bring new insights to the targeted thera py of primary testicular lymphoma.