Immune Profiling and Quantitative Analysis Decipher the Clinical Role of Immune-Checkpoint Expression in the Tumor Immune Microenvironment of DLBCL.

Ziju Y Xu-Monette,Hua You,Govind Bhagat,Bing Xu,Lan V Pham,Nicholas Hoe,Eric D Hsi,Xiaohong Tan,Jane N Winter,Yong Li,Maurilio Ponzoni,Raghav Padmanabhan,Ganiraju C Manyam,Andrés J.M Ferreri,J Han Van Krieken,Min Xiao,Qingyan Au,Jing Wang,L Jeffrey Medeiros,Carlo Visco,George Z Rassidakis,Karen Dybkær,Jason R Westin,Ken H Young,Gordon J Freeman,Alexandar Tzankov,Wayne Tam,Hongwei Zhang,Yi Miao,Michael Möller ,Sandra Rodríguez‐Perales ,Miguel Á Piris ,Raúl Torres-Ruíz

doi:10.1158/2326-6066.cir-18-0439

Abstract

PD-1/L1 and CTLA-4 blockade immunotherapies have been approved for 13 types of cancers and are being studied in diffuse large B-cell lymphoma (DLBCL), the most common aggressive B-cell lymphoma. However, whether both PD-1 and CTLA-4 checkpoints are active and clinically significant in DLBCL is unknown. Whether PD-1 ligands expressed by tumor cells or by the microenvironment of DLBCL are critical for the PD-1 immune checkpoint is unclear. We performed immunophenotypic profiling for 405 patients with de novo DLBCL using a MultiOmyx immunofluorescence platform and simultaneously quantitated expression/coexpression of 13 immune markers to identify prognostic determinants. In both training and validation cohorts, results demonstrated a central role of the tumor immune microenvironment, and when its functionality was impaired by deficiency in tumor-infiltrating T cells and/or natural killer cells, high PD-1 expression (but not CTLA-4) on CD8+ T cells, or PD-L1 expression on T cells and macrophages, patients had significantly poorer survival after rituximab-CHOP (cyclophosphamide, doxorubicin, vincristine, and prednisone) immunochemotherapy. In contrast, tumor-cell PD-L2 expression was associated with superior survival, as well as PD-L1+CD20+ cells proximal (indicates interaction) to PD-1 + CD8+ T cells in patients with low PD-1 + percentage of CD8+ T cells. Gene-expression profiling results suggested the reversibility of T-cell exhaustion in PD-1+/PD-L1+ patients with unfavorable prognosis and implication of LILRA/B, IDO1, CHI3L1, and SOD2 upregulation in the microenvironment dysfunction with PD-L1 expression. This study comprehensively characterized the DLBCL immune landscape, deciphered the differential roles of various checkpoint components in rituximab-CHOP resistance in DLBCL patients, and suggests targets for PD-1/PD-L1 blockade and combination immunotherapies.

Highlights

Antitumor T-cell response is critical for the immune surveillance of cancer but is often damped by immune checkpoints
PD-L1 can interact with CD80 on activated T cells, and this interaction is required for T-cell anergy induction and maintenance [8]
Activated B-cell–like (ABC)-diffuse large B-cell lymphoma (DLBCL) compared with germinal center B-cell–like (GCB)-DLBCL had higher macrophage (P 1⁄4 0.0006) and CD8þ T-cell density (P 1⁄4 0.034; Supplementary Fig. S1A)

Summary

Introduction

Antitumor T-cell response is critical for the immune surveillance of cancer but is often damped by immune checkpoints. The checkpoint receptor CTLA-4 inhibits T-cell activation at the priming phase by competing with CD28 to bind CD80/ CD86 [1,2,3]. PD-1 suppresses the function of activated T cells through ligation of PD-1 ligands PD-L1 [4, 5] or PD-L2 [6], mainly at the effector phase [7]. The CTLA-4/PD-1 immune checkpoint can be reversed by CTLA-4/PD-1/PD-L1– blocking antibodies. The mechanisms of action are not completely understood [9, 10], and therapeutic effects of these blocking antibodies differ between CD4þ and CD8þ T cells [11, 12], between effector T and regulatory T cells Whether PD-L1 expressed by tumor cells or by host cells is essential for the immunotherapeutic efficacy of PD-1/L1 blockade remains controversial [9]

Methods

Results

Conclusion