Tumor-Infiltrating Plasma Cells Are Associated with Tertiary Lymphoid Structures, Cytolytic T-Cell Responses, and Superior Prognosis in Ovarian Cancer

Abstract

Purpose: CD8+ tumor-infiltrating lymphocytes (TIL) are key mediators of antitumor immunity and are strongly associated with survival in virtually all solid tumors. However, the prognostic effect of CD8+ TIL is markedly higher in the presence of CD20+ B cells, suggesting that cooperative interactions between these lymphocyte subsets lead to more potent antitumor immunity. Experimental Design: We assessed the colocalization patterns, phenotypes, and gene expression profiles of tumor-associated T- and B-lineage cells in high-grade serous ovarian cancer (HGSC) by multicolor IHC, flow cytometry, and bioinformatic analysis of gene expression data from The Cancer Genome Atlas. Results: T cells and B cells colocalized in four types of lymphoid aggregate, ranging from small, diffuse clusters to large, well-organized tertiary lymphoid structures (TLS) resembling activated lymph nodes. TLS were frequently surrounded by dense infiltrates of plasma cells (PC), which comprised up to 90% of tumor stroma. PCs expressed mature, oligoclonal IgG transcripts, indicative of antigen-specific responses. PCs were associated with the highest levels of CD8+, CD4+, and CD20+ TIL, as well as numerous cytotoxicity-related gene products. CD8+ TIL carried prognostic benefit only in the presence of PCs and these other TIL subsets. PCs were independent of mutation load, BRCA1/2 status, and differentiation antigens but positively associated with cancer–testis antigens. Conclusions: PCs are associated with the most robust, prognostically favorable CD8+ TIL responses in HGSC. We propose that TLS facilitate coordinated antitumor responses involving the combined actions of cytolytic T cells and antibody-producing PCs. Clin Cancer Res; 22(12); 3005–15. ©2016 AACR.

High Density of Tumor-infiltrating B-Lymphocytes and Plasma Cells Signifies Prolonged Overall Survival in Adenocarcinoma of the Esophagogastric Junction

“…Increased plasma cell density has been reported to be related to improved outcome in non-small cell lung cancer (NSCLC) and high-grade ovarian carcinoma (7,21), while other studies showed an association with higher tumor grade and reduced OS in ovarian cancer (22).…”

Tertiary Lymphoid Structures Associate with Tumour Stage in Urothelial Bladder Cancer

“…Type 4 aggregates contain very few T-cells, but have well-formed GCs containing CD20 + B cells and CD21 + fDCs. In our study, we defined TLS as lymphoid aggregates with well-formed GCs, which included Type 3 and Type 4 lymphoid aggregates similar to those described in ovarian cancer by Kroeger and colleagues [21].…”

“…1). We followed the recently reported classification criteria for lymphoid aggregates based on size, cellular composition and degree of organization to define the types of TLS in the cohort under study [21]. Type 1 aggregates are small, compact, T-cell predominant lymphoid aggregates without apparent follicular structures.…”

“…To further characterize the lymphoid structures observed in H&E stained sections, we used a previously described [21] 6-colour IHC panel that enabled simultaneous detection of CD20 + B cells, CD3 + and CD8 + T cells, CD21 + fDC, CD208 + activated conventional DC, and PNAd + high endothelial venule (HEV)-like vessels. All TLS identifiable as lymphoid aggregates by H&E were confirmed by IHC to contain well-defined follicular structures with GC highlighted by CD20 + B cells and CD21 + fDC networks surrounded by a varying number of CD3 + and CD8 + T-cells.…”

“…Multicolour IHC was performed as previously described by Kroeger and colleagues [20,21] at the Deeley Research Center (BC cancer agency, Canada). IHC was performed on 5m thick FFPE sections as previously described [20,21] on the Intellipath FLX Automated Staining system, using mouse antihuman CD20 (1 : 500 dilution, clone L26, Biocare Medical), rabbit anti-human CD3 (1 : 250 dilution, clone SP7, Spring Biosciences), mouse anti-human CD8 (1 : 300 dilution, C8/144B, Cell Marque), rat anti-human CD208 (1 : 100, clone 1010E1.01, Novus Biologicals), rat anti-human PNAd (1 : 100 dilution, MECA-79, Biolegend) and mouse anti-human CD21 (1 : 100 dilution, clone 2G9, Biocare Medical) antibodies. Multispectral images of stained sections were captured using the PerkinElmer Vectra system.…”