Despite the success of checkpoint blockade immunotherapies, i.e., anti-PD1, only 20% of patients produce a durable response to these treatments. Thus, a need exists to develop additional therapeutic strategies to treat these patients, which includes evaluation of other tumor-infiltrating immune cells that could further augment the CD8+ and CD4+ T-cell response. Tumor-infiltrating B cells (TIL-B) represent a possible target for immunotherapy due to their predominance in the tumor microenvironment and crucial role in the immune response. In fact, current evidence suggests an antitumor role for TIL-Bs. Specifically, detection of TIL-Bs within tertiary lymphoid structures (TLS) correlates with both increased survival in patients with solid tumors and enhanced response to anti-PD1 immunotherapy. We hypothesize that TIL-Bs help generate potent, long-term immune responses against cancer by presenting tumor antigens to CD4 TILs within TLS. Utilizing single-cell RNAseq and high-level flow cytometry, we observed increased numbers of activated TIL-Bs in the primary tumors of head and neck squamous cell carcinoma (HNSCC) and non-small cell lung cancer (NSCLC) patients. We further assessed the location of TIL-Bs within the TME and determined that they predominantly sit adjacent to CD4+ tumor-infiltrating lymphocytes (TILs) within TLS. Thus, we generated an antigen presentation assay in vitro, and we observed increased CD4+ TIL responses when TIL-Bs presented autologous tumor antigens. Further, there was a differential impact on CD4+ TIL phenotype; specifically, if TIL-Bs were activated (CD27+), the CD4+ TILs were T helper (anti-tumor) CD4+ T cells and if the TIL-Bs were nonactivated (CD27-), the CD4+ TILs were T regulatory cells (protumor). These data suggest that TIL-Bs influence the phenotype and function of CD4+ TILs in patient tumors within TLS. Ultimately, results from this study will increase effective targeting of TIL-Bs within patient primary tumors.

Citation Format: Ayana T. Ruffin, Anthony R. Cillo, Sheryl Kunning, Robert L. Ferris, Dario A.A. Vignali, Tullia C. Bruno. Tumor-infiltrating B cells colocalize with CD4 T effector cells within tertiary lymphoid structures to present antigen and educate the antitumor immune response in human primary tumors [abstract]. In: Proceedings of the AACR-AHNS Head and Neck Cancer Conference: Optimizing Survival and Quality of Life through Basic, Clinical, and Translational Research; 2019 Apr 29-30; Austin, TX. Philadelphia (PA): AACR; Clin Cancer Res 2020;26(12_Suppl_2):Abstract nr PR06.