Effective immunotherapy options for patients with non–small cell lung cancer (NSCLC) are becoming increasingly available. The immunotherapy focus has been on tumor-infiltrating T cells (TILs); however, tumor-infiltrating B cells (TIL-Bs) have also been reported to correlate with NSCLC patient survival. The function of TIL-Bs in human cancer has been understudied, with little focus on their role as antigen-presenting cells and their influence on CD4⁺ TILs. Compared with other immune subsets detected in freshly isolated primary tumors from NSCLC patients, we observed increased numbers of intratumoral B cells relative to B cells from tumor-adjacent tissues. Furthermore, we demonstrated that TIL-Bs can efficiently present antigen to CD4⁺ TILs and alter the CD4⁺ TIL phenotype using an in vitro antigen-presentation assay. Specifically, we identified three CD4⁺ TIL responses to TIL-Bs, which we categorized as activated, antigen-associated, and nonresponsive. Within the activated and antigen-associated CD4⁺ TIL population, activated TIL-Bs (CD19⁺CD20⁺CD69⁺CD27⁺CD21⁺) were associated with an effector T-cell response (IFNγ⁺ CD4⁺ TILs). Alternatively, exhausted TIL-Bs (CD19⁺CD20⁺CD69⁺CD27⁻CD21⁻) were associated with a regulatory T-cell phenotype (FoxP3⁺ CD4⁺ TILs). Our results demonstrate a new role for TIL-Bs in NSCLC tumors in their interplay with CD4⁺ TILs in the tumor microenvironment, establishing them as a potential therapeutic target in NSCLC immunotherapy. Cancer Immunol Res; 5(10); 898–907. ©2017 AACR.