Tumor-infiltrating B cells produce tumor-specific antibodies and may contribute to suppressing tumor in head and neck squamous cell carcinoma.

The involvement of T cells, particularly cytotoxic T cells, in tumor immunity has been reported, but the roles of B cells and antibodies in tumor immunity remain unclear. Previous studies have primarily focused on circulating B cells in the blood, with limited investigation of tumor-infiltrating B cells (TIL-Bs) and their secreted antibodies in head and neck squamous cell carcinoma (HNSCC) tissue. This study aimed to clarify TIL-Bs' role in HNSCC through comprehensive analyses including single-cell RNA sequencing of 68 cases, B-cell receptor repertoire analysis, multiplex immunofluorescence staining, and spatial transcriptomics analysis of resected HNSCC specimens. We identified CXCL13⁺CD4⁺ T cells that attract B cells, along with TIL-Bs at various differentiation stages, showing clonal expansion of antibody-secreting cells (ASCs). These ASCs exhibited a low somatic hypermutation frequency, indicating that they are likely activated through extrafollicular response rather than germinal center response. Protein microarrays with recombinant antibodies mimicking TIL-Bs secretory antibodies and serum antibodies revealed several antigens highly expressed in tumor cells. These antigens demonstrated higher expression in tumor cells than normal epithelial cells in the resected HNSCC specimens. Tissue analysis indicated that antibodies binding to these antigens are recognized by macrophage and NK cells with Fc receptors, potentially enhancing tumor immunity. The existence of tumor-infiltrating ASCs showed correlation with favorable prognosis. This study demonstrated for the first time that expanded TIL-Bs produce antibodies and likely contribute to tumor elimination by activating tumor immunity in HNSCC tissues. This novel insight could lead to radically new therapies targeting B cells.