Immune checkpoint blockade (ICB) targeting PD-1/PD-L1 has been used for the treatment of head and neck squamous cell carcinoma (HNSCC). However, the overall response rate to ICB therapy for HNSCC remains less than 20%. It has recently been reported that the appearance of tertiary lymphoid structures (TLSs) in tumor tissue is correlated with better prognosis and response to ICB treatment. Here, we demonstrated an immune classification for the tumor microenvironment (TME) of HNSCC by analyzing The Cancer Genome Atlas (TCGA)–HNSCC data set and found that immunotype D with TLS enrichment had a better prognosis and response to ICB treatment. Furthermore, we observed that TLSs were present in a part of tumor samples of human papillomavirus (HPV) infection negative HNSCC (HPV⁻ HNSCC) and were associated with the densities of dendritic cell (DC)–LAMP⁺ DCs, CD4⁺ T cells, CD8⁺ T cells, and progenitor T cells in TME. We established an HPV⁻ HNSCC mouse model with TLS-enriched TME by overexpressing LIGHT in a mouse HNSCC cell line. We found that the induction of TLS formation enhanced the response to PD-1 blockade treatment in the HPV⁻ HNSCC mouse model, accompanied by increases in DCs and progenitor exhausted CD8⁺ T cells in the TME. Elimination of CD20⁺ B cells attenuated the therapeutic effect of PD-1 pathway blockade in TLS⁺ HPV⁻ HNSCC mouse models. These results indicate that TLSs contribute to the favorable prognosis and antitumor immunity of HPV⁻ HNSCC. Inducing TLS formation in HPV⁻ HNSCC tumors is a potential therapeutic method for improving the ICB response rate in patients with HPV⁻ HNSCC.

Cancer immunotherapy, which harnesses the patient's own immune system to target malignant cells, has shown remarkable promise in reducing tumor burden and extending survival. However, the complex tumor microenvironment (TME) limits therapeutic benefits to a subset of patients, making it challenging to develop accurate in vitro models for drug response prediction, drug discovery, and personalized medicine. Organoids, three-dimensional (3D) “mini-organs” derived from individual patients that faithfully recapitulate the structural, molecular, and gene expression profiles of primary tumors along with their complex TME in vitro, have emerged as powerful tools for patient-specific drug screening and therapeutic strategy development. Their versatility has led to widespread adoption across both clinical and basic cancer research. However, a key limitation of traditional organoid models is their lack of immune system components. Recent years have seen significant efforts to address this challenge through the integration of immune cells with organoids, aiming to create more physiologically relevant models. This review describes 3D culture methods for immunocompetent organoids, explores organoid–immune cell interactions, and discusses their applications in cancer immunotherapy and drug screening, along with recent advances in related clinical studies.