Human papillomavirus-associated immune profile of the tumor microenvironment in laryngeal squamous cell carcinoma: a Tunisian cohort study

Abstract Background Human papillomavirus (HPV) infection plays a key role in the pathogenesis of several squamous cell carcinomas and may influence tumor immune responses. Although HPV-associated oropharyngeal squamous cell carcinoma has been extensively studied, the relationship between high-risk HPV (HR-HPV) status and the tumor immune microenvironment (TME) in laryngeal squamous cell carcinoma (LSCC) remains poorly understood. This study aimed to investigate the association between HR-HPV status and immune cell infiltration in LSCC. Methods A retrospective cohort of 70 patients with histologically confirmed LSCC was analyzed. HR-HPV DNA detection was performed on tumor specimens. Immunohistochemical analysis was used to evaluate CD8⁺ cytotoxic T cells, FOXP3⁺ regulatory T cells, and CD68⁺/CD163⁺ tumor-associated macrophages. Immune infiltration was assessed using a semi-quantitative immunoreactivity score. Associations between immune markers, HR-HPV status, clinicopathological parameters, and survival outcomes were evaluated. Results HR-HPV DNA was detected in 39/70 (55.7%) LSCC cases. HR-HPV-positive tumors showed significantly increased infiltration of CD8⁺ T cells, FOXP3⁺ Tregs, and CD68⁺/CD163⁺ macrophages compared with HR-HPV-negative tumors (p < 0.0001). High CD8⁺ T-cell infiltration was associated with improved overall and disease-free survival, although without statistical significance. FOXP3⁺ Treg infiltration was significantly associated with tumor grade. Conclusions HR-HPV-positive LSCC exhibits a distinct immune landscape characterized by increased infiltration of T cells and tumor-associated macrophages, suggesting HPV-driven immune modulation within the TME. These findings highlight the importance of immune profiling in HPV-related LSCC and support further studies exploring its prognostic and therapeutic implications.