Abstract Understanding the impact of tertiary lymphoid structures (TLSs) on acral melanoma (AM) and the tumor microenvironment (TME) is critical. We analyzed TLS features in primary AM lesions from 46 patients and identified intratumoral TLSs (intra-TLSs) in 25 patients. Intra-TLS presence was significantly associated with improved overall survival. Hematoxylin and eosin staining and multiplex immunofluorescence revealed increased T-cell and CD8+ T-cell infiltration and fewer tumor-associated macrophages in the TME of intra-TLS patients. Transcriptomic analysis identified a TLS-associated Th1/B-cell gene set as a predictor of survival and immunotherapy response. These findings highlight the prognostic value of intra-TLSs in AMs and suggest that targeting TLS formation could enhance immunotherapy efficacy.