BackgroundThe presence and activity of tumor-infiltrating lymphocytes (TILs) is a key parameter related to the antitumor immune response. A large number of studies reveal TIL density as a prognostic marker and predictor of response to radiotherapy, chemotherapy, and immunotherapy.MethodsWe examined the TIL and tertiary lymphoid structure TLS density in the invading front and inner tumor stroma, in a 33 squamous cell laryngeal carcinomas (LSCC) treated with laryngectomy. TIL and TLS densities were in parallel examined with markers of anaerobic metabolism, vascular density (VD), vascular survival ability (VSA), and histopathological parameters.ResultsTIL and TLS densities significantly decreased in inner tumor areas (p< 0.0001). TIL density in the invading tumor front was inversely related with lymph node involvement (p= 0.03), HIF1α expression (p= 0.008), vessel density (p= 0.02), and MIB1 (p= 0.006). TIL density in inner stroma was inversely linked to local invasion (marginalp= 0.05), tumor budding (TB) (p= 0.005), MIB1 (p= 0.02), and HIF1α expression (p= 0.02). Low-TLS density in the invading front and in inner tumor areas was related to high TB (p= 0.02 and 0.002, respectively), HIF1α (p= 0.003 and 0.01, respectively), and LDH5 expression (p= 0.003 and 0.007, respectively). CD4+, FOXP3+TIL density, and FOXP3+/CD8+ratio were directly associated with VSA (p= 0.008, 0.02, and 0.05, respectively).ConclusionPoor immune response is related to hypoxic background and anaerobic metabolism, as well as increased invasive and metastatic ability. Regulatory TIL markers are linked with increased angiogenic potential. The prognostic, predictive, and therapy-guiding value of TILs in clinical practice demands thorough investigation.Supplementary InformationThe online version contains supplementary material available at 10.1007/s12105-022-01517-7.