The results of our study show that: (a) differential intratumoral frequency of three T cell subsets, CD8+CD28+ T cells, CD8+CD28− T cells, and CD8+CD28−CD127−CD39+ Treg, distinguished between HNSCC patients who did or did not respond to treatment; (b) high PD-1 expression identified a CD8+CD28− T cell subpopulation phenotypically/functionally corresponding to CD8+CD28−CD127−CD39+ Treg; (c) CD8+CD28− T cell and CD8+CD28−CD127−CD39+ Treg subpopulations showed high expression of markers of exhaustion associated with regulatory function. Here, CD28 is linked to head and neck squamous cell carcinoma.