Objective <p>To investigate the clinical characteristics and survival of p53 and Programmed death-ligand 1 (PD-L1) co-expression in diffuse large B-cell lymphoma (DLBCL) patients.</p> Methods <p>Immunohistochemistry (IHC) was used to detect the expression of p53 and PD-L1 in tumor cells of 176 patients with DLBCL. Clinical data were retrospectively examined along with long-term follow-up. Correlation between the expression of p53 and PD-L1 and the clinicopathological characteristics of the patients was assessed. Impact of Rituximab (R) on prognosis among DLBCL patients with positive expression of p53 and PD-L1 was evaluated.</p> Results <p>p53 expression, PD-L1 expression, and p53/PD-L1 co-expression were present in 44.9%, 42.0%, and 25.6% of patients with DLBCL, respectively. No significant differences existed in the clinicopathological characteristics between patients with positive and negative p53 expression and between patients with positive and negative PD-L1 expression. However, more co-expression of p53/PD-L1 was observed in patients with non–germinal center B-cell-like (non-GCB) subtypes (<i>p</i> = 0.005). There was a significant positive correlation between p53 and PD-L1 expression (<i>r</i> = 0.273, <i>p</i> &lt; 0.001). Survival analysis indicated that patients exhibiting p53/PD-L1 co-expression had decreased progression-free survival (PFS) and overall survival (OS ) compared to those demonstrating positive expression of either p53 or PD-L1, and in addition, both negative p53/PD-L1 expression. Rituximab failed to substantially modify the prognosis of patients. p53/PD-L1 co-expression is an independent indicator of adverse prognosis.</p> Conclusions <p>Co-expression of p53/PD-L1 in DLBCL patients implies an unfavorable prognostic group, which doesn’t derive benefit from Rituximab.</p>