Objective: To investigate the potential diagnostic and prognostic values of antinuclear autoantibodies (ANAs) in diffuse large B-cell lymphoma (DLBCL). Materials and Methods: Eighty-two DLBCL patients and 120 healthy controls were selected from the Department of Hematology, Jincheng Dayi Hospital between 2005 and 2014. We examined the expression of ANA in the sera of the 82 DLBCL patients at different Ann-Arbor stages (15 at Stage I, 22 at Stage II, 27 at Stage III, and 18 at Stage IV). ANA detection was performed by immunofluorescence, and the results were confirmed by Western blotting analysis. Result: ANAs were more frequently detected in DLBCL patients than in controls (P < 0.001), with 25 (30.5%) DLBCL patients and 9 (7.5%) controls displaying elevated ANA levels. However, the majority of DLBCL patients in which ANA were detected did not develop autoimmune diseases, suggesting that ANA in DLBCL might not be correlated autoimmune diseases. Furthermore, no correlation was observed between the expression of ANA and the clinical stages of DLBCL. However, ANA-positive DLBCL patients had a better survival rate (P < 0.05). Conclusions: ANA in DLBCL may be a stage-independent prognostic factor rather than an indication for autoimmune diseases and may represent an effective immune response to the tumor.