Background: Unintentional irradiation of the spleen may reduce absolute lymphocyte count (ALC), which can affect tumor immunity. Therefore, in the present study, we evaluated spleen dose-volume parameters associated with ALC and neutrophil/lymphocyte ratio (NLR) in patients with esophageal cancer.Methods: This retrospective study evaluated patients who were diagnosed with stage I-IV esophageal cancer and who received radiotherapy at Nihon University Itabashi Hospital between January 2015 and March 2020. Spleen V5, V10, V20, and V30 and mean spleen dose, which are dose-volume histogram parameters correlated with ALC and NLR, were analyzed.Results: In total, 89 esophageal cancer patients with a median age of 72 years (range, 51-92 years) were included in this research. Results showed that spleen V5, V10, and V20 and mean splenic dose were significantly correlated with a low ALC. Meanwhile, a significant association was observed between spleen V5 and V10 and a high NLR (P<0.05). In the linear regression analysis, spleen V5 and V10 were remarkably associated with a low ALC (P=0.006 and 0.008). Further, a correlation was noted between spleen V5 and a high NLR (P=0.019). Spleen V5 and V10 were remarkably associated with greater than grade 3 lymphopenia (P=0.024 and P=0.031).Conclusions: A correlation was observed between the irradiated volume of the spleen and ALC and NLR in patients with esophageal cancer. Furthermore, the spleen should be regarded as a high-risk organ, and the use of techniques in reducing spleen V5, V10 for the preservation of tumor immunity may be beneficial.