Cyclophosphamide (CTX) is a chemotherapeutic drug used to treat mammary gland tumors. Despite its cytotoxic effect on cancer cells, it causes harm to hepatic and renal tissues, ranging from mild infiltration to necrosis and finally cytolysis, depending on the dosage and duration of treatment. It also suppresses bone marrow, resulting in leukopenia, anemia, and thrombocytopenia. This study aimed to evaluate the protective effects of concanavalin A (con-A) and IL-27 conditioned immune therapeutics on Ehrlich ascites carcinoma (EAC) tumor-bearing mice treated with CTX. To achieve this, naive splenocyte cells were activated in vitro for 3 days with the T cell mitogen concanavalin A (Con-A; 5 µg/mL) and/or IL-27 (10; ng/mL). After a single i/p injection of CTX (4 mg/mouse), they were adoptively transferred into (EAC) tumor-bearing mice. After tumor implantation, hematological analysis, liver function, and kidney function were evaluated. According to the findings, the CTX-treated group had significantly lower total leucocyte count, lymphocytes, neutrophils, hematocrit, and RBCs than the control group. Interestingly, Con-A + IL-27 normalized the total leucocytes' count, especially lymphocytes and neutrophils. When Con-A + IL-27 was compared to CTX, there was a significant increase in hematocrit and platelet count. AST, ALT, bilirubin, creatinine, and urea levels were not significantly different between normal and Con-A + IL-27, but CTX significantly increased AST compared to normal. According to the findings, adoptive transfer of Con-A + IL-27 conditioned splenocyte cells into lymphopenic hosts restored hematological and biochemical parameters and recovery from chemotherapy-induced lymphopenia.