Beta-thalassemia is caused by a lack of or failure to synthesize beta globin chains in hemoglobin resulting in an excess of alpha chains. Cytokine-dependent hematopoietic cell linker (CLNK) is an adapter protein which is involved in the regulation of immunoreceptor signaling. It was found to be associated with a tyrosine-phosphorylated polypeptide (p92) in response to immunoreceptor stimulation. In thalassemia, oxidative stress causes tyrosine phosphorylation of the cytoplasmic domain of band 3. Therefore, we aimed to see how serum CLNK and interleukin-3 correlated with serum ferritin and annual transfusion index in children with beta-thalassemia major (b-TM). This case-control study included 100 non-splenectomized, transfusion-dependent b-TM pediatric patients receiving oral deferasirox and 100 healthy controls. The study was approved by the Institutional Review Board (IRB) of the Menoufia Faculty of Medicine, the approval number is 19/4/2021.PEDI. All procedures were carried out in accordance with relevant guidelines and regulations. In both groups, serum ferritin, interleukin-3, hemoglobin and CLNK levels were measured. They were found to be significantly higher in the b-TM patients than in the controls (p 0.001). There was a negative correlation between serum CLNK and hemoglobin (r = –0.483, p < 0.001), and a positive correlation between serum CLNK and ferritin levels (r = 0.855, p < 0.001). There was a positive correlation between serum CLNK, ferritin, and annual transfusion index. Increased serum CLNK in transfusion-dependent b-TM patients was associated with elevated serum ferritin concentrations and high annual transfusion index. This could be explained by reciprocal effects between immune signaling system and immature erythrocytes which release signaling molecules, such as CLNK, in the blood.