There is an extensive search for natural products that can be introduced to broiler rations to improve performance, especially during the unfavorable breeding conditions. Under heat-stress conditions, the immune response seriously deteriorates, which consequently impairs broiler production performance. Thus, the present study aimed to investigate the potentials of Citrullus colocynthis seeds (CCs) supplementation to modulate the immune response of broilers subjected to chronic heat stress. A total of 300 Cobb-500 male broiler chickens aged 21 days were randomly divided into two equal groups and reared under either thermo-neutral condition (24 ± 1 °C) or subjected to cyclic heat stress (34 ± 1 °C for 8 h). Each group was further divided into two groups (5 replicate × 15 chicks) and was fed either the basal diet or the basal diet with 0.1% CCs supplementation. The results showed that heat stress impaired the production performance by lowering the final body weight and feed intake as well as impairing feed conversion. The levels of stress markers (i.e., malondialdehyde, tumor necrosis factor-α and corticosterone) increased (p < 0.05), whereas the activity of antioxidant enzymes decreased in broilers exposed to heat stress. Further, heat stress caused direct suppression of broiler humoral and cell-mediated immune responses. The stimulating index of T and B lymphocytes proliferation, as well as the antibody titer against sheep red blood cells, were significantly (p < 0.05) reduced by heat-stress exposure. However, CCs supplementation to broilers subjected to heat stress improved (p < 0.05) the final body weight, feed intake, and feed conversion ratio (FCR), compared to the non-supplemented stressed group. The cellular and cell-mediated immune response indicators significantly enhanced (p < 0.05) with CCs supplementation. Supplementation of CCs to broilers reared under similar environmental conditions elevated the total white blood cells (TWBCs) count and the broiler stimulating index of T and B lymphocytes. It can be concluded that CC seeds can be effectively used to stimulate the immune response and improve the production performance of broilers reared under heat-stress condition.