Thermal response and hyperthermic radiosensitization of scid mouse bone marrow CFU-C

Abstract

Purpose : Scid mice are severely immunodeficient as a result of a defective recombinase system. Mice with the scid mutation have been shown to have an increased sensitivity to ionizing radiation, presumably as a result of an inability to repair DNA damage. Little is known of the impact of this mutation on the thermal response and on hyperthermic radiosensitization. This investigation established the thermal response (42–44°C), patterns of thermotolerance development, and the impact of hyperthermia (60 min at 40°C or 42°C) on the radiation response of bone marrow colony forming unit—culture cells (CFU-C) in scid mice. Methods and Materials : Anesthetized scid mice (pentobarbital, 90 mg/kg) were killed by cervical dislocation and the nucleated marrow obtained from both tibia and femora by passing 2 ml of cold McCoy's 5A medium supplemented with 15% fetal bovine serum through each bone. Single cell suspensions of nucleated marrow were heated in 12 × 75 mm sterile tissue culture tubes at a concentration of approximately 5 × 10 6 cells/ml. Radiation, when used, was delivered immediately prior to hyperthermia by a 137Cs irradiator (dose rate of 1.20 Gy/min). Colony forming unit—culture were cultured in semisolid agar in the presence of colony stimulating factor (conditioned medium from L929 cells) for 7 days. Results : The slope of the radiation dose—response curve for CFU-C in scid mice was biphasic, the Dos (± SE) were 0.29 ± 0.03 Gy and 1.09 ± 0.20 Gy, respectively. The Dos of the radiation dose—response curve for wild type marrow from CB-17 and Balb/c mice were 1.28 ± 0.05 Gy and 1.47 ± 0.15 Gy, respectively. The Dos of the hyperthermia dose—response curves for scid mice were 75 ± 5, 10 ± 1.4, and 4 ± 0.2 min, respectively, for temperatures of 42°, 43°, and 44°C. Thermotolerance development at 37°C increased to a maximum at approximately 240 min after acute hyperthermia (15 min at 44°C) and thereafter, decreased to control levels within 15 h. Thermotolerance did not develop in scid CFU-C during chronic hyperthermia at temperatures < 42.5°C. Hyperthermia (60 min at 40° or 42°C) immediately after ionizing radiation did not significantly alter the terminal slope of the radiation dose—response curve of scid CFU-C (Do = 1.28 ± 0.08 Gy). By contrast, hyperthermia following radiation of wild type CFU-C resulted in a decrease in the Do from 1.47 ± 0.05 Gy (Balb/c, rad only) to 1.31 ± 0.08 or 1.06 ± 0.18 Gy for 60 min at 40° or 42°C, respectively. Conclusion : These results show that the thermal response and the pattern of thermotolerance development of scid CFU-C were similar to that of wild type Balb/c CFU-C, but that hyperthermia given immediately after ionizing radiation did not alter the radiation response of scid CFU-C. The scid mutation does not increase hyperthermic sensitivity or change the pattern of thermotolerance development of scid mouse CFU-C, implying that the scid mutation is not involved with thermal response, but does render the already radiation-sensitive scid cells incapable of thermal radiosensitization.