Induction of antitumor immunity to T-9 rat glioma by intracellular hyperthermia using functional magnetic particles was investigated. Magnetite cationic liposomes (MCLs), which have a positive surface charge, were used as heating mediators for intracellular hyperthermia. Solid T-9 glioma tissues were formed subcutaneously on both femurs of female F344 rats, and MCLs were injected via a needle only into the left solid tumors (treatment side). The rats were then divided into two groups, which received no irradiation, or irradiation for 30 min given three times at 24-h intervals with an alternating magnetic field (118 kHz, 384 Oe). On the treatment side, the tumor tissue disappeared completely in many rats exposed to the magnetic field. The tumor tissue on the opposite side also disappeared completely, even though MCLs were not injected into the right solid tumors. To examine whether a long-lasting and tumor-specific immunity could be generated, the rats that had been cured by the hyperthermia treatment were rechallenged with T-9 cells 3 months later. After a period of transient growth, all tumors disappeared. Furthermore, immunocytochemical assay revealed that the immune response induced by the hyperthermia treatment was mediated by both CD8 Various methods have been employed, such as whole body hyperthermia, 2) radiofrequency hyperthermia, 3) and inductive hyperthermia using microwave antenna 4) or implantable needles. 5) However, it is always difficult to achieve uniform heating of the tumor region to the desired temperature without damaging normal tissue. Therefore, some researchers have proposed intracellular hyperthermia and developed submicron magnetic particles for this purpose.6-8) These magnetic particles are easily incorporated into cells and generate heat under an alternating magnetic field through hysteresis loss. 9) We have also developed 'magnetite cationic liposomes' (MCLs) for intracellular hyperthermia.10, 11) MCLs were developed to improve adsorption and accumulation into the tumor cells and show ten-fold higher affinity for tumor cells than neutrally charged magnetoliposomes, 10) owing to electrostatic interaction with the negatively charged cell membrane. 12,13) The hyperthermic effect of the MCLs was examined in vivo.14) MCLs were injected into solid tumors formed subcutaneously in F344 rats and the rats were irradiated three times for 30 min with an alternating magnetic field. Histological observations were carried out just after the irradiation and showed that some tumor cells survived, especially in the peripheral area. However, complete tumor regression was observed one month after the irradiation. We were interested in the possibility that antitumor immunity had been induced by the hyperthermic treatment using MCLs.In the present paper, it is demonstrated that our hyperthermia system can induce an antitumor immune response and the acquired immunity is long-lasting.
MATERIALS AND METHODS
MaterialsDilauroylphosphatidylcholine and dioleoylphosphatidylethanolamine were purchased from Sigma Ch...