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bcr-ablペプチドをパルスしたdendritic cell を用いたbcr-ablペプチド特異的自己細胞障害性T細胞の誘導： 高橋益広¹、オスマン・ヤーセル²、鄭　智茵²、劉　愛春²、古川達雄²、鳥羽　健²、青木定夫²、小池　正²、相沢義房²（新潟大・¹医療短大、²第一内科）

Generation of bcr-abl peptide specific autologous　 cytotoxic T cells by using dendritic cells pulsed with bcr-abl peptide: Masuhiro TAKAHASHI¹, Yasser OSMAN², Zhiyin ZHENG², Aichun LIU², Tatsuo FURUKAWA², Ken TOBA², Sadao AOKI², Tadashi KOIKE², Yoshifusa AIZAWA² (Clg. of Biomed. Tech., Niigata Univ., 1st Dept. of Int. Med., Niigata Univ.)

In order to establish an efficient anti-tumor immunotherapy using dendritic cells (DC) presenting tumor specific antigen, we studied the effect of DC pulsed with bcr-abl peptide on generation of bcr-abl peptide specific autologous cytotoxic T lymphocytes (CTL). DC were generated by culturing normal blood adherent cells or purified blood DC precursors (CD3^-, CD11b^-, CD16^-, CD4⁺) with GM-CSF (100 ng/ml) and IL-4 (10 ng/ml). On day 3 and 5 of culture, DC were pulsed with bcr-abl (b3-a2) fusion peptide (GFKQSSKALQRP) (25 ng/ml) and on day 6 TNF-α (10 ng/ml) was added to the culture.
On day 12, 30 Gy irradiated DC (2x10⁵) were cocultured with autologous blood non-adherent cells (2x10⁶) in 2 ml medium for 6 days. Blood adherent cells, which were pulsed with b3-a2 peptide on day 3, were cultured with M-CSF (50 ng/ml) for 6 days and used as target cells for cytotoxicity test.
Autologous CTL induced by pulsed DC were cocultured with ⁵¹Cr labeled target macrophages at various effector:target (E:T) ratio and ⁵¹Cr-release was evaluated. Although more than 50% of pulsed target cells were killed by CTL induced by pulsed DC at the E:T ratio of 30:1, non-pulsed target cells were not killed in the system using autologous plasma. These results demonstrated that DC pulsed with antigenic peptide can generate antigen specific CTL and suggested the applicability of tumor specific antigen pulsed DC for tumor immunotherapy.