|Budget Amount *help
¥3,300,000 (Direct Cost : ¥3,300,000)
Fiscal Year 2004 : ¥1,300,000 (Direct Cost : ¥1,300,000)
Fiscal Year 2003 : ¥2,000,000 (Direct Cost : ¥2,000,000)
Development of mice model of non-myeloablative stem cell transplantation for renal cell carcinoma using human cell line
A-498 cell line, which derived from human renal cell carcinoma, expresses the MUC-1 peptide on HLA-A2.
(1) Establishment of HLA-A2 restricted MUC-1 peptide specific cytotoxic T lymphocytes (CT L).
Adherent cells from human mononuclear cells obtained from normal volunteer with HIA-A2 were cultured with rhGM(1SF, rhIL-4, and TNF-a. These cells were stimulated by MUG-i peptide several times during the culture to get the dendric cells. In addition, mononuclear cells with HIA-A2 were added to the dendric cells with the several stimulations by MUC-1 peptide. After 7 days incubation, CTLs, which is specific cytotoxic against A-498 cells, were established.
(2) A498 bearing SCID mouse
A-498 cells were injected to SCID mice subcutaneously. The survival durations of A-498 bearing SCID mice were investigated. The survival durations in 5 mice with 9.0x10^6 A498 were 52,58,90,90 and 90days. Those in mice with 3.5x10^7 A498 were 30, 34, 70, 90 and 90days. Therefore, mice with 5.0x10^7 were used in the next experiments.
(3) in vivo anti-tumor effect of CTLs administrations on A-498 bearing SCID mice.
CTLs (1x10^6) were injected into A-498(5.0x10^7) bearing 5(111) mice through the tail vain once a week The survival durations were compared between A-498 bearing mice with the injections of (Ills and lymphocytes.
A-498 bearing mice with the injections of CTLs survived 42 and 90 days, while those with lymphocytes did 25 and 63 days
These results revealed that (Ills administration prolonged the survival duration of A-498 bearing SCUD mice. A-498 bearing SCID mice with the administrations of MUG-1 specific CTLs provide the mice model of non-myeloablative stem cell transplantation for renal cell carcinoma.