| Objectives: To provide technic methods and experimental evidences for clinical application of tumor biological therapy associated with the sensitive dendritic cells (DCs).Methods: The DCs obtained from long bone marrow of female mice were amplified with granulocyte macrophage colony stimulating factor(GM-SCF)and interleukin-4(IL-4) in vitro. Animal models were established by being inoculated subcutaneously (s.c) with melanoma cells. Bearing tumor mice were treated with tumor lysate-pulsed DCs for immunoprophylaxis and immunotherapy. The protection and therapy effects of sensitive DCs to the bearing tumor mice were observed, and levels of the mice serum TNF-α,IL-6 were detected with ELISA technique.Results:1. The bearing tumor rate of mice inoculated with 5×105 B16 cells was 100%, and the rate of mice inoculated with 5×104 B16 cells was 60%. Average surviving time of the latter was significantly longer than that of the former (P<0.01). 2. The average surviving time and tumor volum were not significantly different between group1 (control group) and group2 (treated with DCs without being pulsed with tumor lysate). Average surviving time of group3 and group4 mice(treated with 5×105 and 5×104 tumor lysate-pulsed DCs respectively) was obviously longer than that of group1 and group2 (P<0.01), meanwhile the average surviving time of group3 was longer than that of group4(P<0.05). Tumor growth velocity of group3 and 4 was slower than that of group1 and 2(P<0.01), moreover the growth velocity of group3 was slower than that of group4(P<0.05). The average weight of bearing tumor mice decreased gradually, and that of group1 and 2 was obvious lower than that of group3 and 4 (P<0.05). In anaphase, the body weight of group3 mice was lighter than that of group4 (P<0.05). The level of the serum IL-6 of bearing tumor mice was higher than that of contral group mice(P<0.01), furthermore the level of the IL-6 of the mice treated with tumor lysate-pulsed DCs was highest(P<0.01) in three groups. The level of TNF-α of bearing3. tumor group was lower than that of control group(P<0.01), and the TNF-α level of treated group was obviously higher than that of control group and bearing tumor group(P<0.01). 4. In immunopreventive research, there was no tumor growing in group B and group C injected s.c with tumor lysate-pulsed DCs 1×105 and 1×104 respectively before inoculated with 5×105 B16 cells. A month later, these mice were inoculated with 5×105 B16 cells again, no tumor growed in group B still, but in group C there was five mice detected melanoma growth, but average surviving time of the bearing tumor mice was obviously longer than that of group A injected tumor cells only(P<0.01).Conclusions: The bearing tumor rate of mice is correlated with the number of the tumor cells, the larger quantity of the tumor cells is, the higher bearing tumor rate and the shorter average surviving time of the mice are. Tumor lysate-pulsed DCs has immunotherapy effect, but mature insensitive DCs have no immunotherapy effect. Immunotherapy effect is correlated to the quantity of tumor lysate-pulsed dendritic cells in a certain range. The therapy effect of 5×105 cells is better than that of 5×104 cells. The levels of IL-6 and TNF-α bearing tumor mice in vivo increased obviously after immunotherapy of tumor lysate-pulsed DCs, which suggests that tumor lysate-pulsed DCs enhance anti-tumor immunity. Tumor lysate-pulsed dendritic cells have immunoprotective effect to tumor bearing mice.
|
PDF Full Text Request