| Background:Kidney cancer accounts for 3% of the Systemic malignant tumor. The incidence of kidney cancer second to yhe incidence of bladder tumor.It is not sensitive to radiation and chemotherapy. The patients of limited kidney cancer can be obtained by surgical treatment healing. But about 30% of kidney cancer patients had been found get tumor when it metastasis. Recently, studies show that the immune cells CD8+T (CTLs) plays a vital role in antineoplastic immune, but the activation process of CTL needs antigen presenting cells(APC). Dendritic cells (DC) is the strongest antigen presenting cells, DC cells which was sensitied by autologous ruptured tumor cells activate cytokine induced killer cells(CIK)can play a role in immune.This research explore the the function of kidney cancer immune therapy by the study of the DC-CIK which were sensited by related antigen of kidney G250.Methods:1,Induction and amplification of cytokine induced killer cells (CIK)Take 10ml peripheral blood,diluted with equal amount of PBS and join 10ml lymphocyte separation medium. Collecting layer cell, cultivate in 37℃,5% CO2 incubator. Collect the Non-adherent cells and join IFN-y,move to the flask which were coated by CD3MeAb10μg/ml×5ml. Inducte CIK cells by CIK broth.2,Induction and amplification of dendritie cell(DC)Take 10ml peripheral blood,diluted with equal amount of PBS.Adding 10ml Ficoll under the blood at the bottom of centrifugal pipe. Collect lymph cells row layer cells, cultivate in 37℃,5% CO2 incubator. Collect the adherent cells and add 10% RPM1164. Half quantitative change fluid develop.Get DC cells after 8 days.3,DC-CIK,DC-CIK-G25 in vitroCollect suspension cells which were cultivated by mature DC and CIK with CIK medium. Mixed the cells at the ratio of DC:CIK=1:4 after cells count.Get DC-CIK after cultivated by CIK medium.Join related antigen in kidney G250 to the mature DC in 750ug/L. cultivate in 37℃,5% CO2 incubator. Collect suspension cells which were cultivated by mature DC and CIK with CIK medium. Mixed the cells at the ratio of DC:CIK= 1:4 after cells count.Get DC-CIK after cultivated by CIK medium.4,Experimental group and observing indexesExperimental group for:blank group (A), (B), target control group (C), effect of experimental group (D). Each set three after hole respectively. Each group all respectively make 4 points in time, processing 1,3,5,7 days, Record the IL-2,IL-12,IFN-yexpression by RT-PCR.Record cellular phenotype analysis of all cells after 7 days. And study the killing effect of each groups against kidney cancer cells.Findings:Culturing DC and CIK 7 days,the expression of CD86,CD83 in DC increases. Expression ratio of CD83 is 80.3% and 86.4% in CD86. Expression ratio of CD 16 and CD56 is 47.4% in CIK.Over time, IL - 2 express in each groups increasing gradually in addition to control group.The IL - 12 express also increasing gradually as IL-2. There is no significant difference between each groups.On the 1,3,5 day,the IL-12 and IFN-y express higher than other groups. There is no significant difference between each groups.With the T ratio increasing,the killing effect of CIK against ACHN is enhancing.The effect when T ratio at 20:1 higher than the effect when T ratio at 5:1 (F=2569.29,q=154.47,P<0.01).The killing effect of DC-CIK group higher than CIK group at the same T ratio (F=2731.15～6982.21,q=35.284～415.3,P<0.01). The killing rate of DC-C1K-G250 group significantly higher than CIK group and DC-CIK group,it reached as high as 90.2% when T ratio at 20 : 1.Interpreation:Kidney cancer cells related antigen G250 sensitization DC co-cultivated with CIK can improve the immune function for kidney cancer,it aiso can restrain kidney cancer cells. Immune treatment in combination with drugs targeted therapy as a kind of new method of biological treatment for kidney cancer. The new method of biological treatment for kidney laid the experimental basis for clinical cancer treatment,it used for clinical cancer treatment have good prospects. |
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