The Cytotoxicity Of Cytokine-Induced Killer Cells Against Non Hodgkin Iymphoma Cell Line Precondition With Cisplatin

Objective:DDP (cisplatin) is wildly used in the treatment of non-Hodgkin’s lymphoma and other neoplasm, which is used as a second-line drug treatment of recurrent and refractory non-hodgkin’s lymphoma. However, many adverse reactions show up during treatment, and the curative effect is poor. With the development of the cell immunology and molecular biology, chemotherapy combined with immunotherapy has been applied in clinical. Cytokine-induced killer cells have advantages such as large proliferation; a wide variety of tumor cell killing activity; non-MHC restricted; no cytotoxic effect on endogenous tissue. Research suggests that pretreated tumor cells with small dose of cisplatin (DDP), can reduce the adverse reaction of DDP, and also enhance the cytotoxicity of CIK cells. This study is aim to analysis the cytotoxicity of cytokine-induced killer cells against non Hodgkin lymphoma cell line precondition with cisplatin.Methods:The logarithmic phase cells of human NHL cell lines Namalwa (Burkitt lymphoma cell lines) which cultured in vitro and SU-DHL-4(diffuse large B cell lymphoma cell lines) were collected for experiment. Ficoll density gradient centrifugation was used to obtain peripheral blood mononuclear cell. Interferon-y (1000u/ml) was added in the first day, after then placed in saturated humidity incubator (37℃,5%CO2) for24hours. The next added Interleukin-1(IL-1)100u/ml, Interleukin-2(IL-2)500u/ml,CD3McAb50ng/ml and cultured in saturated humidity incubator (37℃,5%CO2). Fresh culture medium were replaced every3days, human recombinant interleukin-2was supplied as well. CFSE/PI double labeling method was used to detect the effect of CIK cells against Non Hodgkin lymphoma cells which were pretreated by DDP in different concentration (Ong/ml、25ng/ml、50ng/ml、75ng/ml) and different time (18h、24h、36h), and then compared by the cytotoxicity.Results:CIK cells which were induced in vitro were obtained after21days, with ideal ratio of CD3+CD56+and CD3+CD8+cells to be effectors cells. The target cells were pretreated by four concentrations of DDP, with the increased of DDP concentration, the cytotoxicity of CIK cell kill lymphoma cells increased. The differences were statistically significant between the two groups with and without CIK cells (P<0.05). The target cells were continued to culture after pretreated by DDP. Different culture time cells were selected and added CIK cells. We found that with the DDP pretreatment time extended the cytotoxicity of CIK cells kill lymphoma cells enhanced. The difference were statistically significant between the two groups with and without CIK cells (P <0.05).Conclusion:The cytotoxicity of CIK cell kill lymphoma cells was increased after lymphoma cells pretreated by DDP. The combined effects of DDP and immune therapy have a synergistic anti-tumor effect.