Stimulation Of γδT Cell In Peripheral Blood By Zoledronic Acid And Its Cytotoxic Effect In Nonspecific Immunity Of Hepatoma.

PartⅠγδT lymphocyte multiplication stimulated by zoledronic acidObjective To study the effect ofγδT lymphocyte multiplication stimulated by zoledronic acid .Methods: Take 10ml peripheral blood of healthy adults, obtain peripheral blood mononuclear cell by centrifugation on a Ficoll-Hypaque density gradient. Take zoledronic acid the Third-generation Biphosphonate as the stimulating factor.Exert different concentration zoledronic acid combined with IL-2 to stimulate peripheral bloodγδT cells proliferation. Collecting cells after 12-14 days, analyze correlated phaenotypes by flow cytometry. The quantity of cells was determined by method of 3H-TdR incorporation.Results: After cultured 12-14 days ,peripheral bloodγδT cells obviously multiplied. Confer with the control group, proportion ofγδT phaenotype in experimental gtoup obviously heightened(control group 9.7%,experimental gtoup 80.8%,P<0.01). proportion of CD3 phaenotype in experimental gtoup also obviously heightened(control group 71.2%,experimental gtoup 96.2%,P<0.01)。Cells in experimental gtoup obviously multiplied determined by method of 3H-TdR incorporation(CMP of control group 9961.5,CMP of experimental gtoup 17488.6,P<0.01).In all of the zoledronic acid concentration,γδT cells multiplied most obviously in 1μΜzoledronic acid.γδT cells.Conclusions: Peripheral bloodγδT lymphocyte can be stimulated and multiplied by zoledronic acid ,and can obtain quantity of highly purifiedγδTcells. PartⅡThe cytotoxic activity of the multipliedγδT lymphocyte to hepatoma carcinoma cell.Objective To investigate the cytotoxic activity ofγδT lymphocyte stimulated and multiplied by zoledronic acid to hepatoma carcinoma cell.Methods: Obtain peripheral bloodγδT cells of healthy adults by the method of experiment one .TakeγδT cell as effector cell,and hepatoma carcinoma cell(SMCC-7721,MHCC-97,Hep - G2) as target cell. Mixed culture such cells according to the efffector/target cell ratio(E/T 10:1,20:1).Stop experiment after 48 hours. Determine the cytotoxic activity of effector cells to target cells by method of SRB.Results:γδT cell display powerful cytotoxic activity to the three tumour cells. The percentage of cytotoxin reached 98.7% when E/T was 20:1 and reached 92.6% when E/T was 10:1. The cytotoxic activity ofγδT cell to different tumour cells has no significant deviation in same E/T(P>0.05). The cytotoxic activity ofγδT cell to same tumour cells has significant deviation in different E/T(P<0.05)。Conclusions: TheγδT lymphocyte stimulated and multiplied by zoledronic acid has significant cytotoxic activity to hepatoma carcinoma cell in vitro.