Subtypes And Immune Function Of Vγ9Vδ2T Cells In The Peripheral Blood Of Hcc Patients And Healthy People When Before And After Amplification In Vitro

Objective:To investigete the difference of percentage, mature classification and Immune killing function of Vγ9Vδ2T cell in peripheral blood between patients with HCC and healthy person, the amplification effect and index changes after amplification in vitro.Methods:10 ml were extracted from the peripheral blood of HCC patients(n = 25) and healthy donors(n = 20), Peripheral blood mononuclear cells(PBMCs) were isolated by the discontinuous density gradient centrifugation.1. Proportion, mature and differentiate subtypes and IFN-γ and CD107 a expressing of the Vγ9Vδ2T cells were detect by using flow cytometry.2. The separation of lymphocytes were joined into GT- T551 lymphocyte culture medium with oledronate and human IL – 2, which were placed at 37 ℃, 5%CO2incubator. regularly join human IL – 2 every 2-3 d, we harvest cells after 12-14 d.3. Proportion, mature and differentiate subtypes and IFN-γ and CD107 a expressing of the Vγ9Vδ2T cells were detect by using flow cytometry again.Results:1. In healthy group 18 cases were trainned successfully, 2 cases were failure in amplification. In HCC group 21 cases were trainned successfully, 4 cases were failure in amplification.2. The percentage of Vγ9Vδ2Tcell of total T cell in peripheral blood of HCC patients is lower than healthy people in front of the train. The differentiation of mature proportion 、 proportion of secreting IFN- γ and expressing CD107 a all have no difference comparing with healthy people in front of the train.3. The proportion of Vγ9Vδ2T cell in peripheral blood of HCC patients increased significantly after augmentation in vitro. When matured subtypes of Vγ9Vδ2T cell were compared before and after culturing in vitro, after training the proportion of Tn,Tcm and Temra of HCC patients and healthy group were significantly lower than before the training, proportion of Tem, Tem+Temra of HCC patients and healthy group were higher than after the training. Proportion of secreting IFN- γ and expressing CD107 a all have no difference between before and after amplification in HCC patients and healthy group.4. The percentage of Vγ9Vδ2T cell of total T cell,the mature proportion,proportion of secreting IFN- γ and expressing CD107 a all have no difference between HCC patients and healthy people after amplification.Conclusions:1. The percentage of Vγ9Vδ2T cell in peripheral blood of HCC patients is lower than healthy people in front of the train. The distribution of matured subtypes of HCC patients are similar to healthy people. Secreting IFN- γ and expressing CD107 a all have no difference with healthy people.2. Applying ZOL+ IL-2 when culturing PBMCs of HCC patients and healthy people in vitro can highly amplify Vγ9Vδ2T cells.Total effect type of matured subtypes of Vγ9Vδ2T cell of HCC patients and healthy people increased highly after augmentation in vitro. proportion of secreting IFN- γ and expressing CD107 a all have no difference between HCC patients and healthy people after amplification.3. This approach can be used to provide appropriate Vγ9Vδ2T cells for the next step of experiment researchs and clinical applications of adoptive cellular immunotherapy.