The Research Of DC-CTL Immune Cell Targeting Therapy And Its Killing Effect On Ovarian Cancer

Objective This study investigated the anti-tumor effect of cytotoxicity T lymphocytes(CTL)activated,and we targeted at ovarian serous papillary cystadenocarcinoma in the nude mice.We investigated different injection frequency of DC-CTL to inhibit ovarian cancer in nude mice,and the effects on the expression of HMGA2 was provided the theoretical basis for the DC-CTL test used in clinical treatment,in order to advance ovarian cancer clinical trials.Methods(1)After isolated from umbilical cord blood by a high-gradient magnetic cell sorting system(MACS),CD34+ cells were cultured to differentiate into DC induced by a cocktail cytokiunes.Soluble antigen generated from SKOV3 ovarian carcinoma cells by freeze-thaw method was loaded on DC of DC culture.The expression of DC phenotypes such as CD80,CD83,CD86 and HLA-DR were examined by flow cytometry.DC function was evaluated by theirability to induce proliferation of allogeneic T cells in mixed lymphocyte reaction(MLR)assay.MTT assay were used to detect antigen-loaded CTL,CTL and monocytes T cell killing activity of SKOV3 cells.(2)We have established the nude mice xenograft model of ovarian cancer,and then followed by a serial number in each group of mice in the first 21 days on the right inoculated subcutaneously 0.2 m L dendritic cell vaccine in the first 24 and 27 days were injected 0.2ml saline as placebo;No.2 for the mice in the first 21,24 twice a day to do the treatment,in the first 27 days placebo;No.3 for the mice in the first 21,24,27 three times a day to do the above treatment;number of nude 4 days and 3 in the first 21,24,27 0.2ml normal saline injection.Observed growth in nude mice in each group were measured their body weight,tumor volume and weight;detect the expression of tumor in each group HMGA2 protein using immunohistochemical staining.Results(1)After freezing and thawing ovarian cancer cell line DC morphology antigen stimulation showed a mature state,and can express a variety of higher DC differentiation-associated antigens(CD80,CD83,CD86 and HLA-DR),while a stronger stimulus allogeneic T lymphocyte proliferation.MTT assay results showed that CTL antigen load on SKOV3 cell killing activity was significantly higher than CTL and monocytes T cells.(2)Ovarian cancer xenografts in nude mice were not accidental death case,only see a slight slow activity,in addition to no other obvious symptoms.Each DC-CTL weight of nude mice injected group,tumor volume and weight than the control group,which received 3 injections DC-CTL nude body weight,tumor volume and weight were significantly lower than the rest of the group.Immunohistochemistry results showed that the expression of DC-CTL injection group nude HMGA2 protein in tumor tissues was significantly lower than the control group,in which the expression of HMGA2 three times DC-CTL injected in nude mice was significantly lower than the rest of the organization each group.Conclusions(1)CTL activated by DC loaded with freeze-thaw ovarian carcinoma antigen has a higher proliferative ability and effective cytotoxicity against ovarian carcinoma cells in vitro.(2)DC-CTL could inhibit tumor growth in ovarian cancer,which may be related to inhibit the expression of HMGA2 protein.