Study Of Dendritic Cells Influence On CIK Cells Specific Cytotoxicity Against Ovarian Carcinoma Cells

Objectives: A. To compare the quantity, immunophenotype, andfunction of CD34+ stem cells and monocytes origin DCs. B. To study the effect of CD34+ stem cell DCs transfected with ovarian carcinoma total RNA on in vitro cytotoxicity of CIK cells.Methods: A. CD34+ stem cells were isolated from cord blood byimmunomagnetic bead assay, and monocytes were purified by plastic adherence. B. SCF, FL, GM-CSF, IL-4, and TNF-a were added to the culture system of stem cells to induced DCs; GM-CSF, IL-4, and TNF-a were added to the culture system of monocytes for the same purpose. C. Expansion and induction of stem cells to DCs was calculated by cell counting cell. D. Flow cytometer was used to explore the immunophenotype of different origin DCs. Function of DCs was observed by mixed lymphocyte reaction. E. Trizol harvested total RNA from ovarian carcinoma cell line SKOV3 and H08910. The RNAs were loaded to stem cell DCs as tumor antigens. F. T-INF, IL-1B , OKT-3, and IL-2 were added to cord blood mononuclear cells culture system to induce CIK cells. Expansion of CIK cells were calculated by cell counting every 3 days. Immunophenotype was explored by flow cytometer. G. Effectors were grouped accordingly as CIK cells co-cultured with DCs transfected with SKOV3 RNA, CIK cells co-culturedwith DCs transfected with H08910 RNA, CIK cells co-cultured with unloaded DCs. and CIK cells. Targets were SKOV3 and H08910 cell lines. In vitro cytotoxicity was examined by LDH release assay.Results: A. 0.78 + 0.31% of CBMC can be purified by Mini-MACSas CD34+ stem cells. B. The number of CD34+ stem cells can expand to 40. 24 + 9. 86 fold after 14 days. C. No matter in the expression of CD1a, CD80, CD86, and HLA-DR, or in the function of stimulating xenogenous lymphocyte proliferation, there was no difference between CD34+ stem cell DCs or monocyte DCs. D. The percentage of CD3+CD56+ cells is the same in CIK cells co-culture with DCs transfected with SK0V3 RNA, CIK cells co-culture with DCs, and CIK cells. E. The expansion rate of CIK cells can be accelerated by co-culturing with loaded or unloaded DCs. However, the expansion rate between loaded or unloaded group is the same. F. The strongest cytotoxicity against SK0V3 cell line was achieved by CIK cells co-cultured with DCs loaded with SKOV3 RNA. Cytotoxicities of CIK cells co-cultured with DCs loaded with H08910 RNA and CIK cells co-cultured with DCs are lower accordingly. CIK cells group possess the lowest tumor lytic ability among the 4 groups. Finding was similar when the target changed to H08910 cell line.Conclusions : A. CIK cells can be induced and expanded from CBMC.CIK cells are highly efficiency effectors for immunotherapy of ovarian carcinoma. B. CD34+ stem cells are suitable DC progenitors becausegreat amount of DCs can be induced and expanded from them. With loading of Trizol extracted total RNA of tumor cells, DCs can be used to arose powerful tumor cell lytic ability of CIK cells.