Killing Effect Of CTLs Induced By Dendritic Cells After Loaded With Soluble Antigens Of Nasopharyngeal Carcinoma CNE Cells In Vitro

ObjectiveDentritic cells are important antigen presenting cells in human being. They can specially presenting antigen to T lymphocytes and initiate specific antitumor immunity reaction in vitro and in vivo. Their remarkable effectiveness is in large part due to their efficiency in capturing, processing and presenting antigens. Therefore, dentritic cells have become one of the hot spots in the field of tumor immuno-adoptive therapy since 90s.In this study, we assess the killing effect of CTLs after activated by DCs loaded with soluble antigens of nasopharyngeal carcinoma CNE cells in vitro MethodsMonocytes were isolated from normal human peripheral blood mononuclear cells(PBMCs), cultured with cytokines including GM-CSF, IL-4 and TNF- a to induce and expand mature DCs, which were observed and photographed by inversion microscope and electron microscope. Before and after loaded with soluble antigens of CNE cells, the phenotype of mature DC was detected and the function of DC tostimulate allogeneic cells proliferation was measured. Host lymphocytes were stimulated with DCs loaded with soluble antigen of CNE cells in the culture medium containing interleukin-2 (IL-2) for 7 days. The special lysis activity of CTLs against nasopharyngeal carcinoma CNE cells was analysed. Results1. The DCs derived from monocytes cultured in GM-CSF, IL-4 and TNF- a havest on day 7. The special configuration of DCs was observed and photographed by inversion microscope and electron microscope. Surface marker assayed by FACS showed that DCs expressed CD la, CD80, CD86, HLA-DR, CD83 and CD40 highly.2. After being loaded with soluble antigens of CNE cells, no significant changes in the surface markers and the cytomorphology of mature DCs were detected and mature DCs still have strong potential to stimulate the proliferation of allogenetic T cells.3. DCs loaded with soluble antigens of CNE cells were cultured with autologous PBMCs at the ratio 1:20,add IL-2 and half-renewed media every 3 days. At the 8th days, CTL cytotoxicity assays were performed by LDH kits. The results suggested that the CTLs have a special killing effect on nasopharyngeal carcinoma cells in vitro.ConclusionWe could obtain DCs , which have immunoactivity, with appropriate...