[In vitro experimental study on individualised immunotherapy induced by dendritic cells transfected with total RNA of autologous gastric cancer cells].
OBJECTIVE
To explore the efficiency of antitumor immunity induced by autologous dendritic cells (DCs) transfected with total RNA of autologous gastric cancer cells.


METHODS
Short-term cultured primary gastric cancer cells were prepared. DCs in peripheral blood mononuclear cells (PBMCs) from gastric cancer patients were induced with rhGM-CSF, rhIL-4 and TNF-alpha. The mature DCs transfected with total RNA of autologous gastric cancer cells were subjected to activate autologous T cells transforming into CTLs, and the activity of CTLs was detected by using CCK-8 kit. The immunological function of DCs were evaluated by flow cytometry and mixed lymphocyte culture(MLC) assay. The levels of IFN-gamma and IL-12 were detected by ELISA.


RESULTS
Mature DCs transfected with total RNA of autologous gastric cancer cells not only highly expressed costimulatory molecules (CD80, CD83 and CD86) and (MHC-I and MHC-II), but also powerfully stimulated allogenic or autologous T cell proliferation. The level of IL-12 secreted by mature DCs transfered with tumor RNA was notably higher than those secreted by untransfered and immature DCs, and the rate of killing autologous gastric cancer cells by CTLs was markedly higher than that of killing allogenic tumor cells.


CONCLUSIONS
Mature DCs transfected with autologous gastric cancer cell total RNA can induce and activate high antigen-specific CTLs directed at autologous gastric cancer cells in vitro.
Xie
Shao-jian
SJ
Department of Surgery, Surgical Research Center, Second Hospital Affiliated to Hebei Medical University, Shijiazhuang 050000, China. xshaojian@163.ccom
Yan
Qing-hui
QH
Shan
Bao-en
BE
Fu
Ze-xian
ZX
Meng
Fan-jie
FJ
Li
Bao-dong
BD
Cai
Jian-hui
JH
D015703:Antigens, CD / Q000378:metabolism; D018122:B7-1 Antigen / Q000378:metabolism; D051940:B7-2 Antigen / Q000378:metabolism; D003602:Cytotoxicity, Immunologic / Q000235:genetics / Q000276:immunology*; D003713:Dendritic Cells / Q000276:immunology* / Q000378:metabolism*; D006801:Humans; D007136:Immunoglobulins / Q000378:metabolism; D007167:Immunotherapy / Q000379:methods*; D018664:Interleukin-12 / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000378:metabolism; D008858:Microscopy, Phase-Contrast; D012313:RNA / Q000235:genetics*; D013274:Stomach Neoplasms / Q000235:genetics / Q000628:therapy*; D014407:Tumor Cells, Cultured; D000097802:CD83 Antigen
D015703:Antigens, CD; D018122:B7-1 Antigen; D051940:B7-2 Antigen; D007136:Immunoglobulins; D018664:Interleukin-12; D008562:Membrane Glycoproteins; D012313:RNA; D000097802:CD83 Antigen