Adenoviral transduction of human ‘clinical grade’ immature dendritic cells enhances costimulatory molecule expression and T-cell stimulatory capacity
- 作者：Rouard ; Hé ; lè ; ne ; Lé ; on ; Anne ; Klonjkowski ; Bernard ; Marquet ; Jeanine ; et. al.
- 关键词：Human dendritic cell-Anti-tumor vaccination-Adenoviral transduction ; PBMC ; peripheral blood mononuclear cell ; PBL ; peripheral blood lymphocyte ; DC ; dendritic cell ; Mo ; monocyte ; CTL ; cytotoxic T lymphocyte ; S.D. ; standard deviation
- 刊名：Journal of Immunological Methods
- 出版年：2000
- 期刊代码：57_00221759
- 类别：imm
- 出版时间：July 31, 2000
- 卷：241
- 期：1-2
- 页码：69-81
- 文件大小：1.01 M
摘要
The therapeutic use of dendritic cells (DC) in antigen-specific anti-tumor vaccines, requires sufficient numbers of functional DC, the preparation of which should comply with the code of Good Manufacturing Practice. In addition, the expression of tumor specific antigen should be possible in these DC. As a preclinical step, the method reported here was developed in healthy volunteers. Monocytes (Mo) were isolated by leukapheresis from 12 donors, purified by elutriation and then cultured for 6 days in sealed bags in AIM-V serum free medium with granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-13 (IL-13). Between 6×108 and 1×109 immature DC (iDC) could be differentiated from one leukapheresis. Cells displayed a characteristic iDC phenotype (CD1a+, CD14−, CD80+, CD86+, HLA DR+, CD83−), and had potent allogeneic and antigen dependent autologous T cell-stimulatory capacity. Moreover, iDC could be further differentiated into mature DC by CD40 ligation as assessed by CD83 expression and the upregulation of HLA-DR and costimulatory molecules. After infection with a recombinant adenovirus encoding for beta-galactosidase (βGal), 50%to 80%of iDC expressed βGal without toxicity. Adenovirus infection increased the expression of both costimulatory molecules and CD83, and also increased allogeneic stimulatory capacity. Thus, the method developed here allows us to use large numbers of functional iDC as will be required for therapeutic uses in man. These DC can express a transgenic protein.