Abstract:
: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 6x10(8) and 1x10(9) 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 (betaGal), 50% to 80% of iDC expressed betaGal 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.
journal_name
J Immunol Methodsjournal_title
Journal of immunological methodsauthors
Rouard H,Léon A,Klonjkowski B,Marquet J,Tennezé L,Plonquet A,Agrawal SG,Abastado JP,Eloit M,Farcet JP,Delfau-Larue MHdoi
10.1016/s0022-1759(00)00214-3keywords:
subject
Has Abstractpub_date
2000-07-31 00:00:00pages
69-81issue
1-2eissn
0022-1759issn
1872-7905pii
S0022-1759(00)00214-3journal_volume
241pub_type
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pub_type: 评论,杂志文章
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journal_title:Journal of immunological methods
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