Impact of transfection with total RNA of K562 cells upon antigen presenting, maturation, and function of human dendritic cells from peripheral blood mononuclear cells.
BACKGROUND
Vaccination of dendritic cells (DCs) with tumor antigens is a potential strategy to induce tumor-specific immunity in tumor-bearing patients. The purpose of this study was to investigate whether human monocyte-derived DCs were able to present P210(Bcr-Ab1) protein and induce antigen-specific cytotoxic T lymphocyte (CTL) responses in vitro after transfected with total RNA of K562 cells (K562-RNA).


METHODS
DCs derived from human peripheral blood mononuclear cells were transfected with K562-RNA with electroporation or DOTAP lipofection. The successful transfection was determined by reverse transcription-polymerase chain reaction and Western blot. The phenotypes of the DCs were analyzed by flow cytometry (FCM), and cytotoxicity of CTL was assessed by propidium iodide staining followed by FCM analysis. The CD1a expression and purity of DCs were measured by FCM.


RESULTS
The Bcr-Abl fusion gene was detected in the DCs with 24 hours after the transfection. The transfected cell expressed increased levels of CD80, CD83, CD86, and HLA-DR. Moreover, the transfected DCs strongly stimulated the T lymphocytes to gain cytotoxic activity against K562 cells. Culture medium containing 1 percent human plasma was the most effective for DC growth.


CONCLUSIONS
Human DCs transfected with K562-RNA effectively induce specific immune responses. This method can be used to induce tumor-specific immune response and may have potential application in immunotherapy of tumors.
Gao
Li
L
Blood Engineering Laboratory, Transfusion Institute of Shanghai Blood Center, Shanghai, China. corrie.gl@gmail.com
Fan
Hua-Hua
HH
Lu
Hua-Zhong
HZ
Nie
Xiao-Xuan
XX
Liu
Yan
Y
Yang
Yi-Ming
YM
Qian
Kai-Cheng
KC
Gao
Feng
F
D017951:Antigen Presentation / Q000276:immunology*; D019496:Cancer Vaccines / Q000235:genetics* / Q000276:immunology; D002454:Cell Differentiation / Q000276:immunology; D003713:Dendritic Cells / Q000166:cytology / Q000276:immunology*; D016044:Fusion Proteins, bcr-abl / Q000235:genetics; D006801:Humans; D016130:Immunophenotyping; D020014:K562 Cells; D009000:Monocytes / Q000166:cytology*; D012334:RNA, Neoplasm / Q000235:genetics* / Q000276:immunology; D014162:Transfection
D019496:Cancer Vaccines; D012334:RNA, Neoplasm; D016044:Fusion Proteins, bcr-abl