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Impact of transfection with total RNA of K562 cells upon antigen presenting, maturation, and function of human dendritic cells from peripheral blood mononuclear cells
Author(s) -
Gao Li,
Fan Huahua,
Lu Huazhong,
Nie Xiaoxuan,
Liu Yan,
Yang Yiming,
Qian Kaicheng,
Gao Feng
Publication year - 2007
Publication title -
transfusion
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.045
H-Index - 132
eISSN - 1537-2995
pISSN - 0041-1132
DOI - 10.1111/j.1537-2995.2007.01098.x
Subject(s) - ctl* , cytotoxic t cell , transfection , k562 cells , microbiology and biotechnology , biology , dendritic cell , antigen , peripheral blood mononuclear cell , immune system , cd80 , immunology , cell culture , cd40 , cd8 , in vitro , leukemia , biochemistry , genetics
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). STUDY DESIGN AND 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. CONCLUSION: 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.