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Transfection of human monocyte‐derived dendritic cells with CpG oligonucleotides
Author(s) -
Erhardt Michael,
Gorschlüter Marcus,
Sager Jens,
Ziske Carsten,
Strehl John,
LilienfeldToal Marie V,
SchmidtWolf Ingo GH
Publication year - 2005
Publication title -
immunology and cell biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.999
H-Index - 104
eISSN - 1440-1711
pISSN - 0818-9641
DOI - 10.1111/j.1440-1711.2005.01328.x
Subject(s) - cpg oligodeoxynucleotide , transfection , cd80 , cd86 , electroporation , cpg site , cd40 , biology , microbiology and biotechnology , cd14 , cell culture , immunology , in vitro , flow cytometry , immune system , t cell , cytotoxic t cell , dna methylation , gene expression , gene , biochemistry , genetics
Monocyte‐derived dendritic cells (mDC), the most frequently applied DC subset in clinical studies, which can be obtained easily from peripheral blood monocytes after incubation with GM‐CSF and IL‐4, have not been clearly demonstrated to be activated by CpG oligodeoxynucleotides (ODN). The development of novel molecular strategies – such as the use of CpG‐ODN – to increase immunological functions and thus improve the therapeutic efficiency of mDC vaccines in the treatment of malignant diseases is highly desirable. CpG‐ODN need to be internalized into specific intracellular compartments to be active. Therefore, we applied electroporation and lipofection and compared these techniques with incubation to overcome possible defects in localization. Conditions of CpG‐ODN transfection of these cells were optimized using fluorescein‐marked ODN 2216. We were able to achieve high transfection efficiencies with various methods of delivery. However, we did not observe increased expression of maturation‐associated and functionally relevant surface antigens (CD14, HLA‐DR, CD40, CD83, CD80 and CD86), significant secretion of IL‐12 and IFN‐α in culture supernatant, or enhanced antitumour activation of cytokine‐induced killer cells. In conclusion, our results show that non‐viral transfection of CpG‐ODN is not sufficient to overcome resistance of mDC to CpG activation.