Adeno-associated virus-mediated Her2/neu gene transfection enhances the immunostimulatory capacity of human dendritic cells in vitro.
- Author:
Lei XU
1
;
Rong-cheng LUO
;
Zhong-hong BAI
;
Ruan-cheng XUE
;
Fang-ju WANG
Author Information
- Publication Type:Journal Article
- MeSH: Breast Neoplasms; pathology; Cell Line, Tumor; Cells, Cultured; Dendritic Cells; cytology; immunology; metabolism; Dependovirus; genetics; metabolism; Genes, erbB-2; genetics; Genetic Vectors; Granulocyte-Macrophage Colony-Stimulating Factor; pharmacology; Humans; Leukocytes, Mononuclear; cytology; Receptor, ErbB-2; biosynthesis; genetics; Recombinant Proteins; genetics; immunology; metabolism; Transfection
- From: Journal of Southern Medical University 2009;29(9):1893-1895
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo evaluate the immunostimulatory capacity of human peripheral blood dendritic cells (DCs) with Her2/neu gene transfection mediated by adeno-associated virus.
METHODSThe HLA genotypes of the breast cancer cells SK-BR-3 and MCF7 were determined, and the mononuclear cells from healthy donors with matching HLA genotype were isolated by Ficoll-Hypaque density gradient separation. The isolated cells were divided into two groups with or without transfection with the recombinant virus harboring Her2/neu gene. The cells were cultured for 7 days in RPMI 1640 medium supplemented with 10% AB human serum, GM-CSF, interleukin-4 (IL-4) and tumor necrosis factor-alpha (TNF-alpha). The mature DCs were then harvested from the cell culture and their phenotypes were identified using flow cytometry. MTT assay was employed to examine the specific killing activity of the T cells induced by the DCs.
RESULTSThe DCs transfected with the recombinant adeno-associated virus expressed CD1a, CD86 and CD83 at the rate of 98.10%, 99.42%, and 84.59%, and those without the viral transfection expressed the markers at the rate 92.69%, 98.07%, and 82.72%, respectively, showing no obvious differences in the phenotypes of the two DCs. The transfected DCs, however, showed markedly higher expression rates of CD40 and CD80 than the non-transfected DCs (61.02% vs 36.19%, and 97.61% vs 55.5%, respectively). The DCs, irrespective of the transfection, showed comparable capacities in stimulating T cell proliferation. The transfected DCs exhibited the capacity of inducing the T cells to specifically kill the target tumor cells, with the highest killing rate of (39.7-/+7.2)%.
CONCLUSIONThe immunostimulatory capacity of human peripheral blood DCs are enhanced by Her2/neu gene transfection mediated by adeno-associated virus.