Transfection of dendritic cells (DCs) with the CIITA gene: increase in immunostimulatory activity of DCs

Abstract

Dendritic cells ( DCs ) are the major antigen -presenting cells. They are able to present tumor antigens to immunologic effector cells. MHC class II molecules on DC surfaces play an important role in priming effector cells against tumor cells and their antigens. The transactivator CIITA ( MHC class II transactivator ) is a non ± DNA -binding transactivator, which regulates the expression of MHC class II, HLA -DM, and invariant chain and behaves as a master controller of constitutive and inducible MHC class II gene activation. Here, we transfected DCs with the CIITA gene using a novel transfection technique. The vector system consisted of a plasmid bound to an adenovirus via poly -L -lysine, which is covalently bound to a UV -irradiated adenovirus. After transfection, expression of MHC class II on DCs increased from 27% to 75% on day 2 after transfection. Transfected DCs were co -cultured with immunologic effector cells. Cytotoxicity of effector cells against tumor cells increased after co -culture with transfected DCs to 63% compared to 15% with effector cells co -cultured with irrelevantly transfected DCs ( P = .037 ) . This effect was dependent on the timing and period of coculture. In conclusion, transfection of DCs led to an increase in antitumoral immunostimulatory capacity of DCs. We can further conclude that DCs could be efficiently transfected with the CIITA gene. Transfection of DCs led to an increase in antitumoral immunostimulatory capacity of DCs and may have a major impact on immunotherapeutic protocols for patients with cancer.

Human cytokine-induced killer cells have enhanced in vitro cytolytic activity via non-viral interleukin-2 gene transfer

“…Flow cytometry showed CD14 negative populations, expressing markers typical for DC (CD80 + , CD83 + , CD86 + and HLA-DR expressing cells). The CIK cells were phenotyped with antibodies against CD3, CD8, CD16, CD40L, CD56, HLA-ABC and HLA-DR. Data was similar to other studies by our group (data not shown) [5,11,14]. …”

Generation of dendritic cell-based vaccines for cancer therapy

“…With the aid of liposomemediated infection, gene transfer into CD83 + DC resulted in more than 90% of the cells transduced. Using a protocol with UV-irradiated adenoviruses similar results can be obtained without addition of liposomes (Mulders et al, 1998;Märten et al, 2001). Adenoviral vectors can also be used for transduction of CD34 + cell derived DC .…”

Autologous Tumor Lysate-Pulsed Dendritic Cell Immunotherapy with Cytokine-Induced Killer Cells Improves Survival in Gastric and Colorectal Cancer Patients

“…Studies demonstrated that IFN-γ and IL-12 play critical roles in immunotherapy using DC and CIK [28], [33]. Tumor cells are highly heterogeneous [46] and a specific tumor may contain cells with both high and low MHC-I populations [47], [48].…”