Gene transfer of the CD40‐ligand to human dendritic cells induces NK‐mediated antitumor effects against human carcinoma cells

Abstract

The CD40-ligand (CD40L) is a key molecule for the activation of dendritic cells (DCs), followed by the induction of DC maturation and cytokine production. Here we found that DC infected with adenovirus vector encoding human CD40L (CD40L-DC) displayed significantly higher levels of immune accessory molecules and IL-12 production than did uninfected cells, and that CD40L-DC produced much higher levels of IFN-c. To investigate whether CD40L-DC-derived these soluble factors could stimulate NK cells without physical cell-to-cell contact, we cocultured NK cells with CD40L-DC in transwell culture plates. NK cells showed up-regulated cytotoxic activity toward various oral squamous cell carcinoma (OSC-70, HSC-2, HSC-3), and we determined that both IL-12 and IFN-c contributed to the CD40L-DC-mediated NK cell activation. NK cells stimulated with CD40L-DC resulted in the induction of the cell surface expression of TRAIL, the production of IFN-c and intracellular accumulation of granzyme B. The cytotoxic activity of NK cells stimulated with CD40L-DC could be mostly inhibited by neutralizing antibody for TRAIL and completely abrogated by the combination of antibody and exocytosis inhibitor, indicating that this was mainly mediated by a TRAIL-TRAIL-receptor interaction and granule exocytosis. Moreover, CD40L-DC-activated NK cells could induce up-regulation of a death-receptor TRAIL-R2 (DR5) and down-regulation of a decoy receptor TRAIL-R3 (DcR1) on carcinoma cells. Overall, these results have revealed that CD40L-DC could activate an innate immune reaction by stimulating NK cells followed by carcinoma cells, supporting that administration of CD40L-DC may have potential as an anticancer therapy. ' 2006 Wiley-Liss, Inc.Key words: CD40-ligand; adenoviral vector; dendritic cells; NK cells; IFN-g; TRAIL Dendritic cells (DC) are known to enhance the tumoricidal activity of natural killer (NK) cells, and it has been reported that murine DC activated murine NK cells to enhance cytotoxic activity and IFN-g production in vitro. 1 In that report, DC-induced NK cytotoxic activity and IFN-g secretion required DC-NK cell-tocell contact. However, the molecular mechanisms of the DC and NK cell interaction have not been fully clarified. It has also been reported that human DC induced human NK cytotoxic activity, which similarly required DC-NK cell-to-cell contact. Thus, IFN-a-or lipopolysaccharide (LPS)-pretreated human DC induced cytotoxic activity in cocultured human NK cells. 2-4 More recently, Borg et al. reported that DC-mediated NK cell activation required the formation of a synapse, which led to IL-12 polarization in DC. 5 In addition to the DC to NK cell contact-dependent mechanism, IL-12 or IFNs secreted from DC are also involved in initiating the activation of NK cells.It has been reported that precursor DC have the ability to produce type1 IFNs,6 and that precursor DC activated by viral infection are induced to increase the production of type1 IFNs, which activate NK cells. 7 CD40L is essential for the activation of DC, followed by t...

Gene Transfer of Noncleavable Cell Surface Mutants of Human CD154 Induces the Immune Response and Diminishes Systemic Inflammatory Reactions

“…In an attempt to investigate the biologic function of the newly constructed CD154 mutant, we next constructed modified adenovirus vectors driving CD154-WT or CD154-M3 genes with the CA promoter/enhancer and incorporating the integrin-binding motif, RGD, into the HI loop of the fiber knob (AxCACD154-WT and Ax3CACD154-M3). 51 At the same multiplicity of infection, expression of transgenes such as LacZ and green fluorescence protein after infection of CLL-B cells with fiber RGD-modified adenovirus vectors were approximately 10 times higher than that obtained with adenovirus vector containing wild-type fiber knob (data not shown). Using these fibermodified adenovirus vectors, we transfected CD154-WT or CD154-M3 mutant into CLL-B cells and found approximately equal levels of CD154 on the cell surface.…”

Dendritic Cells and their Receptors in Antitumor Immune Response

“…Using similar approach, CD40L transduced DCs enhanced the NK cell potential to kill various tumor cell lines. CD40L activated NK cells to express TRAIL and killed squamous cell carcinoma through TRAIL and granzyme-B [149]. DCs that were activated with CD40L transduced cell lines induced protective immune response against P815 mastocytoma upon loading with weakly immunogenic P1A tumor peptide of P815 mastocytoma in-vivo, which showed that CD40 induced efficient maturation of DCs [150].…”

Bidirectional Interactions of NK Cells and Dendritic Cells in Immunotherapy: Current and Future Perspective

“…Enhancing CD40L expression on DC cells, for example, enhanced NK-DC interactions and the maturation of DC, which in turn enhanced the production of appropriate cytokines that stimulated NK and enhanced the ability of these NK cells' cytotoxic ability toward certain tumor [43]. DC can also be activated by the innate recognitions of certain recombinant viruses or viral vectors (such as vesicular stomatitis virus, canarypox virus and adenoviral virus) to promote NK activation and prolonged antitumor immunity [44,45].…”