Inducing maturation of monocyte-derived dendritic cells on human epithelial cell feeder layer

ABSTRACT

Nowadays, dendritic cells [DCs] have a special place in cancer treatment strategies and they have been used for tumor immunotherapy as they can induce immune response against tumor cells. Researchers have been trying to generate efficient dendritic cells in vitro; therefore, this research was done to generate them for use in research and tumor immunotherapy. This study took place at Urmia University in 2010-2011 years. In this study plastic adherent monocytes were incubated with granulocyte-macrophage colony stimulating factor [GM-CSF] and interleukin-4 [IL-4] for five days. Finally, fully matured and stable DCs were generated by 48 hours of incubation in a monocyte conditioned medium [MCM] containing tumor necrosis factor-alpha [TNF-alpha] and epithelial cells. Phenotypic and functional analysis were carried out by using anti-CD 14, anti-CD80, anti-CD86, and anti-CD83 monoclonal antibodies, and by determining their phagocytic activity, mixed lymphocyte reaction [MLR] and cytokine production, respectively. Dendritic cells were produced with high levels of surface molecule, i.e. of CD80, CD83, CD86, HLA-DR, expression and low levels of CD14 expression. Dendritic cells showed efficient phagocytosis and ability to stimulate T-lymphocytes. Moreover, dendritic cells could secrete high levels of interleukin-12 [IL-12] cytokine which was depictive of their full maturation. Measurement of the produced cytokines showed the generation of type-1 dendritic cells [DC1]. Our study showed that skin epithelial cells could induce maturation of monocyte-derived dendritic cells [DCs]. This feeder layer led to the production of efficient dendritic cells with the ability to be used for tumor immunotherapy