earticle

영어

It is well known that the antigenic vaccines shows various side effects and systhetic peptide humoral and cellular immune responses. To overcome these problems, we developed the effective antigen delivery system like antigen-antibody complex of EpCAM-Fc or EpCAM-Fc KDEL to dendritic cells (DCs) which are a representative antigen-presenting cells to T cells. First, conventional Dcs were generated by using human umbilical cord blood which is a rich source of hematopoitic stem cells, progenitor cells and immune cells including dendritic cells (DCs)at an immature stage of differentiation. When conventional DCs were stimulated with LPS (1 ㎍, 48 hours), the expression level of CD80, CD 86 and MHC Ⅱon the cellular surface was increased, confirming the molecular and cellular properties of the activated conventional DCs. Next, the antigen-antibody complexes of EpCAM-Fc (insect-EpCAM-Fc) or EpCAM-Fc KDEL (insect-EpCAM-Fc KDEL) were expressed in heterologous expression system of insect cells such as Sf9 and Hi5 cel lines in which glycosylation of glycoprotein is humanized and followed by anatlyzing their glycosylation patters and in vitro biological activities. When insect EpCAM-Fc or EpCaM-Fc KDEL (10 ㎍, 48 hours) were treated to conventional DCs, conventional DCs were not activated at all. However, when insect EpCAM-Fc or EpCAM-Fc KDEL (10 ㎍, 48 hours) were treated to conventional DCs with low dose of LPS (0.01 ng), conventional DCs were activated well showing the increased expression of CD80, CD 86 and MHC Ⅱ on their cellular surface. Taken together, the heterologous expression system using insect cells could be used to generate antigen-antibody complexes to activate DCs, providing an alternative application system to develop the ani-cancer vaccines.