Genetically Manipulated Human Embryonic Stem Cell-derived Dendritic Cells with Immune Regulatory Function

¹ The Department of Immunogenetics, Graduate School of Medical Sciences, Kumamoto University, Kumamoto, Japan
² Laboratory of Embryonic Stem Cell Research, Stem Cell Research Center, Institute for Frontier Medical Sciences, Kyoto University, Kyoto, Japan
³ Laboratory of Molecular Medicine, Human Genome Center, Institute of Medical Science, University of Tokyo, Tokyo, Japan
⁴ Department of Development and Differentiation, Institute for Frontier Medical Sciences, Kyoto University, Kyoto, Japan

^* To whom correspondence should be addressed. E-mail: senjusat{at}gpo.kumamoto-u.ac.jp.

	Abstract

Genetically manipulated dendritic cells (DC) are considered to be a promising means for antigen-specific immune therapy. This study reports generation, characterization, and genetic modification of dendritic cells (DC) derived from human embryonic stem (ES) cells. The human ES cell-derived DC (ES-DC) expressed surface molecules typically expressed by DC, and had capacities to stimulate allogeneic T lymphocytes and to process and present protein antigen in the context of HLA class II molecule. Genetic modification of human ES-DC can be accomplished without the use of viral vectors, by the introduction of expression vector plasmids into undifferentiated ES cells by electroporation and subsequent induction of differentiation of the transfectant ES cell clones to ES-DC. ES-DC introduced with invariant chain-based antigen-presenting vectors by this procedure stimulated HLA-DR-restricted antigen-specific T cells in the absence of exogenous antigen. Forced expression of PD-L1 in ES-DC resulted in the reduction of the proliferative response of allogeneic T cells co-cultured with the ES-DC. Generation and genetic modification of ES-DC from non-human primate, cynomolgus monkey, ES cells was also achieved by the currently established method. ES-DC technology is therefore considered to be a novel means for immune therapy.