Effects of transfecting immature dendritic cells with indoleamine 2,3-dioxygenase gene on the spleen CD4~+ T cells in ovalbumin-allergized mouse model / 解放军医学杂志

ABSTRACT

Objective To observe the effects of mouse indoleamine 2,3-dioxygenase (IDO) gene on CD4+ T cells by transfecting an eukaryotic expression vector containing mouse IDO gene fused with enhanced green fluorescent protein (pEGFP-N1). Methods The immature dendritic cells (imDCs) derived from mouse bone marrow were cultured in vitro and morphologically identified by transmission electron microscope, scanning electron microscope and flow cytometry. The immature dendritic cells were transfected by DOTAP liposome, and the expression of green fluorescence protein was confirmed with inverted fluorescence microscope. The spleen-derived CD4+ T lymphocytes from mice were isolated and purified in vitro, and then carried to mixed lymphocyte reaction (MLR) with 3H-TdR incorporation. The effect of IDO gene on the proliferation of spleen-derived CD4+ T lymphocytes of the allergized mouse challenged by ovalbumin was assayed by 3H-TdR MLR incorporation, and the apoptosis of spleen-derived CD4+ T lymphocytes was assayed by TUNEL. Results The proliferation of spleen-derived CD4+ T lymphocytes of the allergized mouse challenged by ovalbumin were inhibited by the imDCs transfected with IDO gene, the number of CD4+ T lymphocytes treated with the imDCs transfected pEGFP-N1-IDO eukaryotic expression vector decreased significantly compared with that in control group (P