Amplification and Identification of Immature Dendritic Cells Resistant to Maturation from Human Peripheral Blood Monocytes / 中国医师杂志

ABSTRACT

Objective To establish a method for in vitro induction and amplification of immature dendritic cells(DCs) with maturation resistance from human peripheral blood. Methods Mononuclear cells separated from peripheral blood were cultured with rhGM-CSF and rhIL-4 for 9 days, and rhIL-10 was added into medium at the 7th day. The suspending cells were examined with scanning electronic microscope and flow cytometry, and their ability for stimulating non-sensitized T lymphocyte proliferation was observed by mixed lymphocyte reaction(MLR). Cultured cells were stimulated with LPS and TNF-? for additional 2 days, respectively, and MLR was performed again. Results rhGM-CSF+rhIL-4-induced and IL-10-treated peripheral blood mononuclear cells(PBMC) exhibited typical morphological characteristics and immunological phenotype of DCs with high expression of CD1a and no expression of CD 83 on the cellular surface. Costimulating molecules CD 40 and CD 86 expressions were down-regulated.The capability of cultured cells for stimulating the proliferation of non-sensitized T lymphocyte was weak, and the same result was observed in cultured cells stimulated with LPS or TNF-?. Conclusion Immature dendritic cells with maturation resistance were obtained by culturing with IL-10,which might be a useful in the induction of immune tolerance of allogenic skin grafting for the major burn patients with deep burn wounds.