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

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.

Generation of Dendritic Cells from Human Peripheral Blood / 中国肿瘤生物治疗杂志

Dendritic cells (DC) are antigen presenting cells (APC) that play critical roles in the initiation of T cell response and development of T cell-dependent antibodies in vivo. CD34_+ hematopoietic progenitor cells of bone marrow and peripheral blood can differentiate into DC when cultured with GM-CSF and TNF-? in vitro. In the present study, we cultured monocytes isolated from human peripheral blood with 100ng/ml hGM-CSF and 500U/ml hIL-4 for one week, and then found that a large number of DC with high purity were gengerated. DC expressed MHC I , MHC II and costimuladng moleculers highly on cell surface and cound stimulate proliferation of allogeneic T lymphocytes. Self serum or fetal calf serum are best for generation of DC. When hGM-CSF was used alone, the monocytes differentiated into macrophages but not to DC. TNF-? could induce further maturation of DC when added in late period of the culture. Generation of DC from human peripheral blood may facilitate further studies on DC and their clinical applications.