| Dendritic cells(DC) are the most important professional antigen-presenting cells (APC) in vivo, which play critical roles in innate and acquired immunity. They are heterogeneous group of cells that represent distinct origin, developmental stages and subsets. Dendritic cells differentiate from immature dendritic cells(imDC) to maure dendritic cells(mDC) after pathogens or alloantigens'stimulation. ImDC with deficient costimulatory molecules can induce T cell anergy and promote alloantigen-specific tolerance. In the treatment of severe burns, early and effective coverage of the wound is necessary. Allogeneic skin transplantation is a effective method, but the host versus graft reaction(HVGR) after transplantation leads to strong rejection within 3 weeks and limits its application. Prolongation of skin allografts survival by immunosuppressive drugs increases the potential risk for life-threatening infections, that is why clinical skin allografting practically does not exist. Therefore, it is crucial to induce host alloantigen-specific tolerance to donor.ImDC can effectly induce T cell anergy in vitro, therefore, application of imDC before and after skin transplantation probably can help relieve the rejection and prolong skin allografts survival. In the gene therapy based on imDC, it is necessary to transfer gene of interest such as CCR-7 or CTLA-4Ig into imDCs to achieve specific tolerance. However, some means of transfection and vectors often lead to imDC's maturation and change the important tolerance inducing functions of imDCs.In this study, imDCs were generated from human cord blood monocytes with rhGM-CSF and rhIL-4 for 7 days and imDCs were transfected with two different eucaryotic cell transfection system(Lipofectamine2000 and AdEasy) respectively on the 7th culture day. The cells in suspension were analysed in morphology, phenotype and immune functional tests. Furthermore, transfected cells were also applicated with IL-10 to evaluate... |
PDF Full Text Request