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Objective To identify the property and implications of dendritic cells of peripheral blood monocytes (PBMC) in patients with persistent HBV infection. Methods The PBMC from 18 patients with persistent HBV infection and 10 healthy subjects were incubated and induced into mature dendritic cells (DC) in the completed medium containing GM CSF、IL 4、FLt3 L and TNF ? cytokines. The flow cytometric analysis was employed to detect the expression of surface markers on DC. ELISA test was used to determine the level of interleukin 12 (IL 12) and IL 10 cytokines produced by DCs. The stimulatory capacity of DC were also determined in allogenic mixed leukocyte reaction (MLR). Results A typical morphology of DCs was observed when the PBMCs from healthy subjects and HBV infected patients were induced for 6 7 day in vitro incubation, but there were some significant differences of DCs as follows: (1) the proliferation ability and number of DCs significantly decreased, in particular, the expression levels of HLA DR,CD80(B7 1),CD86(B7 2) and CD1? on DC surface were simultaneously much lower in patients compared to healthy subjects ( P
RESUMO
Objective To investigate T cell-mediated anti-HBV effects of dendritic cells (DCs) sensitized by HBsAg in patients with HBV infection in vitro. Methods DCs isolated from peripheral blood mononuclear cells of HBV patients were cultured and proliferated in vitro, and then were stimulated with pure HBsAg before DCs maturation, and co-cultured with auto-T lymphocytes after DCs maturation. The T lymphocytes were then harvested 5 days later and added into the supernatant of 2.2.15 cell cultures. The supernatants were collected on 1st, 3rd, 5th and 7th day, respectively. ELISA was employed to detect HBeAg and the secretion of HBsAg, as well as to measure the concentration of IFN-? and IL-12 in culture supernatents. Results The proliferation and cytotoxicity of auto-T cells were markedly enhanced in the antigen stimulated DCs group compared with that of DCs without loaded antigen and only T lymphocyte group (P