| Objective Our study attempts to investigate the characteristic of phenotype and function of hepatitis B virus antigens-pulsed myeloid dendritic cells derived from chronically HBV-infected patients with HCC. Methods PBMC from 31 CHB patients with concomitant HCC were collected by leukapheresis. mDC were propagated from PBMC in the serum-free AIM-V medium in the presence of cytokine cocktail (GM-CSF, IL-4, TNF-α, IL-1β , IL-6, PGE2) and pulsed with HBcAg or HBsAg, or LPS as control. After 8-d incubation, the phenotypic patterns of mDC were characterized by flow cytometry, and the levels of IL-10 and IL-12 produced by mDC were analyzed by ELISA. Autologous T-cell proliferation induced by mDC was tested by non-radioactive cell proliferation assay. IFN-γ from antigen-specific T cells was measured by ELISPOT assay. HBV-epitope-specific CD8+ T cells primed by mDC pulsed with the antigens were analyzed by Tetramer staining. Results 1. The surface markers of mDC: the expressions of CD80, CD83 on immature mDC were lower than that on mature mDC (CD80: 9.3%±3.3% vs 34.0%±20.7%, CD83: 4.5%±2.7% vs 13.2%±6.5%, both P<0.05), the expressions of CD80, CD86, CD40 and HLA-DR on HBcAg-pulsed mDC were significantly higher than that on unpulsed mDC (CD80: 58.3%±21.1% vs 34.0%±20.7%, CD86: 79.8%±12.9% vs 45.2%±22.0%, CD40: 69.8%±12.8% vs 54.2%±23.2%, HLA-DR: 78.4%±11.7% vs 54.6%±20.7%, all P<0.05), the expressions of CD86, HLA-DR on HBsAg-pulsed mDC were higher than that on unpulsed mDC (CD86:70.2%±9.1% vs 45.2%±22.0%, HLA-DR: 72.3%±3.9% vs 54.6%±20.7%, both P<0.05), the expressions of CD80, CD86, CD40, CD83, and... |
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