Objective: To compare the phenotype and function of dendritic cells (DCs) of patients with chronic hepatitis Beta with those of DCs from healthy persons.
Method: Peripheral blood monocytes (PBMCs) were isolated from 10 patients with chronic hepatitis Beta and 10 healthy blood donors. DCs were generated by culturing PBMCs in 1 000 U/ml granulocyte-macrophage stimulating factor (GM-CSF) and 500 U/ml IL-4 with AIM-V in vitro. DCs and supernatant were collected 3, 5, 7, 9, 11, 13 and 15 days after culture. TNF- alpha 50 ng/ml was added into the culture of DCs from patients with hepatitis Beta and DCs were collected 48 hours after. FACS was used to detect the phenotype. The changes of cell phenotype on different days with or without TNF-alpha were tested by immunolabelling and flow cytometry analysis. IL-12 ELISA kit was applied to investigate IL-12 secretion of DCs. To perform mixed leucocyte reaction (MLR) test, DCs were cocultured with allogeneic T cells at different stimulatory ratio.
Results: The expression of CD83 was higher in the group with TNF-alpha than in the group without TNF-alpha. The positive rates of CD80 and CD54 were not different between the two groups. DCs from patients and normal donors with typical morphology were harvested successfully in vitro. The IL-12 level was 103.93 +/- 12.59 pg/ml in the group with TNF-alpha, higher than that in the group without TNF-alpha (11.45 +/- 2.58 pg/ml, P < 0.05). MLR test showed that when the stimulatory ratio of allogeneic T cells was 1 : 20, 1 : 50, and 1 : 100 respectively, the results were 40 835 +/- 3 480, 37 525 +/- 6 398, and 31 814 +/- 5 521 respectively in group of patient; and 49 097 +/- 2 273, 43 049 +/- 2 833, and 39 091 +/- 6 469 respectively in the group of healthy blood donors.
Conclusion: 1.DCs of patients and of healthy persons can be cultured success fully with AIM-V when induced by GM-CSF, IL-4 and TNF-alpha in vitro. 2. Functions of DCs from patients are lower than those from healthy donors. However, they are not different in amount and morphology.