The Study Of The Dendritic Cell Function In The Patients With Chronic Hepatitis B Failed Response To Lamivudine

Objectives: To compare the function of dendritic cell among the patients with chronic hepatitis B response or no response to lamivudine , and healthy persons . To study whether the function of dendritic cell could be improved with the present of type I interferon or hepatitis B surface antigen .Methods: The 10mL peripheral blood of every enrolled people including 4 healthy persons, 5 responders and 7 non-responders to lamivudine was collected and separated peripheral blood mononuclear cells (PBMC) which would be divide into T lymphocytes and monocytes. The separated T lymphocytes were stored in liquid nitrogen system and IL-4 and GM-CSF were used to stimulate and transform these monocytes to be immature dendritic cell (imDC) in 7 days culture. Then some imDCs based on the stimulating of TNFa were cultured in 24 holes cell culture plate mixed with HLA compatible T lymphocytes and each of 4 different circumstances by using Hepatitis B Virus surface antigen and interferon a-1b. And the other imDCs based on the stimulating of TNFa cultured in 75cm~2 cell culture bottle. 5 days later, the co-stimulate molecule CD80 and CD86 of cells cultured in 24 holes cell culture plate were analyzed by flow cytometry technology and cytokines including IL12p70, 1L10 and IFNγ detected by ELISA method. HLADR, CD80 and CD86 of cells cultured in 75cm~2 cell culture bottle were analyzed by flow cytometry technology compared with PBMC to prove mature DC gathered. Then these mature DC mixed HLA compatible Tlymphocytes cultured 5 days and the stimulating index(SI) of mixed lymphocyte reaction(MLR) was detected with MTT method.Results: IL - 12p70, the main cytokine secreted by dendritic cells, will be elevated by the stimulating of both Hepatitis B Virus surface antigen and interferon a - lb in all enrolled people. But the degree of this elevation in healthy individuals is higher than those chronic hepatitis B patients whether they are responders to lamivudine or not. Hepatitis B Virus surface antigen alone can reduce the production of IL - 12p70 in healthy persons while not in chronic hepatitis B patients. The amount of the production of IL - 12p70 are indifferent between healthy persons and responders to lamivudine during the stimulating of TNFa only . But it is lower in non-responders in the same circumstance. The production of IFNy is highest in healthy individuals and lowest in non-responders among the three groups in the each of four circumstances. And it is also elevated by the stimulating of both Hepatitis B Virus surface antigen and interferon a - 1 b in the each of three groups . The amount of the production of IL - 10 will be elevated by the stimulating of both Hepatitis B Virus surface antigen and interferon a - lb only in non-responders but not in healthy people and responders. There is no difference among the three groups of the production of IL - 10. In any circumstances, the SI of MLR in healthy people is higher than responder and non-responders'. When the ratio of DC: T is higher ,0.1 or 0.05 in this study, the SI of MLR of responders is higher than non-responders'. But when the ratio of DC: T is lower, 0.025 or 0.0125 in this study , there is no difference of the SI of MLR between non-responders and responders. The expression of co-stimulate molecule CD86 could be induced by the stimulating of both Hepatitis B Virus surface antigen and interferon a - lb in healthy people and responders, but not in non-responders. The expression of CD80 and CD86 in healthy people is higher than other two groups during the stimulating of TNFa only. While both Hepatitis...