On The Positive Function Of The Dendritic Cells From Patients With Chronic Hepatitis B Plused With HBsAg To HBsAg-Specific Cytotoxicity T Lymphocytes

Objective: To study the function including immunophenotypic, immunologic function and anti-HBsAg specific cell-mediated immunity of monocytes-derived dendritic cell(DCs) plused with HBsAg from patients with chronic hepatitis B. To explore a new method that the DCs plused with HBsAg are used as immunologic adjuvant to get an efficient and HBsAg-special CTL response. Methods: DCs generating from periopheral blood monocytes are cultured in media containing rhGM-CSF(800u/ml), rhIL-4(500u/ml)and rhTNF- a (250u/ml). Then the DCs divided into two groups, one was the experimental group plused with HBsAg(10ng/ml)on the 4th day ; the other was the control group without HBsAg. On the 7th day, immunophenotype of DCs in the two groups including CD1a, CD80, CD83, CD86, CD40, HLA-DR was characterized by FACS and immunologic function of them was detected through the stimulating reaction of allogenic T lymphocytes. The concentration of cytokines in supernatant such as IL-6 and IL-12 was tested by ELISA .In the experiment, HBsAg were added into the DCs on the 4th day,then the DCs were co-cultured with T cells on the 5th day .On the 7th day, HBsAg was added into media. On the 10th day ,the CTL's response to HepG2 2.2.15cells,HepG2 cells and K562 cells were detected by LDH. Results: (l)Expression of the immunophenotype of the experimental groups including CD1a, CD80, CD83, CD86, CD40, HLA-DR was higer than that of the control group (p<0.01) (2)The concentration of IL-12 in the supernatant of the experimental groups washiger and IL-6 was much lower than that of the control groups (p<0.01)(3)The capacity of DCs stimulating allogenic lymphocyte to proliferate greatly was increased compared with that of control(p<0.01) (4)DCs plused with HBsAg could efficiently present HBsAg and induce T lymphocytes to differentiate into CTLs, which could efficiently kill HepG2 2.2.15cells that express HBsAg on the surface .The killing rate of the experimental groups (66.0% +2.7%) was much higer than that of the two control groups (p<0.01).But CTLs of the three groups have quite lower ability to kill hepatogenic HepG2 cells and non- hepatogenic K562 cells. Conclusion: DCs plused with HBsAg, which developed with rhGM-CSF, rhIL-4 and rhTNF- a , could greatly increase expression of immunophenotype of DCs (CD1a, CD80, CD83, CD86, CD40, HLA-DR). Meanwhile, DCs still enchance the capacity of stimulating allogenic lymphocyte to proliferate. The level of IL-12 secretion increased,but the level of IL-6 secretion decreased. Specical and efficient CTL response can be significantly induced by DCs plused with HBsAg. It will probably provide a experiemental evidence as immunotherapy for patients with Chronic hepatitis B.