[Phenotype and function of myeloid dendritic cells pulsed with hepatitis B virus antigens in patients with HBV-associated hepatocellular carcinoma].
OBJECTIVE
To investigate the characteristics of phenotype and function of myeloid dendritic cells (mDCs) pulsed with HBV antigens derived from HBV-associated hepatocellular carcinoma (HCC) patients.


METHODS
Peripheral blood mononuclear cells (PBMCs) were collected, using blood cell separator, from 21 primary HCC patients and 4 healthy donors. mDCs were propagated in serum-free AIM-V medium in the presence of cytokine cocktail, and pulsed with HBcAg or HBsAg. After 9 days' incubation, the phenotypic patterns of mDC were characterized by flow cytometry and the levels of IL-10 and IL-12 produced by mDCs were analyzed by ELISA. Autologous T cells proliferation stimulated by mDC was tested by non-radioactive cell proliferation assay kit.


RESULTS
The expression rates of CD80, CD86, CD40, and HLA-DR in the mDCs pulsed with HBcAg were 55% +/- 26%, 80% +/- 13%, 70% +/- 13% and 73% +/- 24% respectively, significantly higher than those of the control group (29% +/- 25%, 35% +/- 18%, 44% +/- 26% and 45% +/- 23% respectively, all P < 0.05). Among the expression rates of surface molecules of the mDCs pulsed with HBsAg only the expression rate of HLA-DR was significantly higher than that of the un-pulsed mDCs (63% +/- 15% vs 45% +/- 23%, P < 0.05). T cell proliferation assay revealed an impaired allostimulatory capacity of mDCs in HCC and the stimulatory capacity of the mDCs pulsed with HBcAg to induce proliferation of autologous T cells was more powerful than that pulsed with HBsAg (P < 0.05). The levels of IL-10 and IL-12 produced by the mDCs pulsed with HBsAg were (35 pg/ml +/- 9 pg/ml and 135 pg/ml +/- 63 pg/ml respectively, both significantly lower than those pulsed with HBcAg (236 pg/ml +/- 95 pg/ml and 733 pg/ml +/- 212 pg/ml respectively, both P < 0.05).


CONCLUSIONS
Pulsation of mDCs in vitro by HBcAg or HBsAg enhance the expression of CD80, CD86, CD40, and HLA-DR, and increase the ability of mDCs to stimulate the proliferation of autologous T lymphocytes.
Qian
Shen
S
Research Center of Biological Therapy, Hospital No. 302 of Chinese People's Liberation Army, Beijing 100039, China.
Shi
Ming
M
Zhang
Hui
H
Zhang
Bing
B
Xu
Dong-ping
DP
Wang
Fu-sheng
FS
D000328:Adult; D000368:Aged; D006528:Carcinoma, Hepatocellular / Q000150:complications / Q000276:immunology*; D003713:Dendritic Cells / Q000276:immunology* / Q000821:virology; D005260:Female; D006511:Hepatitis B Antigens / Q000276:immunology*; D019694:Hepatitis B, Chronic / Q000150:complications / Q000276:immunology*; D006801:Humans; D016130:Immunophenotyping; D008113:Liver Neoplasms / Q000150:complications / Q000276:immunology*; D008297:Male; D008875:Middle Aged
D006511:Hepatitis B Antigens