[Effects of CpG-ODN combined with HBsAg on the phenotype, function and the activity of NF-kappa B and AP-1 of monocyte-derived dendritic cells in chronic hepatitis B patients].
OBJECTIVE
To investigate the effect of synthetic non-methylated CpG-ODN combined with recombined HBsAg on the phenotype, function and the expression level of nucleic transcription factor NF-kappa B (NF-kB) and AP-1 of monocyte-derived dendritic cells (DC) in patients with chronic hepatitis B (CHB).


METHODS
Purified monocytes were isolated from the peripheral blood of CHB patients and healthy volunteers and cultured with human granulocyte-monocyte colony stimulating factor added together with interleukin-4. On the fifth day of culture, CpG-ODN, HBsAg and other reagents, such as TNF alpha and PBS, were added to the medium, and 18 hours later cells were collected for the detection of surface molecules (HLA-DR/CD86/CD1a). IL-12p70 levels in the supernatant and stimulating capacity to allogenic T lymphocytes were detected. The nucleic proteins of NF-kB and AP-1 in DC were extracted and purified for the gel shift assay.


RESULTS
Compared with those of the PBS group, the expression rates of HLA-DR of DC treated with CpG-ODN and/or HBsAg were obviously increased. Both the IL-12p70 level and the stimulating capacity of DC to allogenic T lymphocytes in mixed lymphocyte reaction increased significantly in the CpG-ODN group and in the CpG-ODN/HBsAg combination group (P less than 0.01 and 0.05, respectively). The expression rates of CD1a were raised only in the CpG-ODN/HBsAg combination group. Three kinds of immunological adjuvants, TNF alpha, CpG-ODN and CpG-ODN/HBsAg enhanced the expression of nucleic NF-kB and inhibited the expression of AP-1 in DC.


CONCLUSIONS
CpG-ODN, like TNF alpha, has remarkable stimulatory effect on the impaired phenotype and function of monocyte-derived DC in patients with CHB; CpG-ODN and HBsAg have a synergetic effect in increasing the antigen presenting function. The regulating ability of CpG-ODN and TNF alpha on the expression levels of NF-kB and AP-1 might be one of the mechanisms of their immunostimulatory effects on DC of the CHB patients.
Xiang
Xiao-Xing
XX
Department of Digestive Diseases, Subei People's Hospital of Jiangsu Province, Yangzhou 225001, China. xiangxx1965@yahoo.com.cn
Zhou
Xia-Qiu
XQ
Xie
Qing
Q
Yu
Hong
H
Zhou
Hui-Juan
HJ
D000276:Adjuvants, Immunologic / Q000494:pharmacology; D000328:Adult; D002478:Cells, Cultured; D003713:Dendritic Cells / Q000378:metabolism*; D006684:HLA-DR Antigens / Q000378:metabolism; D006514:Hepatitis B Surface Antigens / Q000494:pharmacology*; D019694:Hepatitis B, Chronic / Q000378:metabolism*; D006801:Humans; D008297:Male; D016328:NF-kappa B / Q000378:metabolism; D009838:Oligodeoxyribonucleotides / Q000494:pharmacology*; D011993:Recombinant Fusion Proteins; D018808:Transcription Factor AP-1 / Q000378:metabolism*
D000276:Adjuvants, Immunologic; C408982:CPG-oligonucleotide; D006684:HLA-DR Antigens; D006514:Hepatitis B Surface Antigens; D016328:NF-kappa B; D009838:Oligodeoxyribonucleotides; D011993:Recombinant Fusion Proteins; D018808:Transcription Factor AP-1