[Expression of nuclear factor-κB and its regulated cytokines in dendritic cells in patients with chronic severe hepatitis B and its clinical significance].
OBJECTIVE
To investigate the correlation of nuclear factor-κB (NF-κB) level and its regulated cytokines in monocyte-derived dendritic cells (MoDC) with illness severity and prognosis in patients with chronic severe hepatitis B (CSHB).


METHODS
Peripheral blood mononuclear cells (PBMC) were isolated by Ficoll density gradient separation from 37 patients with CSHB, 20 chronically HBV-infected patients, and 20 normal controls. Monocytes were acquired using immunomagnetic anti-CD14-beads. Next, monocytes were induced into immature DC (iDC) in vitro. On day six, polyI:C was added to induce DC maturation. Then mature DCs (mDCs) were collected at 48 h after polyI:C treatment to test the expression of NF-κB p50 by real time PCR. The supernatants were also collected respectively. The expression of TNFα and IL-6 in the supernatants were measured by ELISA.


RESULTS
The expression of NF-κB p50 in mDCs of patients with CSHB was the highest in three groups (F=19.01, P<0.01). The secretion of TNFα and IL-6 in mDCs from group CSHB was extremely high when compared with the other two groups, the secretions of TNFα in groups CSHB, CHB and NC were(15317.69+/-4124.90) pg/ml, (9670.29+/-3654.68) pg/ml and (6547.43+/-1027.20) pg/ml (F=45.77, P<0.01) respectively, and the productions of IL-6 in groups CSHB, CHB and NC were (1423.78+/-375.14) pg/ml, (862.68+/-93.68) pg/ml and (567.26+/-167.04) pg/ml (F = 67.60, P is less than 0.01), respectively. NF-κB p50 showed significant correlations with TNFα(r=0.52, P<0.01) and IL-6 (r=0.65, P<0.01) in mDCs. Furthermore, the secretions of TNFα and IL-6 in mDCs from group CSHB were negatively associated with PTA (r=-0.41, P=0.01; r=-0.40, P=0.01), but not with ALT, TBil and virus loads (r=-0.03, P=0.85, r=0.01, P=0.93, r=0.01, P=0.95; r=-0.09, P=0.58, r=0.16, P=0.34, r=0.09, P=0.59). No significant difference found in the expression of TNFα and IL-6 between the survival subgroup and the death subgroup in group CSHB (t=0.42, P=0.67; t=0.76, P=0.45).


CONCLUSIONS
The expression levels of NF-κB and its regulated cytokines in monocyte-derived DCs in patients with chronic severe hepatitis B were up-regulated and perhaps associated positively with the illness severity.
Qian
Zhi-ping
ZP
Department of Severe Hepatitis, Shanghai Public Health Clinic Center, Fudan University, and Department of Infectious Diseases, Huashan Hospital, Shanghai 201508, China.
Li
Ning
N
Zhu
Meng-qi
MQ
Zheng
Jian-ming
JM
Zhang
Yu-jie
YJ
Li
Qian
Q
Chen
Ming-quan
MQ
Wang
Jie-fei
JF
Shi
Guang-feng
GF
D000328:Adult; D016022:Case-Control Studies; D016207:Cytokines / Q000378:metabolism*; D003713:Dendritic Cells / Q000378:metabolism*; D005260:Female; D019694:Hepatitis B, Chronic / Q000097:blood / Q000378:metabolism*; D006801:Humans; D066298:In Vitro Techniques; D008297:Male; D008875:Middle Aged; D016328:NF-kappa B / Q000378:metabolism*
D016207:Cytokines; D016328:NF-kappa B