Qian Zhi-ping, Li Ning, Zhu Meng-qi, Zheng Jian-ming, Zhang Yu-jie, Li Qian, Chen Ming-quan, Wang Jie-fei, Shi Guang-feng
Department of Severe Hepatitis, Shanghai Public Health Clinic Center, Fudan University, and Department of Infectious Diseases, Huashan Hospital, Shanghai 201508, China.
Zhonghua Gan Zang Bing Za Zhi. 2010 Aug;18(8):590-4. doi: 10.3760/cma.j.issn.1007-3418.2010.08.010.
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).
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.
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).
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.
探讨慢性重型乙型肝炎（CSHB）患者单核细胞来源的树突状细胞（MoDC）中核因子-κB（NF-κB）水平及其调控的细胞因子与病情严重程度及预后的相关性。
采用Ficoll密度梯度离心法从37例CSHB患者、20例慢性HBV感染者及20例正常对照者中分离外周血单个核细胞（PBMC）。利用免疫磁珠抗CD14法获取单核细胞。随后，将单核细胞在体外诱导为未成熟树突状细胞（iDC）。第6天，加入聚肌胞苷酸（polyI:C）诱导DC成熟。然后在polyI:C处理48小时后收集成熟树突状细胞（mDC），通过实时PCR检测NF-κB p50的表达。同时分别收集上清液。采用ELISA法检测上清液中TNFα和IL-6的表达。
CSHB患者mDC中NF-κB p50的表达在三组中最高（F=19.01，P<0.01）。与其他两组相比，CSHB组mDC中TNFα和IL-6的分泌极高，CSHB组、慢性乙型肝炎（CHB）组和正常对照（NC）组的TNFα分泌量分别为（15317.69±4124.90）pg/ml、（9670.29±3654.68）pg/ml和（6547.43±1027.20）pg/ml（F=45.77，P<0.01），IL-6的分泌量分别为（1423.78±375.14）pg/ml、（862.68±93.68）pg/ml和（567.26±167.04）pg/ml（F = 67.60，P<0.01）。mDC中NF-κB p50与TNFα（r=0.52，P<0.01）和IL-6（r=0.65，P<0.01）呈显著正相关。此外，CSHB组mDC中TNFα和IL-6的分泌与凝血酶原活动度（PTA）呈负相关（r=-0.41，P=0.01；r=-0.40，P=0.01），但与谷丙转氨酶（ALT）、总胆红素（TBil）及病毒载量无关（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）。CSHB组生存亚组与死亡亚组之间TNFα和IL-6的表达无显著差异（t=0.42，P=0.67；t=0.76，P=0.45）。
慢性重型乙型肝炎患者单核细胞来源的DC中NF-κB及其调控的细胞因子表达上调，可能与病情严重程度呈正相关。
