/ 410007 HBV DCs DCs HBV PBMCs DCs DCs HLA-DR CD80 CD86 CD83 CD1 琢 DCs IFN- 琢 IL-4 HBV HBV DCs HLA-DR CD1 琢 CD83 CD80 CD86 P<0.01 DCs IFN- 琢 P<0.01 HBV CD83 CD86 P<0.01 IL-4 P<0.01 DCs IFN- 琢 P<0.01 HBV DCs HBV doi: 10.11842/wst.2013.05.003 R259 A 1 1.1 1.1.1 Dendritic cells DCs DCs 2012 1 ~2012 12 Acute-on-Chronic Liver Fail 原 ure ACLF Chronic LiverFailure CLF 2013-05-20 2013-06-04 ACLF 24 8 元 JDZX2012061 - - 81102594/H2708 DC T 2010FJ3018 HBV 2010022 元元 813
绎 4 32.5 依 8.2 12 6 6 35.7 依 6.9 B Prothrombin time activity percent 原 age PTA% P>0.05 8 5 3 27~45 33.5 依 5.8 1.1.2 2012 ACLF CLF 3~4 1.1.3 HCV HDV HAV HEV HGV EB 3 3 1.2 DCs DCs DCs HLA -DR CD1 琢 CD80 CD86 CD83 DCs IL-4 IFN- 琢 1.2.1 PBS 10% Fetalcalfserum FCS Hyclone RPMI-1640 CD1a -FITC HLA -DR -P CD80 -FITC CD86 -PE CD83 -FITC IgG2b IgG1-FITC ebioscience 4 rhil-4 Peprotech rhgm-csf Pe 原 protech 1.2.2 BD CO 2 XD- 202 1.2.3 Peripheral blood monouclear cells PBMCs 20 ml 50 U ml -1 PBS 10 ml 5mL 15 ml 45 毅 1cm 2 000 rpm 30 min 3 PBMCs PBMCs 5 PBS 2, 1 000 rpm 10 min 1mLPBS 1 1 0.2 豫 4 10% FCS RPMI-1640 PBMCs 2 伊 10 6 /ml 6 2.5 ml 37 益 5% CO 2 37 益 RPMI1640 2 6 2.5 ml 10% FCS RPMI -1640 814
rhgm -CSF 1 000 U ml -1 rhil -4 500 U ml -1 5%CO 2 37 益 1/2 7 DCs DCs 7 DCs 1500rpm 10 min PBS 2 1 000 rpm 10 min 2 2 伊 10 5 / 6 l.5 ml Ep 原 pendorf, 100 滋 L CD1a-FITC CD86-PE CD80-FITC CD83-FITC HLA-DR-FITC 20 滋 L 4 益 45 min PBS 2 1% 500 滋 L PE IgG CD1a HLA -DR CD80 CD86 CD83 DCs - 琢 IFN- 琢 -4 IL- 4 DCs 7 l.5 ml Eppendorf ABC-ELISA 1.3 SPSS 16.0 依 x 依 s t P<0.05 2 2.1 DCs DCs HLA -DR CD1 琢 CD83 CD80 CD86 P<0.01 1 2.2 DCs DCs IFN- 琢 P<0.01 IL-4 2 2.3 DCs DCs HLA -DR CD1 琢 CD80 CD83 CD86 P<0.01 P<0.05 HLA -DR CD1 琢 CD80 CD83 CD86 P<0.01 3 2.4 DCs DCs IFN - 琢 P<0.01 1 DCs HLA-DR CD1a CD1a CD80 CD86 24 80.70 依 4.04 22.78 依 4.38 23.56 依 3.72 44.13 依 7.16 67.37 依 1.02 8 90.45 依 4.37 44.57 依 3.12 40.29 依 4.17 78.92 依 2.18 85.02 依 3.93 P<0.01 2 DCs IFN- 琢 /pg ml -1 IL-4/pg ml -1 24 77.98 依 7.44 38.94 依 1.16 8 28.56 依 8.31 37.77 依 10.28 P<0.01 815
绎 IFN- 琢 P<0.01 IL-4 P<0.01 4 3 Liver failure 60%~80% ACLF CLF [1] 90% [2] [3 4] DCs T [5] [6] DCs-T HBV [7] CTL NK DCs DCs HBV [8] [9] / HBV DCs / HBV [10] HBV DCs 淤 HBV DCs HBV DCs 于 DCs 盂 DCs IFN- 琢 DCs IL-4 HBV HBV DCs CD83 CD86 / IFN- 琢 /IL-4 HBV 3 DCs HLA-DR CD1 琢 CD83 CD80 CD86 82.34 依 7.15 23.6 依 5.52 25.8 依 2.17 46.27 依 6.1 吟 74.03 依 10.57 80.12 依 2.03 22.39 依 2.08 绎 20.27 依 3.19 43.56 依 8.70 绎 61.96 依 12.03 8 90.45 依 4.37 44.57 依 3.12 40.29 依 4.17 78.92 依 2.18 85.02 依 3.93 吟绎 P<0.05 P<0.01 P<0.01 4 DCs 原 1 IFN- 琢 /pg ml 原 1 IL-4/pg ml 绎 98.07 依 13.21 绎 31.64 依 4.88 57.16 依 11.7 51.62 依 6.94 8 28.56 依 8.31 37.77 依 10.28 绎 P<0.01 P<0.01 816
DCs IFN- 琢 IL-4 1,. 2012.,2012,5(6) 颐 321~327. 2,,,. 1977.,2008,16(10) 颐 772~775. 3,..,2008,13(1) 颐 60~61. 4,,..,2007,12(S1) 颐 6~10. 5,.., 2009,14(1) 颐 53~56. 6,,,..,2008,16(9) 颐 660~664. 7 Royer P J, Bougras G, Ebstein F, et al. Efficient monocyte-derived dendritic cell generation in patients with acute myeloid leukemia af 原 ter chemotherapy treatment: application to active immunotherapy. Exp Hematol, 2008, 36(3) 颐 329~339. 8,..,2012,28(3) 颐 169~171. 9,,,..,2011,43(1) 颐 4~6. 10,,,..,2010,20(5) 颐 260 ~262. Comparison on Dendritic Cells Function of Yang-Huang and Yin-Huang Syndrome in HBV Related Hepatic Failure Patients Zhang Tao, Ji Jing, Ji Enru, Chen Bin, Huang Yuhong (The First A ffiliated Hospital of Hunan University of Chinese Medicine / National Clinical Research Base of Traditional Chinese Medicine for Liver Disease, Changsha 410007, China) Abstract: This study was aimed to compare the function of dendritic cells (DCs) in patients of Yin-Huang syn 鄄 drome and Y ang-huang syndrome from the peripheral blood of HBV related hepatic failure. Jaundice patients of HBV related hepatic failure were divided into two groups, which were the Yang-Huang syndrome group and Yin- Huang syndrome group. And a healthy control group was also established. The DCs were separated and cultured in vitro from PBMCs in the peripheral blood. Surface molecules such as HLA -DR, CD80, CD86, CD83 and CD1 琢 were detected by the flow cytometry. And the level of IFN- 琢 and IL-4 in the supernatant of DCs were also de 鄄 tected. The expression difference of inflammatory cytokines and immune cells between Yin-Huang and Yang- Huang syndrome in patients of HBV related hepatic failure were compared. The results showed that compared with healthy people, the expression rate of DCs phenotype such as HLA-DR, CD1 琢, CD83, CD80, CD86 in patients of HBV related hepatic failure was significantly decreased (P < 0.01); the excreted factor IFN- 琢 had a significant rising (P <0.01).ComparedtoYang-Huang syndrome group, the expression rate of CD83 and CD86 in the Yin- Huang syndrome group of HBV related hepatic failure patients was significantly reduced (P <0.01).Andtheex 鄄 creted factor of IL-4 had a significant rising in Yin-Huang syndrome group (P < 0.01). Compared to the Yin- Huang syndrome group, the excreted factor IFN- 琢 in the Yang-Huang syndrome group had a significant rising (P < 0.01). It was concluded that DCs function of patients with HBV related hepatic failure in both Yin-Huang syn 鄄 drome and Yang -Huang syndrome group showed low immune function. And the immune function of the Yin- Huang syndrome group was lower. There was excessive releasing of inflammatory factors in the Yang-Huang syn 鄄 drome group. And there was enhanced anti-inflammatory cytokine expression in the Yin-Huang group. Keywords: HBV-related liver failure, Yin-Huang syndrome, Y ang-huang syndrome, dendritic cells 817