The mechanism of chronicity of hepatitis B (inhibition of antigen presentation by HBX protein)

乙型肝炎慢性化机制（HBX蛋白抑制抗原呈递）

• 批准号: 17590631
• 负责人: MURATA Kazumoto
• 金额: $ 2.24万
• 依托单位: Jichi Medical University (2006)Mie University (2005)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-17590631/
• 关键词: Proteasome; HBX protein; Vaccinia virus; Adeno virus; CTL assay; proteasome

First, the expression of HBX protein was confirmed in adeno-HBX infected Renca cells (Balb/c mouse syngenic cell line), but not in adeno-HBX0 (HBX knockout) infected cells. Then, we examined MHC class I expression on the Renca cells infected with adeno-HBX or adeno-HBX0 (M.O.I.=0.5) 2 days after infection by flow cytometer. Its expression on the adeno-HBX infected Renca cells were significantly inhibited in comparison with that on the adeno-HBX0 infected Renca cells. Next, we tried to check the effect of HBX on cytotoxic lymphocyte (CTL) responses in vivo. We generated adeno-specific CTL clone by injection of adeno-LacZ to Balb/c mice intraperitoneally every 2 weeks, following ex vivo culture with fresh adeno-LacZ infected-dendritic cells for 7 days. A standard CTL assay was performed, using adeno-HBX infected Renca cells or adeno-HBX0 infected Renca cells as target cells. Chromium releasing was inhibited when we used adeno-HBX infected Renca cells as target cells, compared with adeno-HBX0 infected Renca cells. These results were quantitatively confirmed by intracellular cytokine (IFN-γ) staining. WE also examined these, using Vaccinia virus (vvHBX or vvHBX0). Both in vitro and in vivo study showed the same tendencies as using adeno-viruses. These pieces of evidences suggest that HBX inhibits antigen presentation through inhibition of proteasome. From these results, we speculate that HBX inhibits antoigen presentation through inhibition of proteasome, following immune escape.

首先，在腺病毒-HBX感染的Renca细胞（Balb/c小鼠同基因细胞系）中证实了HBX蛋白的表达，但在腺病毒-HBX 0（HBX敲除）感染的细胞中没有。然后，我们检测了用腺病毒-HBX或腺病毒-HBX 0感染的Renca细胞上MHC I类分子的表达（M.O.I.= 0.5)感染后2天用流式细胞仪检测。Adeno-HBX感染的Renca细胞中其表达明显低于Adeno-HBX 0感染的Renca细胞。接下来，我们试图检查HBX对体内细胞毒性淋巴细胞（CTL）反应的影响。我们通过每2周向Balb/c小鼠腹腔注射腺病毒LacZ，然后与新鲜的腺病毒LacZ感染的树突状细胞体外培养7天来产生腺病毒特异性CTL克隆。使用腺病毒-HBX感染的Renca细胞或腺病毒-HBX 0感染的Renca细胞作为靶细胞进行标准CTL测定。以Adeno-HBX感染的Renca细胞为靶细胞，与Adeno-HBX 0感染的Renca细胞相比，铬的释放受到抑制。这些结果通过细胞内细胞因子（IFN-γ）染色定量证实。我们还使用牛痘病毒（vvHBX或vvHBX 0）对这些进行了检查。体外和体内研究均显示出与使用腺病毒相同的趋势。这些证据表明，HBX通过抑制蛋白酶体抑制抗原呈递。根据这些结果，我们推测HBX通过抑制蛋白酶体抑制抗原呈递，随后免疫逃逸。

项目成果

Cholangiocarcinoma with chylous ascites.

胆管癌伴乳糜性腹水。

• 发表时间: 2005
• 期刊: J Gastroenterol Hepatol 19
• 作者: Osawa S;Ikuma M et al.;Yamamoto N
• 通讯作者: Yamamoto N

Cardiac metastases of gallbladder carcinoma.

胆囊癌心脏转移。

• 发表时间: 2005
• 期刊: World J Gastroenterol 11
• 作者: Abe H;et al.;Inoue T
• 通讯作者: Inoue T

Up-regulation of IL-18 by interferon alpha/ribavirin combination therapy induces an anti-viral effect in patients with chronic hepatitis C.

干扰素α/利巴韦林联合疗法上调 IL-18 可在慢性丙型肝炎患者中产生抗病毒作用。

• 发表时间: 2005
• 期刊: Hepato=Gastroenterol 11
• 作者: Keiichi Kiriya;et. al.;Nanakin A;Tsutsui S;Sugimoto M;Murata K
• 通讯作者: Murata K

Up-regulation of IL-18 by IFN alpha/ribavirin combination therapy induces an anti-viral effect in patients with chronic hepatitis C

IFNα/利巴韦林联合疗法上调 IL-18 可在慢性丙型肝炎患者中产生抗病毒作用

• 发表时间: 2005
• 期刊: Hepato-Gastroenterology 52
• 作者: Murata K;Yamamoto N;Kawakita T;et al.
• 通讯作者: et al.

Tumor thrombi in esophageal varices.

食管静脉曲张出现癌栓。

• 发表时间: 2005
• 期刊: J Gastroenterol Hepatol 20
• 作者: Nanakin A;Fukui H;Fujii S;Sekikawa A;Kanda N;Hisatsune H;Seno H;Konda Y;Fujimori T;Chiba T.;Sugimoto K
• 通讯作者: Sugimoto K