AIM: To investigate the expression of nucleoporin 88 (Nup88) in hepatitis B virus (HBV) and C virus (HCV)-related liver diseases. METHODS: We generated a new monoclonal Nup88 antibody to investigate the Nup88 protein ...AIM: To investigate the expression of nucleoporin 88 (Nup88) in hepatitis B virus (HBV) and C virus (HCV)-related liver diseases. METHODS: We generated a new monoclonal Nup88 antibody to investigate the Nup88 protein expression by immunohistochemistry (IHC) in 294 paraffin-embedded liver specimens comprising all stages of hepatocellular carcinogenesis. In addition, in cell culture experiments HBV-positive (HepG2.2.15 and HB611) and HBV-negative (HepG2) hepatoma cell lines were tested for the Nup88 expression by Western-immunoblotting to test data obtained by IHC.RESULTS: Specific Nup88 expression was found in chronic HCV hepatitis and unspecific chronic hepatitis, whereas no or very weak Nup88 expression was detected in normal liver. The Nup88 expression was markedly reduced or missing in mild chronic HBV infection and inversely correlated with HBcAg expression. Irrespective of the HBV- or HCV-status, increasing Nup88 expression was observed in cirrhosis and dysplastic nodules, and Nup88 was highly expressed in hepatocellular carcinomas. The intensity of Nup88 expression significantly increased during carcinogenesis (P < 0.0001) and correlated with dedifferentiation (P < 0.0001). Interestingly, Nup88 protein expression was significantly downregulated in HBV-positive HepG2.2.15 (P < 0.002) and HB611 (P < 0.001) cell lines as compared to HBV-negative HepG2 cells. CONCLUSION: Based on our immunohistochemical data, HBV and HCV are unlikely to influence the expression of Nup88 in cirrhotic and neoplastic liver tissue, but point to an interaction of HBV with the nuclear pore in chronic hepatitis. The expression of Nup88 in nonneoplastic liver tissue might reflect enhanced metabolic activity of the liver tissue. Our data strongly indicate a dichotomous role for Nup88 in non-neoplastic and neoplastic conditions of the liver.展开更多
The human myxovirus resistance 2(Mx2/Mx B)protein,a member of interferon(IFN)-inducible dynamin-like large GTPases,restricts a number of virus infections.Inhibition of these viruses occurs at poorly-defined steps afte...The human myxovirus resistance 2(Mx2/Mx B)protein,a member of interferon(IFN)-inducible dynamin-like large GTPases,restricts a number of virus infections.Inhibition of these viruses occurs at poorly-defined steps after viral entry and has a common requirement for Mx B oligomerization.However,the GTPase activity is essential for the anti-viral effects of Mx B against herpesviruses and HBV but not HIV-1.To understand the role of Mx B GTPase activity,including GTP binding and GTP hydrolysis,in restriction of HIV-1 infection,we genetically separated these two functions and evaluated their contributions to restriction.We found that both the GTP binding and hydrolysis function of Mx B involved in the restriction of HIV-1 replication.The GTPase activity of Mx B contributed to its nuclear location,interaction with nucleoporins(NUPs)and HIV-1 capsids.Furthermore,Mx B disrupted the association between NUPs and HIV-1 cores dependently upon its GTPase activity.The function of GTPase activity was therefore multi-faceted,led to fundamentally distinct mechanisms employed by wild-type Mx B and GTPase activity defective Mx B mutations to restrict HIV-1 replication.展开更多
目的探讨核孔蛋白160(NUP160)基因对肾病综合征小鼠肾脏辅助性T细胞17(Th17)/调节性T细胞(Treg)免疫平衡的影响。方法将30只小鼠随机分为空白组(n=10)和造模组(n=20)。造模组小鼠经尾静脉注射阿霉素溶液(7.5 mg·kg-1)制备原发性肾...目的探讨核孔蛋白160(NUP160)基因对肾病综合征小鼠肾脏辅助性T细胞17(Th17)/调节性T细胞(Treg)免疫平衡的影响。方法将30只小鼠随机分为空白组(n=10)和造模组(n=20)。造模组小鼠经尾静脉注射阿霉素溶液(7.5 mg·kg-1)制备原发性肾病综合征(PNS)模型,空白组小鼠经尾静脉注射等容积的生理盐水。造模成功后将造模组小鼠随机分为PNS组和NUP160组,每组10只。NUP160组小鼠经尾静脉注射NUP160(10 mg·kg-1),每日1次;PNS组和空白组小鼠经尾静脉注射等量生理盐水,每日1次;3组小鼠均连续干预4周。给药0、2、4周时,应用生化自动检测仪检测小鼠24 h尿蛋白定量(24 h HUPQ);给药结束后,采用酶联免疫吸附试验法检测各组小鼠血清白细胞介素(IL)-17和IL-10水平;苏木精-伊红(HE)染色、过碘酸希夫(PAS)染色、Masson染色法观察各组小鼠肾组织病理学变化;流式细胞术检测各组小鼠肾组织中Th17、Treg细胞比例。结果给药0、2、4周时,PNS组和NUP160组小鼠24 h HUPQ显著高于空白组(P<0.05)。给药0周时,PNS组与NUP160组小鼠24 h HUPQ比较差异无统计学意义(P>0.05);给药2、4周时,NUP160组小鼠24 h HUPQ显著低于PNS组(P<0.05)。与空白组比较,PNS组和NUP160组小鼠血清IL-17水平显著升高,IL-10水平显著降低(P<0.05);与PNS组比较,NUP160组小鼠血清IL-17水平显著降低,IL-10水平显著升高(P<0.05)。空白组小鼠肾组织无明显病理学变化。与空白组相比,PNS组小鼠HE染色发现肾小球细胞排列紊乱,肾小球轻度萎缩,少许炎症细胞浸润,球囊壁轻度增厚,小球囊腔轻度扩张;PAS染色发现系膜出现增生,肾间质可见炎症细胞浸润;Masson染色发现小鼠肾小球基底膜和间质蓝色胶原沉积明显增多,组织纤维化程度显著升高。与PNS组相比,NUP160组小鼠肾组织以上症状均得到明显改善。与空白组比较,PNS组和NUP160组小鼠肾组织中Th17占比、Th17/Treg比值显著增加,Treg占比显著降低(P<0.05);与PNS组比较,NUP160组小鼠肾组织中Th17占比、Th17/Treg比值显著降低,Treg占比显著增加(P<0.05)。结论NUP160可调节PNS小鼠的Th17/Treg免疫平衡,进而发挥改善PNS的作用。展开更多
文摘AIM: To investigate the expression of nucleoporin 88 (Nup88) in hepatitis B virus (HBV) and C virus (HCV)-related liver diseases. METHODS: We generated a new monoclonal Nup88 antibody to investigate the Nup88 protein expression by immunohistochemistry (IHC) in 294 paraffin-embedded liver specimens comprising all stages of hepatocellular carcinogenesis. In addition, in cell culture experiments HBV-positive (HepG2.2.15 and HB611) and HBV-negative (HepG2) hepatoma cell lines were tested for the Nup88 expression by Western-immunoblotting to test data obtained by IHC.RESULTS: Specific Nup88 expression was found in chronic HCV hepatitis and unspecific chronic hepatitis, whereas no or very weak Nup88 expression was detected in normal liver. The Nup88 expression was markedly reduced or missing in mild chronic HBV infection and inversely correlated with HBcAg expression. Irrespective of the HBV- or HCV-status, increasing Nup88 expression was observed in cirrhosis and dysplastic nodules, and Nup88 was highly expressed in hepatocellular carcinomas. The intensity of Nup88 expression significantly increased during carcinogenesis (P < 0.0001) and correlated with dedifferentiation (P < 0.0001). Interestingly, Nup88 protein expression was significantly downregulated in HBV-positive HepG2.2.15 (P < 0.002) and HB611 (P < 0.001) cell lines as compared to HBV-negative HepG2 cells. CONCLUSION: Based on our immunohistochemical data, HBV and HCV are unlikely to influence the expression of Nup88 in cirrhotic and neoplastic liver tissue, but point to an interaction of HBV with the nuclear pore in chronic hepatitis. The expression of Nup88 in nonneoplastic liver tissue might reflect enhanced metabolic activity of the liver tissue. Our data strongly indicate a dichotomous role for Nup88 in non-neoplastic and neoplastic conditions of the liver.
基金supported by the National Science Foundation of China(81271818 and 81471940 to YF,and 81471941,81871659 and 81828005 to WH)
文摘The human myxovirus resistance 2(Mx2/Mx B)protein,a member of interferon(IFN)-inducible dynamin-like large GTPases,restricts a number of virus infections.Inhibition of these viruses occurs at poorly-defined steps after viral entry and has a common requirement for Mx B oligomerization.However,the GTPase activity is essential for the anti-viral effects of Mx B against herpesviruses and HBV but not HIV-1.To understand the role of Mx B GTPase activity,including GTP binding and GTP hydrolysis,in restriction of HIV-1 infection,we genetically separated these two functions and evaluated their contributions to restriction.We found that both the GTP binding and hydrolysis function of Mx B involved in the restriction of HIV-1 replication.The GTPase activity of Mx B contributed to its nuclear location,interaction with nucleoporins(NUPs)and HIV-1 capsids.Furthermore,Mx B disrupted the association between NUPs and HIV-1 cores dependently upon its GTPase activity.The function of GTPase activity was therefore multi-faceted,led to fundamentally distinct mechanisms employed by wild-type Mx B and GTPase activity defective Mx B mutations to restrict HIV-1 replication.
基金The Work Was Supported by the National Natural Science Foundation of China, the National Key Basic Research Project (No.2004CB117400),the Program for New Century Excellent Talents in University and the Key Project of Chinese Ministry of Education.
文摘目的探讨核孔蛋白160(NUP160)基因对肾病综合征小鼠肾脏辅助性T细胞17(Th17)/调节性T细胞(Treg)免疫平衡的影响。方法将30只小鼠随机分为空白组(n=10)和造模组(n=20)。造模组小鼠经尾静脉注射阿霉素溶液(7.5 mg·kg-1)制备原发性肾病综合征(PNS)模型,空白组小鼠经尾静脉注射等容积的生理盐水。造模成功后将造模组小鼠随机分为PNS组和NUP160组,每组10只。NUP160组小鼠经尾静脉注射NUP160(10 mg·kg-1),每日1次;PNS组和空白组小鼠经尾静脉注射等量生理盐水,每日1次;3组小鼠均连续干预4周。给药0、2、4周时,应用生化自动检测仪检测小鼠24 h尿蛋白定量(24 h HUPQ);给药结束后,采用酶联免疫吸附试验法检测各组小鼠血清白细胞介素(IL)-17和IL-10水平;苏木精-伊红(HE)染色、过碘酸希夫(PAS)染色、Masson染色法观察各组小鼠肾组织病理学变化;流式细胞术检测各组小鼠肾组织中Th17、Treg细胞比例。结果给药0、2、4周时,PNS组和NUP160组小鼠24 h HUPQ显著高于空白组(P<0.05)。给药0周时,PNS组与NUP160组小鼠24 h HUPQ比较差异无统计学意义(P>0.05);给药2、4周时,NUP160组小鼠24 h HUPQ显著低于PNS组(P<0.05)。与空白组比较,PNS组和NUP160组小鼠血清IL-17水平显著升高,IL-10水平显著降低(P<0.05);与PNS组比较,NUP160组小鼠血清IL-17水平显著降低,IL-10水平显著升高(P<0.05)。空白组小鼠肾组织无明显病理学变化。与空白组相比,PNS组小鼠HE染色发现肾小球细胞排列紊乱,肾小球轻度萎缩,少许炎症细胞浸润,球囊壁轻度增厚,小球囊腔轻度扩张;PAS染色发现系膜出现增生,肾间质可见炎症细胞浸润;Masson染色发现小鼠肾小球基底膜和间质蓝色胶原沉积明显增多,组织纤维化程度显著升高。与PNS组相比,NUP160组小鼠肾组织以上症状均得到明显改善。与空白组比较,PNS组和NUP160组小鼠肾组织中Th17占比、Th17/Treg比值显著增加,Treg占比显著降低(P<0.05);与PNS组比较,NUP160组小鼠肾组织中Th17占比、Th17/Treg比值显著降低,Treg占比显著增加(P<0.05)。结论NUP160可调节PNS小鼠的Th17/Treg免疫平衡,进而发挥改善PNS的作用。
