戊型肝炎病毒抗原研究进展
作者：杜珩 庄辉
单位：100083 北京医科大学微生物系
关键词：
肝脏000132 戊型肝炎病毒(HEV)为线状单股正链RNA病毒，基因组全长约7.5kb(7.2～7.6kb)，编码2 400～2 533个氨基酸，由5’端非结构区(NS)和3’端结构区(S)组成，3’端带一个由150～200个腺苷酸残基组成的poly A尾巴。HEV基因组含3个开放读码框架(ORFs)，ORF1位于5’NS区，长5 079bp(第28～5107nt)，编码HEV复制相关蛋白，包括甲基转移酶、木瓜酶样的蛋白酶、解旋酶、RNA依赖性RNA聚合酶(RDRP)；ORF2从ORF1终止密码下游37个bp开始，长1 980bp(第5 147～ 7127nt)，编码660个氨基酸，主要包括信号肽和衣壳蛋白；ORF3位于ORF1、ORF2之间(第5 106～5 478nt)，与两者均有部分重叠，长369bp，编码一段123个氨基酸的多肽，功能不清。有学者认为ORF3编码的蛋白是一种与细胞骨架有关的磷蛋白，也有认为与转膜元件有关。本文仅就HEV抗原表位及ORF2抗原真核表达的研究进展简要综述如下。
, 百拇医药
一、HEV抗原表位的研究
(一)ORF1编码蛋白的抗原表位 Kaur等[1]在根据ORF1序列设计合成的系列多肽片段中，发现人类线状B细胞抗原表位存在于12个独立的多肽片段，其中有5个集中于第1207～1 446aa，该区域正处于RDRP内。RDRP内的抗原表位可能与非保护性反应有关；这些抗原也可能派生出附着于感染细胞膜表面的抗原位点，从而参与抗体依赖的抗病毒机制，如抗体依赖性细胞毒作用。……
参考文献：
[1]Kaur M,Hyams KC,Purdy MA,et al.Human linear B-cell epitopes encoded by the hepatitis E virus include determinants in the RNA-de-pendent RNA polymerase.Proc Natl Acad Sci USA,1992,89:3855-3858.
, 百拇医药
[2]Li F,Zhuang H,Kolivas S,et al.Persistent and transient antibody responses to hepatitis E virus detected by western immunoblot using open reading frame 2 and 3 and glutathione S-transferase fusion pro-teins.J Clin Microbiol,1994,32:2060-2066.
[3]Li F,Torresi J,Locarnini SA,et al.Amino-terminal epitopes are ex-posed when full-length open reading frame 2 of hepatits E virus is ex-pressed in Escherichia coil,but carboxyterrninal epitopes are masked.J Med Virol,1997,52:289-300.
, 百拇医药
[4]Purdy MA,McCaustland KA,Krawczynsk K,et al.Expression of a hepatitis E virus (HEV)-trpE fusion protein containing epitopes recognized by antibodies in sera from human cases and experimentally infected primates.Arch Virol,1992,123:335-349.
[5]Tsarev SA,Tsareva TS,Emerson SU,et al.ELISA for antibody to hepatitis E virus (HEV) based on complete open-reading frame-2 pro-tein expressed in insect cells:identification of HEV infection pri-mates.J Infect Dis,1993,168:369-378.
, 百拇医药
[6]Robinson RA,Burgess WH,Emerson SU,et al.Structural charac-terization of recombinant hepatitis E virus ORF2 proteins in bac-ulovirus-infected insect cells.Protein Exp Puri,1998,12:75-84.
[7]McAtee CP,Zhang YF,Yarbough PO,et al.Purification and characterization of a recombinant hepatits E protein vaccine candidate by liquid chromatography-mass spectrometry.J Chromatogr B Biomed Appl,1996,685:91-104.
[8]Li TC,Yamakawa Y,Suzuki K,et al.Expression and self-assembly of empty virus-like particles of hepatitis E virus.J Virol,1997,71:7207-7213.
, http://www.100md.com
[9]Jameel S,Zafrullah M,Ozdener MH,et al.Expression in animal cells and characterization of the hepatitis E virus structural proteins.J Virol,1996,70:207-216.
[10]Torresi J,Meanger J,Lambert P,et al.High level expression of the capsid protein of hepatitis E virus in diverse eukaryotic cells using the Semliki Forest virus replicon.J Virol Methods,1997,69:81-91.
[11]Torresi J,Li F,Locarnini SA,et al.Only the non-glycosylated frac-tion of hepatits E virus capsid (open reading frame 2) protein is stable in mammaliam cells.Gen Virol,1999,80(Pt5):1185-1188.
收稿日期：1999-08-11, http://www.100md.com
单位：100083 北京医科大学微生物系
关键词：
肝脏000132 戊型肝炎病毒(HEV)为线状单股正链RNA病毒，基因组全长约7.5kb(7.2～7.6kb)，编码2 400～2 533个氨基酸，由5’端非结构区(NS)和3’端结构区(S)组成，3’端带一个由150～200个腺苷酸残基组成的poly A尾巴。HEV基因组含3个开放读码框架(ORFs)，ORF1位于5’NS区，长5 079bp(第28～5107nt)，编码HEV复制相关蛋白，包括甲基转移酶、木瓜酶样的蛋白酶、解旋酶、RNA依赖性RNA聚合酶(RDRP)；ORF2从ORF1终止密码下游37个bp开始，长1 980bp(第5 147～ 7127nt)，编码660个氨基酸，主要包括信号肽和衣壳蛋白；ORF3位于ORF1、ORF2之间(第5 106～5 478nt)，与两者均有部分重叠，长369bp，编码一段123个氨基酸的多肽，功能不清。有学者认为ORF3编码的蛋白是一种与细胞骨架有关的磷蛋白，也有认为与转膜元件有关。本文仅就HEV抗原表位及ORF2抗原真核表达的研究进展简要综述如下。
, 百拇医药
一、HEV抗原表位的研究
(一)ORF1编码蛋白的抗原表位 Kaur等[1]在根据ORF1序列设计合成的系列多肽片段中，发现人类线状B细胞抗原表位存在于12个独立的多肽片段，其中有5个集中于第1207～1 446aa，该区域正处于RDRP内。RDRP内的抗原表位可能与非保护性反应有关；这些抗原也可能派生出附着于感染细胞膜表面的抗原位点，从而参与抗体依赖的抗病毒机制，如抗体依赖性细胞毒作用。……
参考文献：
[1]Kaur M,Hyams KC,Purdy MA,et al.Human linear B-cell epitopes encoded by the hepatitis E virus include determinants in the RNA-de-pendent RNA polymerase.Proc Natl Acad Sci USA,1992,89:3855-3858.
, 百拇医药
[2]Li F,Zhuang H,Kolivas S,et al.Persistent and transient antibody responses to hepatitis E virus detected by western immunoblot using open reading frame 2 and 3 and glutathione S-transferase fusion pro-teins.J Clin Microbiol,1994,32:2060-2066.
[3]Li F,Torresi J,Locarnini SA,et al.Amino-terminal epitopes are ex-posed when full-length open reading frame 2 of hepatits E virus is ex-pressed in Escherichia coil,but carboxyterrninal epitopes are masked.J Med Virol,1997,52:289-300.
, 百拇医药
[4]Purdy MA,McCaustland KA,Krawczynsk K,et al.Expression of a hepatitis E virus (HEV)-trpE fusion protein containing epitopes recognized by antibodies in sera from human cases and experimentally infected primates.Arch Virol,1992,123:335-349.
[5]Tsarev SA,Tsareva TS,Emerson SU,et al.ELISA for antibody to hepatitis E virus (HEV) based on complete open-reading frame-2 pro-tein expressed in insect cells:identification of HEV infection pri-mates.J Infect Dis,1993,168:369-378.
, 百拇医药
[6]Robinson RA,Burgess WH,Emerson SU,et al.Structural charac-terization of recombinant hepatitis E virus ORF2 proteins in bac-ulovirus-infected insect cells.Protein Exp Puri,1998,12:75-84.
[7]McAtee CP,Zhang YF,Yarbough PO,et al.Purification and characterization of a recombinant hepatits E protein vaccine candidate by liquid chromatography-mass spectrometry.J Chromatogr B Biomed Appl,1996,685:91-104.
[8]Li TC,Yamakawa Y,Suzuki K,et al.Expression and self-assembly of empty virus-like particles of hepatitis E virus.J Virol,1997,71:7207-7213.
, http://www.100md.com
[9]Jameel S,Zafrullah M,Ozdener MH,et al.Expression in animal cells and characterization of the hepatitis E virus structural proteins.J Virol,1996,70:207-216.
[10]Torresi J,Meanger J,Lambert P,et al.High level expression of the capsid protein of hepatitis E virus in diverse eukaryotic cells using the Semliki Forest virus replicon.J Virol Methods,1997,69:81-91.
[11]Torresi J,Li F,Locarnini SA,et al.Only the non-glycosylated frac-tion of hepatits E virus capsid (open reading frame 2) protein is stable in mammaliam cells.Gen Virol,1999,80(Pt5):1185-1188.
收稿日期：1999-08-11, http://www.100md.com