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【技术产业】新的丙型肝炎病毒培养方法

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http://www.eurekalert.org/pub_releases/2007-01/ajop-ncm011807.php

Public release date: 23-Jan-2007

New culture method for hepatitis C virus uses primary hepatocytes and patient serum
Seattle, WA -- Researchers open the way for improved study of hepatitis C virus by devising a novel virus culture system that allows replication of patient-isolated virus in nontransformed hepatocytes, instead of culture-adapted virus strains in transformed cell lines. The related report by L醶aro et al, "Hepatitis C virus replication in transfected and serum-infected cultured human fetal hepatocytes," appears in the February issue of The American Journal of Pathology.

Hepatitis C virus (HCV) infection affects approximately 170,000,000 people worldwide. HCV liver disease, which may induce liver inflammation, cirrhosis, and/or hepatocellular carcinoma, represents the foremost reason for liver transplantation in much of the U.S.

Study of HCV replication within liver cells, or hepatocytes, has been hampered by a lack of adequate virus culture systems. Some systems allow the virus to infect cells but do not permit prolonged replication and production of virus, while other systems rely on derivatives of permissive virus isolates for efficient replication in transformed (mutated) cell lines. Still lacking has been a system to sustain replication of novel virus isolates from patients using nontransformed hepatocytes.

Nelson Fausto of the University of Washington School of Medicine has crossed this hurdle using a human fetal hepatocyte culture system that was previously developed in his lab. Using this system, his group has demonstrated sustained replication and production of virus particles for at least 2 months, with these virus particles able to infect new cells.

In their first experiments, Fausto and colleagues transfected hepatocyte cultures with HCV genomic RNA and found replication of HCV RNA genomes and production of core protein (for virus particle formation). Release of infectious virus particles was confirmed, as media from these cells were able to infect naive hepatocytes. Finally, virus particles were examined by electron microscopy and shown to possess the expected size and shape of HCV virus particles.

Once the system was established, the group examined whether sera from patients carrying HCV could infect the human fetal hepatocytes. When sera from patients infected with different HCV strains were added to the hepatocyte culture system, viral replication occurred and new virus particles were produced.

In both transfection and infection models, virus particles were released in a cyclical manner, with bursts of virus produced every 10-14 days. This is similar to what has been reported during clinical HCV infection, possibly due to the host's natural defenses. Interestingly, cultured hepatocytes responded to viral replication by displaying signs of distress and cell death and by expressing interferon-beta, a cellular antiviral, in an effort to control the infection.

This culture system provides a breakthrough in studying HCV replication in nontransformed hepatocytes, the natural target of the virus. By allowing infection by patient serum containing a wide array of virus strains, this system may allow better understanding of the differences between different strains, further improving treatment strategies.

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2007-01-23 23:39 浏览 : 1075 回复 : 3

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本人已认领该文编译，48小时后若未提交译文，请其他战友自由认领。

2007-01-23 23:57

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Public release date: 23-Jan-2007
公开发行日期：2007-1-23
New culture method for hepatitis C virus uses primary hepatocytes and patient serum
利用胎儿肝脏细胞和患者血清进行新的丙型肝炎病毒培养方法
Seattle, WA -- Researchers open the way for improved study of hepatitis C virus by devising a novel virus culture system that allows replication of patient-isolated virus in nontransformed hepatocytes, instead of culture-adapted virus strains in transformed cell lines. The related report by L醶aro et al, "Hepatitis C virus replication in transfected and serum-infected cultured human fetal hepatocytes," appears in the February issue of The American Journal of Pathology.
西雅图WA--研究者已经发现HCV病毒新的培养方法，即利用非变异的肝细胞进行病毒复制，而不再采用肝变异细胞株。《美国病理学杂志》2月份期刊中L醶aro等相关报道说，“HCV病毒可在人类胎儿肝脏细胞转染复制”。
Hepatitis C virus (HCV) infection affects approximately 170,000,000 people worldwide. HCV liver disease, which may induce liver inflammation, cirrhosis, and/or hepatocellular carcinoma, represents the foremost reason for liver transplantation in much of the U.S.
全世界大约有1.7亿人口感染HCV。丙型肝炎可引起肝脏炎症、肝硬化和/或肝细胞癌，在美国是肝脏移植的最常见原因。
Study of HCV replication within liver cells, or hepatocytes, has been hampered by a lack of adequate virus culture systems. Some systems allow the virus to infect cells but do not permit prolonged replication and production of virus, while other systems rely on derivatives of permissive virus isolates for efficient replication in transformed (mutated) cell lines. Still lacking has been a system to sustain replication of novel virus isolates from patients using nontransformed hepatocytes.
有关HCV在肝脏细胞复制繁殖的很多研究，因为缺乏足够的病毒培养体系而被迫停止不能进行下去。一些培养体系可使病毒感染细胞而不能促进其大量复制繁殖，还有一些培养体系需要诱导剂使病毒在突变细胞株内充分复制。目前依然没有利用患者非突变肝细并能维持足够的病毒复制的培养体系。
Nelson Fausto of the University of Washington School of Medicine has crossed this hurdle using a human fetal hepatocyte culture system that was previously developed in his lab. Using this system, his group has demonstrated sustained replication and production of virus particles for at least 2 months, with these virus particles able to infect new cells.
华盛顿大学医学院Nelson Fausto首次在实验室利用人类胎儿肝细胞培养系统突破这些障碍。他们已经证实，采用这些培养系统可至少维持病毒颗粒复制繁殖2个月，并且这些病毒又能重新感染新的细胞。
In their first experiments, Fausto and colleagues transfected hepatocyte cultures with HCV genomic RNA and found replication of HCV RNA genomes and production of core protein (for virus particle formation). Release of infectious virus particles was confirmed, as media from these cells were able to infect naive hepatocytes. Finally, virus particles were examined by electron microscopy and shown to possess the expected size and shape of HCV virus particles.
在实验开始，Fausto和其同事把HCV的RNA转染给肝细胞，并观察到HCV RNA和核蛋白扩增复制。大量感染性病毒颗粒释放，然后又感染新生的肝细胞。最终，电镜观察到病毒颗粒形态结构均达到预想状态。
Once the system was established, the group examined whether sera from patients carrying HCV could infect the human fetal hepatocytes. When sera from patients infected with different HCV strains were added to the hepatocyte culture system, viral replication occurred and new virus particles were produced.
一旦该系统建立起了，研究小组又检验HCV携带者的血清能否感染人类胎儿肝细胞。发现当HCV携带者的血清加入该培养系统后，仍有大量病毒复制繁殖。
In both transfection and infection models, virus particles were released in a cyclical manner, with bursts of virus produced every 10-14 days. This is similar to what has been reported during clinical HCV infection, possibly due to the host's natural defenses. Interestingly, cultured hepatocytes responded to viral replication by displaying signs of distress and cell death and by expressing interferon-beta, a cellular antiviral, in an effort to control the infection.
上述两种转染和感染模型中，发现病毒颗粒呈周期性复制，一般10-14天增加一倍。这与临床HCV感染病例结果相似，可能和宿主细胞主动免疫有关。有趣的是，培养系统同时也通过应激或细胞凋亡来对抗病毒复制，并且表达β干扰素，产生细胞毒性，控制炎症进展。
This culture system provides a breakthrough in studying HCV replication in nontransformed hepatocytes, the natural target of the virus. By allowing infection by patient serum containing a wide array of virus strains, this system may allow better understanding of the differences between different strains, further improving treatment strategies.
该病毒培养系统是非突变肝细胞内HCV复制扩增技术的突破。通过患者的感染血清加入培养系统，使得我们更好的认识不同感染机制的差别，并能指引改进治疗策略。

编译：标题：利用胎儿肝脏细胞和患者血清进行新的丙型肝炎病毒培养方法

利用胎儿肝脏细胞和患者血清进行新的丙型肝炎病毒培养方法

西雅图WA--研究者已经发现HCV病毒新的培养方法，即利用非变异的肝细胞进行病毒复制，而不再采用肝变异细胞株。《美国病理学杂志》2月份期刊中L醶aro等相关报道说，“HCV病毒可在转染或加入感染血清的人类胎儿肝脏细胞中复制”。

全世界大约有1.7亿人口感染HCV。丙型肝炎可引起肝脏炎症、肝硬化和/或肝细胞癌，在美国是肝脏移植的最常见原因。

有关HCV在肝脏细胞复制繁殖的很多研究，因为缺乏足够的病毒培养体系而被迫停止不能进行下去。一些培养体系可使病毒感染细胞而不能促进其大量复制繁殖，还有一些培养体系需要诱导剂使病毒在突变细胞株内充分复制。目前依然没有利用患者非突变肝细并能维持足够的病毒复制的培养体系。

华盛顿大学医学院Nelson Fausto首次在实验室利用人类胎儿肝细胞培养系统突破这些障碍。他们已经证实，采用这些培养系统可至少维持病毒颗粒复制繁殖2个月，并且这些病毒又能重新感染新的细胞。在实验开始，Fausto和其同事把HCV的RNA转染给肝细胞，并观察到HCV RNA和核蛋白扩增复制。大量感染性病毒颗粒释放，然后又感染新生的肝细胞。最终，电镜观察到病毒颗粒形态结构均达到预想状态。一旦该系统建立起了，研究小组又检验HCV携带者的血清能否感染人类胎儿肝细胞。发现当HCV携带者的血清加入该培养系统后，仍有大量病毒复制繁殖。

上述两种转染和感染模型中，发现病毒颗粒呈周期性复制，一般10-14天增加一倍。这与临床HCV感染病例结果相似，可能和宿主细胞主动免疫有关。有趣的是，培养系统同时也通过应激或细胞凋亡来对抗病毒复制，并且表达β干扰素，产生细胞毒性，控制炎症进展。

该病毒培养系统是非突变肝细胞内HCV复制扩增技术的突破。通过患者的感染血清加入培养系统，使得我们更好的认识不同感染机制的差别，并能指引改进治疗策略。

2007-01-24 22:33

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请问culture-adapted virus是什么？谢谢

2018-03-02 10:58 来自 Android客户端

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• # 妇儿联动 # 启动期，作为妇产科/儿科医生，你有哪些问题，想请教对方？

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