Science:攻克长达48年的难题,首次在人类肠道细胞培养诺如病毒

2016-08-29 佚名 生物探索

1968年,科学家首次从急性腹泻患者粪便分离出来人类诺如病毒,后者全球各地引起急性泻的主要病原体。遗憾的是,该病毒不能在小鼠或其他模型上进行生长和研究。为此,科学家通过长达48年的探索,终于首次在人类肠道细胞培养出了诺如病毒。该研究近日发表在《科学》(Science)上。

1968年,科学家首次从急性腹泻患者粪便分离出来人类诺如病毒,后者全球各地引起急性腹泻的主要病原体。遗憾的是,该病毒不能在小鼠或其他模型上进行生长和研究。为此,科学家通过长达48年的探索,终于首次在人类肠道细胞培养出了诺如病毒。该研究近日发表在《科学》(Science)上。

诺如病毒——急性腹泻的主要诱因

人类诺如病毒(Human noroviruses),是全球各地引起急性腹泻的主要病原体,全年均可发生感染,但每年10月到次年3月是暴发流行的高发季,也被称为“冬季呕吐病”。感染者一般在摄入病毒后24-48小时出现症状,发病突然,主要症状为恶心、呕吐、腹部痉挛性腹泻等。

据统计,美国每年的非细菌性腹泻有60-90%是由诺如病毒引起;荷兰、英国、日本、澳大利亚等发达国家情况不相上下;中国5岁以下腹泻儿童中,诺如病毒检出率为15%左右,血清抗体水平调查表明中国人群中诺如病毒的感染亦十分普遍。

尽管“诺如病毒”感染性腹泻的情况非常普遍,但是目前仍然没有疫苗和特效药物。此外,关于“诺如病毒”的机制很难研究,因为自Albert Kapikian教授在1968年首次从急性腹泻患者粪便分离出来后,科学家一直未找到合适的方式在实验室里培养该病毒。

人类诺如病毒:高度的物种专一性,难以培育

诺如病毒,又称诺瓦克病毒(Norwalk Viruses,NV),具有高度的物种专一性,即人类诺如病毒是只感染人类,鼠类诺如病毒只感染鼠类。因此,人类诺如病毒不能在小鼠或其他模型上进行生长和研究。

因此,人类诺如病毒无法感染生物医学研究中常用的任何物种,如小鼠、大鼠或兔子。人类诺如病毒甚至不能感染培养皿中的人类细胞。

首次在人类肠道细胞培养诺如病毒

近日,贝勒医学院的科学家们终于攻克了这个困扰人类40多年的难题,首次在实验室里的人类肠道上皮细胞上培养了诺如病毒,并将这项成果发表在8月25日的《科学》(Science)上。这意味着人类向胃肠炎病毒的研究迈向了重要的一步。

贝勒医学院分子病毒学教授、文章的资深作者Mary Estes表示:数十年来,人类一直尝试在实验室里培育诺如病毒,其所在实验室也尝试了20多年,尽管在培育的过程中找到了很多其他病毒,但是一直很难找到诺如病毒的工作机制。

为构建适合人类诺如病毒的细胞花了5年时间

经过多年的探索,Estes教授发现之所以培育不了人类诺如病毒,是因为找不到合适的细胞类型。后来,他们首次在慢性诺如病毒感染的患者身上发现:该病毒可以在肠上皮细胞上被检测到,但是在正常的肠上皮细胞上培育的病毒会迅速死亡。

荷兰的Hans Clevers团队曾开发出了一种全新的包括肠上皮细胞的人类肠道上皮细胞培养系统,这种新型的、多细胞人源化培养系统叫做“enteroids”,是从患者组织里提取的成年的肠道干细胞。于是Estes教授团队经过5年多时间的探索,在尝试了众多方法失败后,他们开始使用全新的经肠道分离的人体干细胞培育。一年后,他们终于在实验室里获得了生长良好的人肠上皮细胞。

胆汁,是最适合人类诺如病毒生长的培养基

而后,在他们测试诺如病毒之前,他们首先测试了另外一种人类胃肠病毒——轮状病毒(rotavirus),发现轮状病毒在人肠上皮细胞中培育的非常好。

在尝试人类诺如病毒时,却发现仅有少数菌株生长可以,其他的均不能存活。于是,他们加入了一些其他自然存在于小肠上部的培养物质。

众所周知,人体对食物的反应是由胰腺和胆汁分泌的酶从肝脏分泌到小肠内的。胰腺酶消化大分子和胆汁来溶解脂肪。轮状病毒利用胰腺酶触发其复制来引起的胃肠炎;但这些酶对诺如病毒并没有影响。

因此,科学家猜测,既然酶不起作用,那么胆汁是否会成为允许病毒的复制的基质呢?于是,科学家加入了胆汁,并发现了之前未生长的菌株都存活了。

科学家们为他们的发现而感到震惊,原来胆汁是存进人类诺如病毒复制的“催化剂”。

研究意义:进一步探索预防和治疗感染的途径

就在这项研究发表的前几天,华盛顿大学和Broad研究所的科学家小组在CRISPR/Cas9技术的帮助下,找到对小鼠诺如病毒感染至关重要的基因。但是小鼠诺如病毒并不能感染人类,也难以成为研究药物或者疫苗的方法。

该项突破建立了一个人类诺如病毒可以生长的生态系统,有助于研究人员进一步探索预防和治疗感染的途径,并更好地理解诺如病毒的生物学机制。

过去无法在实验室里培养的人类诺如病毒,将在这个全新的系统中进行培育,提高了科学家未来开发抗病毒药物的能力。

原始出处:
Ettayebi K1, Crawford SE1, Murakami K1, Broughman JR1, Karandikar U1, Tenge VR1, Neill FH1, Blutt SE1, Zeng XL1, Qu L1, Kou B1, Opekun AR2, Burrin D3, Graham DY4, Ramani S1, Atmar RL5, Estes MK6.Replication of human noroviruses in stem cell-derived human enteroids.Science. 2016 Aug 25 

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createdTime=Mon Aug 29 14:54:00 CST 2016, time=2016-08-29, status=1, ipAttribution=)]
    2016-10-12 1e0d99ddm02(暂无匿称)

    总结的很好!

    0

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createdTime=Mon Aug 29 14:54:00 CST 2016, time=2016-08-29, status=1, ipAttribution=)]
    2016-10-12 1e0d99ddm02(暂无匿称)

    分享一下!

    0

  3. 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createdTime=Mon Aug 29 14:54:00 CST 2016, time=2016-08-29, status=1, ipAttribution=)]
    2016-09-23 1e10c84am36(暂无匿称)

    文章不错,继续学习

    0

  4. 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createdName=jichang, createdTime=Wed Aug 31 04:26:00 CST 2016, time=2016-08-31, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1479195, encodeId=d97114e919546, content=<a href='/topic/show?id=82d0919287c' target=_blank style='color:#2F92EE;'>#诺如病毒#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=27, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=91928, encryptionId=82d0919287c, topicName=诺如病毒)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=d8627501386, createdName=丁鹏鹏, createdTime=Wed Aug 31 04:26:00 CST 2016, time=2016-08-31, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=106907, encodeId=49c710690e4d, content=学习了解一下, beContent=null, objectType=article, channel=null, level=null, likeNumber=18, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=90f31612316, createdName=xiaotaiyang1, createdTime=Mon Aug 29 14:54:00 CST 2016, time=2016-08-29, status=1, ipAttribution=)]
    2016-09-13 lhl629

    学习学习。。。

    0

  5. 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createdName=jichang, createdTime=Wed Aug 31 04:26:00 CST 2016, time=2016-08-31, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1479195, encodeId=d97114e919546, content=<a href='/topic/show?id=82d0919287c' target=_blank style='color:#2F92EE;'>#诺如病毒#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=27, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=91928, encryptionId=82d0919287c, topicName=诺如病毒)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=d8627501386, createdName=丁鹏鹏, createdTime=Wed Aug 31 04:26:00 CST 2016, time=2016-08-31, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=106907, encodeId=49c710690e4d, content=学习了解一下, beContent=null, objectType=article, channel=null, level=null, likeNumber=18, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=90f31612316, createdName=xiaotaiyang1, createdTime=Mon Aug 29 14:54:00 CST 2016, time=2016-08-29, status=1, ipAttribution=)]
    2016-09-10 1def6e10m76(暂无匿称)

    很好的课程,学到很多,继续学习

    0

  6. 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createdName=jichang, createdTime=Wed Aug 31 04:26:00 CST 2016, time=2016-08-31, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1479195, encodeId=d97114e919546, content=<a href='/topic/show?id=82d0919287c' target=_blank style='color:#2F92EE;'>#诺如病毒#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=27, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=91928, encryptionId=82d0919287c, topicName=诺如病毒)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=d8627501386, createdName=丁鹏鹏, createdTime=Wed Aug 31 04:26:00 CST 2016, time=2016-08-31, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=106907, encodeId=49c710690e4d, content=学习了解一下, beContent=null, objectType=article, channel=null, level=null, likeNumber=18, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=90f31612316, createdName=xiaotaiyang1, createdTime=Mon Aug 29 14:54:00 CST 2016, time=2016-08-29, status=1, ipAttribution=)]
    2016-09-08 lhl629

    很好的内容

    0

  7. 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createdName=jichang, createdTime=Wed Aug 31 04:26:00 CST 2016, time=2016-08-31, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1479195, encodeId=d97114e919546, content=<a href='/topic/show?id=82d0919287c' target=_blank style='color:#2F92EE;'>#诺如病毒#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=27, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=91928, encryptionId=82d0919287c, topicName=诺如病毒)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=d8627501386, createdName=丁鹏鹏, createdTime=Wed Aug 31 04:26:00 CST 2016, time=2016-08-31, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=106907, encodeId=49c710690e4d, content=学习了解一下, beContent=null, objectType=article, channel=null, level=null, likeNumber=18, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=90f31612316, createdName=xiaotaiyang1, createdTime=Mon Aug 29 14:54:00 CST 2016, time=2016-08-29, status=1, ipAttribution=)]
    2016-09-05 李东泽

    很好,不错,以后会多学习

    0

  8. 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GetPortalCommentsPageByObjectIdResponse(id=111224, encodeId=f6a9111224f4, content=很好,不错,以后会多学习, beContent=null, objectType=article, channel=null, level=null, likeNumber=23, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=https://wx.qlogo.cn/mmopen/jW482SpianMayicTRbRZ5RzUn81b5CF5ibVPib8jWI92iciaxmqGHlwruOnK4SVZykd4iahqo1hicklIibyZoxm55X7EM8BW9eX1h1hQ2/0, createdBy=8aa81937526, createdName=李东泽, createdTime=Mon Sep 05 10:10:00 CST 2016, time=2016-09-05, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1375323, encodeId=dec713e5323d3, content=<a href='/topic/show?id=2949159989d' target=_blank style='color:#2F92EE;'>#SCIE#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=24, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=15998, encryptionId=2949159989d, topicName=SCIE)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=17a3387, createdName=jichang, createdTime=Wed Aug 31 04:26:00 CST 2016, time=2016-08-31, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1479195, encodeId=d97114e919546, content=<a href='/topic/show?id=82d0919287c' target=_blank style='color:#2F92EE;'>#诺如病毒#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=27, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=91928, encryptionId=82d0919287c, topicName=诺如病毒)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=d8627501386, createdName=丁鹏鹏, createdTime=Wed Aug 31 04:26:00 CST 2016, time=2016-08-31, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=106907, encodeId=49c710690e4d, content=学习了解一下, beContent=null, objectType=article, channel=null, level=null, likeNumber=18, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=90f31612316, createdName=xiaotaiyang1, createdTime=Mon Aug 29 14:54:00 CST 2016, time=2016-08-29, status=1, ipAttribution=)]
    2016-08-31 jichang
  9. 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createdTime=Mon Aug 29 14:54:00 CST 2016, time=2016-08-29, status=1, ipAttribution=)]
  10. 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createdTime=Mon Aug 29 14:54:00 CST 2016, time=2016-08-29, status=1, ipAttribution=)]
    2016-08-29 xiaotaiyang1

    学习了解一下

    0

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