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17th Century Variola Virus Reveals the Recent History of Smallpox

Identifieur interne : 001098 ( Pmc/Curation ); précédent : 001097; suivant : 001099

17th Century Variola Virus Reveals the Recent History of Smallpox

Auteurs : Ana T. Duggan [Canada] ; Maria F. Perdomo [Finlande] ; Dario Piombino-Mascali [Lituanie] ; Stephanie Marciniak [Canada] ; Debi Poinar [Canada] ; Matthew V. Emery [Canada] ; Jan P. Buchmann [Australie] ; Sebastian Duchêne [Australie] ; Rimantas Jankauskas [Lituanie] ; Margaret Humphreys [États-Unis] ; G. Brian Golding [Canada] ; John Southon [États-Unis] ; Alison Devault [États-Unis] ; Jean-Marie Rouillard [États-Unis] ; Jason W. Sahl [États-Unis] ; Olivier Dutour [France] ; Klaus Hedman [Finlande] ; Antti Sajantila [Finlande] ; Geoffrey L. Smith [Royaume-Uni] ; Edward C. Holmes [Australie] ; Hendrik N. Poinar [Canada]

Source :

RBID : PMC:5196022

Abstract

Summary

Smallpox holds a unique position in the history of medicine. It was the first disease for which a vaccine was developed and remains the only human disease eradicated by vaccination. Although there have been claims of smallpox in Egypt, India, and China dating back millennia [1, 2, 3, 4], the timescale of emergence of the causative agent, variola virus (VARV), and how it evolved in the context of increasingly widespread immunization, have proven controversial [4, 5, 6, 7, 8, 9]. In particular, some molecular-clock-based studies have suggested that key events in VARV evolution only occurred during the last two centuries [4, 5, 6] and hence in apparent conflict with anecdotal historical reports, although it is difficult to distinguish smallpox from other pustular rashes by description alone. To address these issues, we captured, sequenced, and reconstructed a draft genome of an ancient strain of VARV, sampled from a Lithuanian child mummy dating between 1643 and 1665 and close to the time of several documented European epidemics [1, 2, 10]. When compared to vaccinia virus, this archival strain contained the same pattern of gene degradation as 20th century VARVs, indicating that such loss of gene function had occurred before ca. 1650. Strikingly, the mummy sequence fell basal to all currently sequenced strains of VARV on phylogenetic trees. Molecular-clock analyses revealed a strong clock-like structure and that the timescale of smallpox evolution is more recent than often supposed, with the diversification of major viral lineages only occurring within the 18th and 19th centuries, concomitant with the development of modern vaccination.


Url:
DOI: 10.1016/j.cub.2016.10.061
PubMed: 27939314
PubMed Central: 5196022

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PMC:5196022

Le document en format XML

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<sup>th</sup>
Century Variola Virus Reveals the Recent History of Smallpox</title>
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<name sortKey="Duggan, Ana T" sort="Duggan, Ana T" uniqKey="Duggan A" first="Ana T." last="Duggan">Ana T. Duggan</name>
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<nlm:aff id="aff1">McMaster Ancient DNA Centre, Department of Anthropology, McMaster University, Hamilton, ON L8S 4L8, Canada</nlm:aff>
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<name sortKey="Perdomo, Maria F" sort="Perdomo, Maria F" uniqKey="Perdomo M" first="Maria F." last="Perdomo">Maria F. Perdomo</name>
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<name sortKey="Piombino Mascali, Dario" sort="Piombino Mascali, Dario" uniqKey="Piombino Mascali D" first="Dario" last="Piombino-Mascali">Dario Piombino-Mascali</name>
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<name sortKey="Marciniak, Stephanie" sort="Marciniak, Stephanie" uniqKey="Marciniak S" first="Stephanie" last="Marciniak">Stephanie Marciniak</name>
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<name sortKey="Emery, Matthew V" sort="Emery, Matthew V" uniqKey="Emery M" first="Matthew V." last="Emery">Matthew V. Emery</name>
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<name sortKey="Buchmann, Jan P" sort="Buchmann, Jan P" uniqKey="Buchmann J" first="Jan P." last="Buchmann">Jan P. Buchmann</name>
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<name sortKey="Duchene, Sebastian" sort="Duchene, Sebastian" uniqKey="Duchene S" first="Sebastian" last="Duchêne">Sebastian Duchêne</name>
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<nlm:aff id="aff5">Department of Biochemistry and Molecular Biology, Bio21 Molecular Science and Biotechnology Institute, The University of Melbourne, Melbourne, VIC 3010, Australia</nlm:aff>
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<name sortKey="Jankauskas, Rimantas" sort="Jankauskas, Rimantas" uniqKey="Jankauskas R" first="Rimantas" last="Jankauskas">Rimantas Jankauskas</name>
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<name sortKey="Golding, G Brian" sort="Golding, G Brian" uniqKey="Golding G" first="G. Brian" last="Golding">G. Brian Golding</name>
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<name sortKey="Southon, John" sort="Southon, John" uniqKey="Southon J" first="John" last="Southon">John Southon</name>
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<name sortKey="Devault, Alison" sort="Devault, Alison" uniqKey="Devault A" first="Alison" last="Devault">Alison Devault</name>
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<name sortKey="Rouillard, Jean Marie" sort="Rouillard, Jean Marie" uniqKey="Rouillard J" first="Jean-Marie" last="Rouillard">Jean-Marie Rouillard</name>
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<nlm:aff id="aff10">Department of Chemical Engineering, University of Michigan, Ann Arbor, MI 48109-2136, USA</nlm:aff>
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<wicri:regionArea>Department of Chemical Engineering, University of Michigan, Ann Arbor, MI 48109-2136</wicri:regionArea>
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<name sortKey="Sahl, Jason W" sort="Sahl, Jason W" uniqKey="Sahl J" first="Jason W." last="Sahl">Jason W. Sahl</name>
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<nlm:aff id="aff11">Center for Microbial Genetics and Genomics, Northern Arizona University, Flagstaff, AZ 86011-4073, USA</nlm:aff>
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<wicri:regionArea>Center for Microbial Genetics and Genomics, Northern Arizona University, Flagstaff, AZ 86011-4073</wicri:regionArea>
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<name sortKey="Dutour, Olivier" sort="Dutour, Olivier" uniqKey="Dutour O" first="Olivier" last="Dutour">Olivier Dutour</name>
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<nlm:aff id="aff12">Laboratoire d’Anthropologie Biologique Paul Broca, Ecole Pratique des Hautes Etudes, PSL Research University, Paris 75014, France</nlm:aff>
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<nlm:aff id="aff13">PACEA, CNRS, Université de Bordeaux, Pessac 33615, France</nlm:aff>
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<name sortKey="Hedman, Klaus" sort="Hedman, Klaus" uniqKey="Hedman K" first="Klaus" last="Hedman">Klaus Hedman</name>
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<nlm:aff id="aff2">Department of Virology, University of Helsinki, Helsinki 00014, Finland</nlm:aff>
<country xml:lang="fr">Finlande</country>
<wicri:regionArea>Department of Virology, University of Helsinki, Helsinki 00014</wicri:regionArea>
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<nlm:aff id="aff14">Helsinki University Hospital, Helsinki 00029, Finland</nlm:aff>
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<name sortKey="Sajantila, Antti" sort="Sajantila, Antti" uniqKey="Sajantila A" first="Antti" last="Sajantila">Antti Sajantila</name>
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<nlm:aff id="aff15">Department of Forensic Medicine, University of Helsinki, Helsinki 00014, Finland</nlm:aff>
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<name sortKey="Smith, Geoffrey L" sort="Smith, Geoffrey L" uniqKey="Smith G" first="Geoffrey L." last="Smith">Geoffrey L. Smith</name>
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<nlm:aff id="aff16">Department of Pathology, University of Cambridge, Cambridge CB2 1QP, UK</nlm:aff>
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<name sortKey="Holmes, Edward C" sort="Holmes, Edward C" uniqKey="Holmes E" first="Edward C." last="Holmes">Edward C. Holmes</name>
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<name sortKey="Poinar, Hendrik N" sort="Poinar, Hendrik N" uniqKey="Poinar H" first="Hendrik N." last="Poinar">Hendrik N. Poinar</name>
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<nlm:aff id="aff17">Michael G. DeGroote Institute for Infectious Disease Research, McMaster University, Hamilton, ON L8S 4L8, Canada</nlm:aff>
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<wicri:regionArea>Michael G. DeGroote Institute for Infectious Disease Research, McMaster University, Hamilton, ON L8S 4L8</wicri:regionArea>
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<nlm:aff id="aff18">Humans and the Microbiome Program, Canadian Institute for Advanced Research, Toronto, ON M5G 1Z8, Canada</nlm:aff>
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<title xml:lang="en" level="a" type="main">17
<sup>th</sup>
Century Variola Virus Reveals the Recent History of Smallpox</title>
<author>
<name sortKey="Duggan, Ana T" sort="Duggan, Ana T" uniqKey="Duggan A" first="Ana T." last="Duggan">Ana T. Duggan</name>
<affiliation wicri:level="1">
<nlm:aff id="aff1">McMaster Ancient DNA Centre, Department of Anthropology, McMaster University, Hamilton, ON L8S 4L8, Canada</nlm:aff>
<country xml:lang="fr">Canada</country>
<wicri:regionArea>McMaster Ancient DNA Centre, Department of Anthropology, McMaster University, Hamilton, ON L8S 4L8</wicri:regionArea>
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<name sortKey="Perdomo, Maria F" sort="Perdomo, Maria F" uniqKey="Perdomo M" first="Maria F." last="Perdomo">Maria F. Perdomo</name>
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<nlm:aff id="aff2">Department of Virology, University of Helsinki, Helsinki 00014, Finland</nlm:aff>
<country xml:lang="fr">Finlande</country>
<wicri:regionArea>Department of Virology, University of Helsinki, Helsinki 00014</wicri:regionArea>
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<name sortKey="Piombino Mascali, Dario" sort="Piombino Mascali, Dario" uniqKey="Piombino Mascali D" first="Dario" last="Piombino-Mascali">Dario Piombino-Mascali</name>
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<nlm:aff id="aff3">Department of Anatomy, Histology, and Anthropology, Faculty of Medicine, Vilnius University, Vilnius 03101, Lithuania</nlm:aff>
<country xml:lang="fr">Lituanie</country>
<wicri:regionArea>Department of Anatomy, Histology, and Anthropology, Faculty of Medicine, Vilnius University, Vilnius 03101</wicri:regionArea>
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<name sortKey="Marciniak, Stephanie" sort="Marciniak, Stephanie" uniqKey="Marciniak S" first="Stephanie" last="Marciniak">Stephanie Marciniak</name>
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<nlm:aff id="aff1">McMaster Ancient DNA Centre, Department of Anthropology, McMaster University, Hamilton, ON L8S 4L8, Canada</nlm:aff>
<country xml:lang="fr">Canada</country>
<wicri:regionArea>McMaster Ancient DNA Centre, Department of Anthropology, McMaster University, Hamilton, ON L8S 4L8</wicri:regionArea>
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<name sortKey="Poinar, Debi" sort="Poinar, Debi" uniqKey="Poinar D" first="Debi" last="Poinar">Debi Poinar</name>
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<nlm:aff id="aff1">McMaster Ancient DNA Centre, Department of Anthropology, McMaster University, Hamilton, ON L8S 4L8, Canada</nlm:aff>
<country xml:lang="fr">Canada</country>
<wicri:regionArea>McMaster Ancient DNA Centre, Department of Anthropology, McMaster University, Hamilton, ON L8S 4L8</wicri:regionArea>
</affiliation>
</author>
<author>
<name sortKey="Emery, Matthew V" sort="Emery, Matthew V" uniqKey="Emery M" first="Matthew V." last="Emery">Matthew V. Emery</name>
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<nlm:aff id="aff1">McMaster Ancient DNA Centre, Department of Anthropology, McMaster University, Hamilton, ON L8S 4L8, Canada</nlm:aff>
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<name sortKey="Buchmann, Jan P" sort="Buchmann, Jan P" uniqKey="Buchmann J" first="Jan P." last="Buchmann">Jan P. Buchmann</name>
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<nlm:aff id="aff4">Marie Bashir Institute for Infectious Diseases and Biosecurity, Charles Perkins Centre, School of Life and Environmental Sciences and Sydney Medical School, The University of Sydney, Sydney, NSW 2145, Australia</nlm:aff>
<country xml:lang="fr">Australie</country>
<wicri:regionArea>Marie Bashir Institute for Infectious Diseases and Biosecurity, Charles Perkins Centre, School of Life and Environmental Sciences and Sydney Medical School, The University of Sydney, Sydney, NSW 2145</wicri:regionArea>
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<name sortKey="Duchene, Sebastian" sort="Duchene, Sebastian" uniqKey="Duchene S" first="Sebastian" last="Duchêne">Sebastian Duchêne</name>
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<nlm:aff id="aff4">Marie Bashir Institute for Infectious Diseases and Biosecurity, Charles Perkins Centre, School of Life and Environmental Sciences and Sydney Medical School, The University of Sydney, Sydney, NSW 2145, Australia</nlm:aff>
<country xml:lang="fr">Australie</country>
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<nlm:aff id="aff5">Department of Biochemistry and Molecular Biology, Bio21 Molecular Science and Biotechnology Institute, The University of Melbourne, Melbourne, VIC 3010, Australia</nlm:aff>
<country xml:lang="fr">Australie</country>
<wicri:regionArea>Department of Biochemistry and Molecular Biology, Bio21 Molecular Science and Biotechnology Institute, The University of Melbourne, Melbourne, VIC 3010</wicri:regionArea>
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<name sortKey="Jankauskas, Rimantas" sort="Jankauskas, Rimantas" uniqKey="Jankauskas R" first="Rimantas" last="Jankauskas">Rimantas Jankauskas</name>
<affiliation wicri:level="1">
<nlm:aff id="aff3">Department of Anatomy, Histology, and Anthropology, Faculty of Medicine, Vilnius University, Vilnius 03101, Lithuania</nlm:aff>
<country xml:lang="fr">Lituanie</country>
<wicri:regionArea>Department of Anatomy, Histology, and Anthropology, Faculty of Medicine, Vilnius University, Vilnius 03101</wicri:regionArea>
</affiliation>
</author>
<author>
<name sortKey="Humphreys, Margaret" sort="Humphreys, Margaret" uniqKey="Humphreys M" first="Margaret" last="Humphreys">Margaret Humphreys</name>
<affiliation wicri:level="1">
<nlm:aff id="aff6">Department of History, Duke University, Durham, NC 27708-0719, USA</nlm:aff>
<country xml:lang="fr">États-Unis</country>
<wicri:regionArea>Department of History, Duke University, Durham, NC 27708-0719</wicri:regionArea>
</affiliation>
</author>
<author>
<name sortKey="Golding, G Brian" sort="Golding, G Brian" uniqKey="Golding G" first="G. Brian" last="Golding">G. Brian Golding</name>
<affiliation wicri:level="1">
<nlm:aff id="aff7">Department of Biology, McMaster University, Hamilton, ON L8S 4L8, Canada</nlm:aff>
<country xml:lang="fr">Canada</country>
<wicri:regionArea>Department of Biology, McMaster University, Hamilton, ON L8S 4L8</wicri:regionArea>
</affiliation>
</author>
<author>
<name sortKey="Southon, John" sort="Southon, John" uniqKey="Southon J" first="John" last="Southon">John Southon</name>
<affiliation wicri:level="1">
<nlm:aff id="aff8">Keck Carbon Cycle Accelerator Mass Spectrometer, Earth Systems Science Department, University of California, Irvine, CA 92697-3100, USA</nlm:aff>
<country xml:lang="fr">États-Unis</country>
<wicri:regionArea>Keck Carbon Cycle Accelerator Mass Spectrometer, Earth Systems Science Department, University of California, Irvine, CA 92697-3100</wicri:regionArea>
</affiliation>
</author>
<author>
<name sortKey="Devault, Alison" sort="Devault, Alison" uniqKey="Devault A" first="Alison" last="Devault">Alison Devault</name>
<affiliation wicri:level="1">
<nlm:aff id="aff9">MYcroarray, Ann Arbor, MI 48105, USA</nlm:aff>
<country xml:lang="fr">États-Unis</country>
<wicri:regionArea>MYcroarray, Ann Arbor, MI 48105</wicri:regionArea>
</affiliation>
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<nlm:aff id="aff9">MYcroarray, Ann Arbor, MI 48105, USA</nlm:aff>
<country xml:lang="fr">États-Unis</country>
<wicri:regionArea>MYcroarray, Ann Arbor, MI 48105</wicri:regionArea>
</affiliation>
<affiliation wicri:level="1">
<nlm:aff id="aff10">Department of Chemical Engineering, University of Michigan, Ann Arbor, MI 48109-2136, USA</nlm:aff>
<country xml:lang="fr">États-Unis</country>
<wicri:regionArea>Department of Chemical Engineering, University of Michigan, Ann Arbor, MI 48109-2136</wicri:regionArea>
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<nlm:aff id="aff11">Center for Microbial Genetics and Genomics, Northern Arizona University, Flagstaff, AZ 86011-4073, USA</nlm:aff>
<country xml:lang="fr">États-Unis</country>
<wicri:regionArea>Center for Microbial Genetics and Genomics, Northern Arizona University, Flagstaff, AZ 86011-4073</wicri:regionArea>
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<country xml:lang="fr">France</country>
<wicri:regionArea>Laboratoire d’Anthropologie Biologique Paul Broca, Ecole Pratique des Hautes Etudes, PSL Research University, Paris 75014</wicri:regionArea>
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<nlm:aff id="aff13">PACEA, CNRS, Université de Bordeaux, Pessac 33615, France</nlm:aff>
<country xml:lang="fr">France</country>
<wicri:regionArea>PACEA, CNRS, Université de Bordeaux, Pessac 33615</wicri:regionArea>
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<country xml:lang="fr">Finlande</country>
<wicri:regionArea>Department of Virology, University of Helsinki, Helsinki 00014</wicri:regionArea>
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<affiliation wicri:level="1">
<nlm:aff id="aff14">Helsinki University Hospital, Helsinki 00029, Finland</nlm:aff>
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<country xml:lang="fr">Finlande</country>
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<nlm:aff id="aff16">Department of Pathology, University of Cambridge, Cambridge CB2 1QP, UK</nlm:aff>
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<country xml:lang="fr">Australie</country>
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<nlm:aff id="aff7">Department of Biology, McMaster University, Hamilton, ON L8S 4L8, Canada</nlm:aff>
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<series>
<title level="j">Current Biology</title>
<idno type="ISSN">0960-9822</idno>
<idno type="eISSN">1879-0445</idno>
<imprint>
<date when="2016">2016</date>
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<textClass></textClass>
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<front>
<div type="abstract" xml:lang="en">
<title>Summary</title>
<p>Smallpox holds a unique position in the history of medicine. It was the first disease for which a vaccine was developed and remains the only human disease eradicated by vaccination. Although there have been claims of smallpox in Egypt, India, and China dating back millennia [
<xref rid="bib1" ref-type="bibr">1</xref>
,
<xref rid="bib2" ref-type="bibr">2</xref>
,
<xref rid="bib3" ref-type="bibr">3</xref>
,
<xref rid="bib4" ref-type="bibr">4</xref>
], the timescale of emergence of the causative agent, variola virus (VARV), and how it evolved in the context of increasingly widespread immunization, have proven controversial [
<xref rid="bib4" ref-type="bibr">4</xref>
,
<xref rid="bib5" ref-type="bibr">5</xref>
,
<xref rid="bib6" ref-type="bibr">6</xref>
,
<xref rid="bib7" ref-type="bibr">7</xref>
,
<xref rid="bib8" ref-type="bibr">8</xref>
,
<xref rid="bib9" ref-type="bibr">9</xref>
]. In particular, some molecular-clock-based studies have suggested that key events in VARV evolution only occurred during the last two centuries [
<xref rid="bib4" ref-type="bibr">4</xref>
,
<xref rid="bib5" ref-type="bibr">5</xref>
,
<xref rid="bib6" ref-type="bibr">6</xref>
] and hence in apparent conflict with anecdotal historical reports, although it is difficult to distinguish smallpox from other pustular rashes by description alone. To address these issues, we captured, sequenced, and reconstructed a draft genome of an ancient strain of VARV, sampled from a Lithuanian child mummy dating between 1643 and 1665 and close to the time of several documented European epidemics [
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,
<xref rid="bib2" ref-type="bibr">2</xref>
,
<xref rid="bib10" ref-type="bibr">10</xref>
]. When compared to vaccinia virus, this archival strain contained the same pattern of gene degradation as 20
<sup>th</sup>
century VARVs, indicating that such loss of gene function had occurred before ca. 1650. Strikingly, the mummy sequence fell basal to all currently sequenced strains of VARV on phylogenetic trees. Molecular-clock analyses revealed a strong clock-like structure and that the timescale of smallpox evolution is more recent than often supposed, with the diversification of major viral lineages only occurring within the 18
<sup>th</sup>
and 19
<sup>th</sup>
centuries, concomitant with the development of modern vaccination.</p>
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</biblStruct>
</listBibl>
</div1>
</back>
</TEI>
<pmc article-type="brief-report">
<pmc-dir>properties open_access</pmc-dir>
<front>
<journal-meta>
<journal-id journal-id-type="nlm-ta">Curr Biol</journal-id>
<journal-id journal-id-type="iso-abbrev">Curr. Biol</journal-id>
<journal-title-group>
<journal-title>Current Biology</journal-title>
</journal-title-group>
<issn pub-type="ppub">0960-9822</issn>
<issn pub-type="epub">1879-0445</issn>
<publisher>
<publisher-name>Cell Press</publisher-name>
</publisher>
</journal-meta>
<article-meta>
<article-id pub-id-type="pmid">27939314</article-id>
<article-id pub-id-type="pmc">5196022</article-id>
<article-id pub-id-type="publisher-id">S0960-9822(16)31324-0</article-id>
<article-id pub-id-type="doi">10.1016/j.cub.2016.10.061</article-id>
<article-categories>
<subj-group subj-group-type="heading">
<subject>Report</subject>
</subj-group>
</article-categories>
<title-group>
<article-title>17
<sup>th</sup>
Century Variola Virus Reveals the Recent History of Smallpox</article-title>
</title-group>
<contrib-group>
<contrib contrib-type="author">
<name>
<surname>Duggan</surname>
<given-names>Ana T.</given-names>
</name>
<email>duggana@mcmaster.ca</email>
<xref rid="aff1" ref-type="aff">1</xref>
<xref rid="fn1" ref-type="fn">19</xref>
<xref rid="cor1" ref-type="corresp"></xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Perdomo</surname>
<given-names>Maria F.</given-names>
</name>
<xref rid="aff2" ref-type="aff">2</xref>
<xref rid="fn1" ref-type="fn">19</xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Piombino-Mascali</surname>
<given-names>Dario</given-names>
</name>
<xref rid="aff3" ref-type="aff">3</xref>
<xref rid="fn1" ref-type="fn">19</xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Marciniak</surname>
<given-names>Stephanie</given-names>
</name>
<xref rid="aff1" ref-type="aff">1</xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Poinar</surname>
<given-names>Debi</given-names>
</name>
<xref rid="aff1" ref-type="aff">1</xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Emery</surname>
<given-names>Matthew V.</given-names>
</name>
<xref rid="aff1" ref-type="aff">1</xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Buchmann</surname>
<given-names>Jan P.</given-names>
</name>
<xref rid="aff4" ref-type="aff">4</xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Duchêne</surname>
<given-names>Sebastian</given-names>
</name>
<xref rid="aff4" ref-type="aff">4</xref>
<xref rid="aff5" ref-type="aff">5</xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Jankauskas</surname>
<given-names>Rimantas</given-names>
</name>
<xref rid="aff3" ref-type="aff">3</xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Humphreys</surname>
<given-names>Margaret</given-names>
</name>
<xref rid="aff6" ref-type="aff">6</xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Golding</surname>
<given-names>G. Brian</given-names>
</name>
<xref rid="aff7" ref-type="aff">7</xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Southon</surname>
<given-names>John</given-names>
</name>
<xref rid="aff8" ref-type="aff">8</xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Devault</surname>
<given-names>Alison</given-names>
</name>
<xref rid="aff9" ref-type="aff">9</xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Rouillard</surname>
<given-names>Jean-Marie</given-names>
</name>
<xref rid="aff9" ref-type="aff">9</xref>
<xref rid="aff10" ref-type="aff">10</xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Sahl</surname>
<given-names>Jason W.</given-names>
</name>
<xref rid="aff11" ref-type="aff">11</xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Dutour</surname>
<given-names>Olivier</given-names>
</name>
<xref rid="aff12" ref-type="aff">12</xref>
<xref rid="aff13" ref-type="aff">13</xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Hedman</surname>
<given-names>Klaus</given-names>
</name>
<xref rid="aff2" ref-type="aff">2</xref>
<xref rid="aff14" ref-type="aff">14</xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Sajantila</surname>
<given-names>Antti</given-names>
</name>
<xref rid="aff15" ref-type="aff">15</xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Smith</surname>
<given-names>Geoffrey L.</given-names>
</name>
<xref rid="aff16" ref-type="aff">16</xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Holmes</surname>
<given-names>Edward C.</given-names>
</name>
<email>edward.holmes@sydney.edu.au</email>
<xref rid="aff4" ref-type="aff">4</xref>
<xref rid="cor2" ref-type="corresp">∗∗</xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Poinar</surname>
<given-names>Hendrik N.</given-names>
</name>
<email>poinarh@mcmaster.ca</email>
<xref rid="aff1" ref-type="aff">1</xref>
<xref rid="aff7" ref-type="aff">7</xref>
<xref rid="aff17" ref-type="aff">17</xref>
<xref rid="aff18" ref-type="aff">18</xref>
<xref rid="fn2" ref-type="fn">20</xref>
<xref rid="cor3" ref-type="corresp">∗∗∗</xref>
</contrib>
</contrib-group>
<aff id="aff1">
<label>1</label>
McMaster Ancient DNA Centre, Department of Anthropology, McMaster University, Hamilton, ON L8S 4L8, Canada</aff>
<aff id="aff2">
<label>2</label>
Department of Virology, University of Helsinki, Helsinki 00014, Finland</aff>
<aff id="aff3">
<label>3</label>
Department of Anatomy, Histology, and Anthropology, Faculty of Medicine, Vilnius University, Vilnius 03101, Lithuania</aff>
<aff id="aff4">
<label>4</label>
Marie Bashir Institute for Infectious Diseases and Biosecurity, Charles Perkins Centre, School of Life and Environmental Sciences and Sydney Medical School, The University of Sydney, Sydney, NSW 2145, Australia</aff>
<aff id="aff5">
<label>5</label>
Department of Biochemistry and Molecular Biology, Bio21 Molecular Science and Biotechnology Institute, The University of Melbourne, Melbourne, VIC 3010, Australia</aff>
<aff id="aff6">
<label>6</label>
Department of History, Duke University, Durham, NC 27708-0719, USA</aff>
<aff id="aff7">
<label>7</label>
Department of Biology, McMaster University, Hamilton, ON L8S 4L8, Canada</aff>
<aff id="aff8">
<label>8</label>
Keck Carbon Cycle Accelerator Mass Spectrometer, Earth Systems Science Department, University of California, Irvine, CA 92697-3100, USA</aff>
<aff id="aff9">
<label>9</label>
MYcroarray, Ann Arbor, MI 48105, USA</aff>
<aff id="aff10">
<label>10</label>
Department of Chemical Engineering, University of Michigan, Ann Arbor, MI 48109-2136, USA</aff>
<aff id="aff11">
<label>11</label>
Center for Microbial Genetics and Genomics, Northern Arizona University, Flagstaff, AZ 86011-4073, USA</aff>
<aff id="aff12">
<label>12</label>
Laboratoire d’Anthropologie Biologique Paul Broca, Ecole Pratique des Hautes Etudes, PSL Research University, Paris 75014, France</aff>
<aff id="aff13">
<label>13</label>
PACEA, CNRS, Université de Bordeaux, Pessac 33615, France</aff>
<aff id="aff14">
<label>14</label>
Helsinki University Hospital, Helsinki 00029, Finland</aff>
<aff id="aff15">
<label>15</label>
Department of Forensic Medicine, University of Helsinki, Helsinki 00014, Finland</aff>
<aff id="aff16">
<label>16</label>
Department of Pathology, University of Cambridge, Cambridge CB2 1QP, UK</aff>
<aff id="aff17">
<label>17</label>
Michael G. DeGroote Institute for Infectious Disease Research, McMaster University, Hamilton, ON L8S 4L8, Canada</aff>
<aff id="aff18">
<label>18</label>
Humans and the Microbiome Program, Canadian Institute for Advanced Research, Toronto, ON M5G 1Z8, Canada</aff>
<author-notes>
<corresp id="cor1">
<label></label>
Corresponding author
<email>duggana@mcmaster.ca</email>
</corresp>
<corresp id="cor2">
<label>∗∗</label>
Corresponding author
<email>edward.holmes@sydney.edu.au</email>
</corresp>
<corresp id="cor3">
<label>∗∗∗</label>
Corresponding author
<email>poinarh@mcmaster.ca</email>
</corresp>
<fn id="fn1">
<label>19</label>
<p id="ntpara0010">Co-first author</p>
</fn>
<fn id="fn2">
<label>20</label>
<p id="ntpara0015">Lead Contact</p>
</fn>
</author-notes>
<pub-date pub-type="pmc-release">
<day>19</day>
<month>12</month>
<year>2016</year>
</pub-date>
<pmc-comment> PMC Release delay is 0 months and 0 days and was based on .</pmc-comment>
<pub-date pub-type="ppub">
<day>19</day>
<month>12</month>
<year>2016</year>
</pub-date>
<volume>26</volume>
<issue>24</issue>
<fpage>3407</fpage>
<lpage>3412</lpage>
<history>
<date date-type="received">
<day>26</day>
<month>9</month>
<year>2016</year>
</date>
<date date-type="rev-recd">
<day>20</day>
<month>10</month>
<year>2016</year>
</date>
<date date-type="accepted">
<day>31</day>
<month>10</month>
<year>2016</year>
</date>
</history>
<permissions>
<copyright-statement>© 2016 The Authors</copyright-statement>
<copyright-year>2016</copyright-year>
<license license-type="CC BY" xlink:href="http://creativecommons.org/licenses/by/4.0/">
<license-p>This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).</license-p>
</license>
</permissions>
<abstract id="abs0010">
<title>Summary</title>
<p>Smallpox holds a unique position in the history of medicine. It was the first disease for which a vaccine was developed and remains the only human disease eradicated by vaccination. Although there have been claims of smallpox in Egypt, India, and China dating back millennia [
<xref rid="bib1" ref-type="bibr">1</xref>
,
<xref rid="bib2" ref-type="bibr">2</xref>
,
<xref rid="bib3" ref-type="bibr">3</xref>
,
<xref rid="bib4" ref-type="bibr">4</xref>
], the timescale of emergence of the causative agent, variola virus (VARV), and how it evolved in the context of increasingly widespread immunization, have proven controversial [
<xref rid="bib4" ref-type="bibr">4</xref>
,
<xref rid="bib5" ref-type="bibr">5</xref>
,
<xref rid="bib6" ref-type="bibr">6</xref>
,
<xref rid="bib7" ref-type="bibr">7</xref>
,
<xref rid="bib8" ref-type="bibr">8</xref>
,
<xref rid="bib9" ref-type="bibr">9</xref>
]. In particular, some molecular-clock-based studies have suggested that key events in VARV evolution only occurred during the last two centuries [
<xref rid="bib4" ref-type="bibr">4</xref>
,
<xref rid="bib5" ref-type="bibr">5</xref>
,
<xref rid="bib6" ref-type="bibr">6</xref>
] and hence in apparent conflict with anecdotal historical reports, although it is difficult to distinguish smallpox from other pustular rashes by description alone. To address these issues, we captured, sequenced, and reconstructed a draft genome of an ancient strain of VARV, sampled from a Lithuanian child mummy dating between 1643 and 1665 and close to the time of several documented European epidemics [
<xref rid="bib1" ref-type="bibr">1</xref>
,
<xref rid="bib2" ref-type="bibr">2</xref>
,
<xref rid="bib10" ref-type="bibr">10</xref>
]. When compared to vaccinia virus, this archival strain contained the same pattern of gene degradation as 20
<sup>th</sup>
century VARVs, indicating that such loss of gene function had occurred before ca. 1650. Strikingly, the mummy sequence fell basal to all currently sequenced strains of VARV on phylogenetic trees. Molecular-clock analyses revealed a strong clock-like structure and that the timescale of smallpox evolution is more recent than often supposed, with the diversification of major viral lineages only occurring within the 18
<sup>th</sup>
and 19
<sup>th</sup>
centuries, concomitant with the development of modern vaccination.</p>
</abstract>
<abstract abstract-type="author-highlights" id="abs0015">
<title>Highlights</title>
<p>
<list list-type="simple">
<list-item id="u0010">
<label></label>
<p>Variola virus genome was reconstructed from a 17
<sup>th</sup>
century mummified child</p>
</list-item>
<list-item id="u0015">
<label></label>
<p>The archival strain is basal to all 20
<sup>th</sup>
century strains, with same gene degradation</p>
</list-item>
<list-item id="u0020">
<label></label>
<p>Molecular-clock analyses show that much of variola virus evolution occurred recently</p>
</list-item>
</list>
</p>
</abstract>
<abstract abstract-type="teaser" id="abs0020">
<p>Using ancient DNA sequences of variola virus recovered from the mummified remains of a 17
<sup>th</sup>
century child, Duggan et al. reconstruct the evolutionary history of smallpox. With the ancient strain, the genetic diversification of the smallpox virus is found to be more recent than previously supposed and concurrent with the onset of widespread vaccination.</p>
</abstract>
<kwd-group id="kwrds0010">
<title>Keywords</title>
<kwd>variola virus</kwd>
<kwd>smallpox</kwd>
<kwd>ancient DNA</kwd>
<kwd>Lithuanian Mummy Project</kwd>
<kwd>evolution</kwd>
<kwd>molecular clock</kwd>
<kwd>phylogeny</kwd>
</kwd-group>
</article-meta>
<notes>
<p id="misc0010">Published: December 8, 2016</p>
</notes>
</front>
<floats-group>
<fig id="fig1">
<label>Figure 1</label>
<caption>
<p>Dominican Church of the Holy Spirit, Vilnius</p>
<p>Left: Lithuania is shaded in yellow, with the red star indicating the city of Vilnius, the location of the Dominican Church of the Holy Spirit where the VD21 specimen was found and dated to approximately 1654. Dates in black indicate known smallpox outbreaks in nearby countries during the 17
<sup>th</sup>
century [
<xref rid="bib1" ref-type="bibr">1</xref>
,
<xref rid="bib2" ref-type="bibr">2</xref>
]. Top right: the Dominican Church of the Holy Spirit in Vilnius, Lithuania. Bottom right: the crypt where the child mummy was located. See also
<xref rid="mmc1" ref-type="supplementary-material">Figure S1</xref>
and
<xref rid="mmc1" ref-type="supplementary-material">Table S1</xref>
.</p>
</caption>
<graphic xlink:href="gr1"></graphic>
</fig>
<fig id="fig2">
<label>Figure 2</label>
<caption>
<p>VARV Genome Reconstructed from VD21</p>
<p>(A) Coverage of reference
<ext-link ext-link-type="uri" xlink:href="ncbi-n:NC_001611.1" id="intref0040">NC_001611.1</ext-link>
variola (major) virus genome. The inner-most circle (light purple) indicates the full 185,578 bp length of the reference, and the inner yellow circle depicts GC content across the reference genome with the genomic average of 32.7% indicated by the thin dark ring. The dark-purple ring indicates the location of annotated genes in the reference. The outer-most ring (dark orange) represents the coverage depth of reads from sample VD21 mapped to the
<ext-link ext-link-type="uri" xlink:href="ncbi-n:NC_001611.1" id="intref0045">NC_001611.1</ext-link>
reference sequence averaged across 25 bp windows. Average coverage was 18× (minimum 0× to maximum 60×). The concentric gray lines represent intervals of 10× coverage. The plot was constructed with Circos [
<xref rid="bib14" ref-type="bibr">14</xref>
].</p>
<p>(B) Conservation of genomic sequence between VD21 and the VARV reference genome
<ext-link ext-link-type="uri" xlink:href="ncbi-n:NC_001611.1" id="intref0050">NC_001611.1</ext-link>
. The plot was constructed with Dotter [
<xref rid="bib15" ref-type="bibr">15</xref>
].</p>
<p>See also
<xref rid="mmc1" ref-type="supplementary-material">Figure S2</xref>
.</p>
</caption>
<graphic xlink:href="gr2"></graphic>
</fig>
<fig id="fig3">
<label>Figure 3</label>
<caption>
<p>Evolutionary History of VARV</p>
<p>(A) Maximum-likelihood phylogeny of 43 strains of VARV rooted using homologous sequences from camels (camelpox) and African gerbils (taterapox), which are the orthopoxviruses most closely related to VARV. All horizontal branch lengths are drawn to a scale of nucleotide substitutions per site, and bootstrap values are shown for key nodes, with year of sampling shown for the VARV strains. The VD21 strain is shown in red, and the major clades of VARV (P-I and P-II, with the later the containing alastrim minor strains) are marked.</p>
<p>(B) Posterior probability densities of mean evolutionary rate estimates for VARV under different molecular-clock and coalescent models, possible
<sup>14</sup>
C dates for VD21 (i.e., “1789” assumes VD21 is from 1789 rather than 1654), and with VD21 excluded (i.e., “−VD21”) from the analysis (see
<xref rid="mmc1" ref-type="supplementary-material">Table S3</xref>
for full results).</p>
<p>(C) Maximum clade credibility tree showing the timescale of VARV evolution inferred under a strict molecular clock and a constant population size, with the VD21 lineage shown in red and the 95% credible intervals for a number of other key divergence events also shown. The date (1796) of the introduction of the cowpox-based smallpox vaccine by Jenner, which we include as an historical reference point only, is marked by a hatched line, and it is important to note that a process of smallpox variolation (inoculation) had most likely been in existence for many centuries prior to this.</p>
<p>See also
<xref rid="mmc1" ref-type="supplementary-material">Figure S3</xref>
and
<xref rid="mmc1" ref-type="supplementary-material">Tables S2</xref>
and
<xref rid="mmc1" ref-type="supplementary-material">S3</xref>
.</p>
</caption>
<graphic xlink:href="gr3"></graphic>
</fig>
</floats-group>
</pmc>
</record>

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