Deliberate Attenuation of Chikungunya Virus by Adaptation to Heparan Sulfate-Dependent Infectivity

A Model for Rational Arboviral Vaccine Design

Christina L. Gardner, Jozef Hritz, Chengqun Sun, Dana L. Vanlandingham, Timothy Y. Song, Elodie Ghedin, Stephen Higgs, William B. Klimstra, Kate D. Ryman

Research output: Contribution to journalArticle

Abstract

Mosquito-borne chikungunya virus (CHIKV) is a positive-sense, single-stranded RNA virus from the genus Alphavirus, family Togaviridae, which causes fever, rash and severe persistent polyarthralgia in humans. Since there are currently no FDA licensed vaccines or antiviral therapies for CHIKV, the development of vaccine candidates is of critical importance. Historically, live-attenuated vaccines (LAVs) for protection against arthropod-borne viruses have been created by blind cell culture passage leading to attenuation of disease, while maintaining immunogenicity. Attenuation may occur via multiple mechanisms. However, all examined arbovirus LAVs have in common the acquisition of positively charged amino acid substitutions in cell-surface attachment proteins that render virus infection partially dependent upon heparan sulfate (HS), a ubiquitously expressed sulfated polysaccharide, and appear to attenuate by retarding dissemination of virus particles in vivo. We previously reported that, like other wild-type Old World alphaviruses, CHIKV strain, La Réunion, (CHIKV-LR), does not depend upon HS for infectivity. To deliberately identify CHIKV attachment protein mutations that could be combined with other attenuating processes in a LAV candidate, we passaged CHIKV-LR on evolutionarily divergent cell-types. A panel of single amino acid substitutions was identified in the E2 glycoprotein of passaged virus populations that were predicted to increase electrostatic potential. Each of these substitutions was made in the CHIKV-LR cDNA clone and comparisons of the mutant viruses revealed surface exposure of the mutated residue on the spike and sensitivity to competition with the HS analog, heparin, to be primary correlates of attenuation in vivo. Furthermore, we have identified a mutation at E2 position 79 as a promising candidate for inclusion in a CHIKV LAV.

Original languageEnglish (US)
Article numbere2719
JournalPLoS Neglected Tropical Diseases
Volume8
Issue number2
DOIs
StatePublished - 2014

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Chikungunya virus
Heparitin Sulfate
Vaccines
Attenuated Vaccines
Alphavirus
Arboviruses
Amino Acid Substitution
Togaviridae
Viruses
Virus Attachment
Mutation
RNA Viruses
Arthralgia
Virus Diseases
Exanthema
Culicidae
Static Electricity
Virion
Antiviral Agents
Polysaccharides

ASJC Scopus subject areas

  • Infectious Diseases
  • Public Health, Environmental and Occupational Health
  • Pharmacology, Toxicology and Pharmaceutics(all)

Cite this

Deliberate Attenuation of Chikungunya Virus by Adaptation to Heparan Sulfate-Dependent Infectivity : A Model for Rational Arboviral Vaccine Design. / Gardner, Christina L.; Hritz, Jozef; Sun, Chengqun; Vanlandingham, Dana L.; Song, Timothy Y.; Ghedin, Elodie; Higgs, Stephen; Klimstra, William B.; Ryman, Kate D.

In: PLoS Neglected Tropical Diseases, Vol. 8, No. 2, e2719, 2014.

Research output: Contribution to journalArticle

Gardner, Christina L. ; Hritz, Jozef ; Sun, Chengqun ; Vanlandingham, Dana L. ; Song, Timothy Y. ; Ghedin, Elodie ; Higgs, Stephen ; Klimstra, William B. ; Ryman, Kate D. / Deliberate Attenuation of Chikungunya Virus by Adaptation to Heparan Sulfate-Dependent Infectivity : A Model for Rational Arboviral Vaccine Design. In: PLoS Neglected Tropical Diseases. 2014 ; Vol. 8, No. 2.
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abstract = "Mosquito-borne chikungunya virus (CHIKV) is a positive-sense, single-stranded RNA virus from the genus Alphavirus, family Togaviridae, which causes fever, rash and severe persistent polyarthralgia in humans. Since there are currently no FDA licensed vaccines or antiviral therapies for CHIKV, the development of vaccine candidates is of critical importance. Historically, live-attenuated vaccines (LAVs) for protection against arthropod-borne viruses have been created by blind cell culture passage leading to attenuation of disease, while maintaining immunogenicity. Attenuation may occur via multiple mechanisms. However, all examined arbovirus LAVs have in common the acquisition of positively charged amino acid substitutions in cell-surface attachment proteins that render virus infection partially dependent upon heparan sulfate (HS), a ubiquitously expressed sulfated polysaccharide, and appear to attenuate by retarding dissemination of virus particles in vivo. We previously reported that, like other wild-type Old World alphaviruses, CHIKV strain, La R{\'e}union, (CHIKV-LR), does not depend upon HS for infectivity. To deliberately identify CHIKV attachment protein mutations that could be combined with other attenuating processes in a LAV candidate, we passaged CHIKV-LR on evolutionarily divergent cell-types. A panel of single amino acid substitutions was identified in the E2 glycoprotein of passaged virus populations that were predicted to increase electrostatic potential. Each of these substitutions was made in the CHIKV-LR cDNA clone and comparisons of the mutant viruses revealed surface exposure of the mutated residue on the spike and sensitivity to competition with the HS analog, heparin, to be primary correlates of attenuation in vivo. Furthermore, we have identified a mutation at E2 position 79 as a promising candidate for inclusion in a CHIKV LAV.",
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