Development of a Rhinovirus Inoculum Using a Reverse Genetics Approach

James E. Gern, Wai Ming Lee, Cheri A. Swenson, Kazuyuki Nakagome, Iris Lee, Michele Wolff, Kristine Grindle, Steven Sigelman, Stephen B. Liggett, Alkis Togias, Michael Evans, Loren Denlinger, Ronald Gangnon, Yury A. Bochkov, Gina Crisafi

Research output: Contribution to journalArticlepeer-review

2 Scopus citations

Abstract

Background: Experimental inoculation is an important tool for common cold and asthma research. Producing rhinovirus (RV) inocula from nasal secretions has required prolonged observation of the virus donor to exclude extraneous pathogens. We produced a RV-A16 inoculum using reverse genetics and determined the dose necessary to cause moderate colds in seronegative volunteers. Methods: The consensus sequence of RV-A16 from a previous inoculum was cloned, and inoculum virus was produced using reverse genetics techniques. After safety testing, volunteers were inoculated with either RV-A16 (n = 26) or placebo (n = 10), Jackson cold scores were recorded, and nasal secretions were tested for shedding of RV-A16 ribonucleic acid. Results: The reverse genetics process produced infectious virus that was neutralized by specific antisera and had a mutation rate similar to conventional virus growth techniques. The 1000 median tissue culture infectious dose (TCID50) dose produced moderate colds in most individuals with effects similar to that of a previously tested conventional RV-A16 inoculum. Conclusions: Reverse genetics techniques produced a RV-A16 inoculum that can cause clinical colds in seronegative volunteers, and they also serve as a stable source of virus for laboratory use. The recombinant production procedures eliminate the need to derive seed virus from nasal secretions, thus precluding introduction of extraneous pathogens through this route.

Original languageEnglish (US)
Article numberjiy629
Pages (from-to)187-194
Number of pages8
JournalJournal of Infectious Diseases
Volume220
Issue number2
DOIs
StatePublished - Jun 19 2019
Externally publishedYes

Bibliographical note

Funding Information:
Potential conflicts of interest. J. E. G. reports the following: grants from NIH during the conduct of the study; and personal fees from PREP Biopharm Inc., Regeneron, MedImmune, and Meissa Vaccines Inc. outside the submitted work. L. D. reports grants from NIH during the conduct of the study and personal fees from AstraZeneca and Sanofi outside the submitted work. R. G., M. E., and Y. A. B. report grants from NIH during the conduct of the study. J. E. G. is a consultant to Regeneron, PREP Biopharma, and Meissa Vaccines. L. D. is a consultant to AstraZeneca and Sanofi. All authors have submitted the ICMJE Form for Disclosure of Potential Conflicts of Interest. Conflicts that the editors consider relevant to the content of the manuscript have been disclosed.

Funding Information:
Acknowledgments. We thank John Centanni (Institute for Clinical and Translational Research, University of Wisconsin-Madison) for assistance in preparing The Investigational New Drug application to the US Food and Drug Administration. Fincancial support. This work was funded by the National Institute of Allergy and Infectious Diseases through Grant Numbers U19 AI070503 and U19 AI104317 and the Clinical and Translational Science Award program through the National Institutes of Health (NIH), National Center for Advancing Translational Sciences, Grant UL1TR000427.

Publisher Copyright:
© 2018 The Author(s) 2018.

Copyright:
Copyright 2021 Elsevier B.V., All rights reserved.

Keywords

  • common cold
  • inoculation
  • reverse genetics
  • rhinovirus

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