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

doi:10.1093/infdis/jiy629

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 journal › Article › peer-review

6 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 language	English (US)
Article number	jiy629
Pages (from-to)	187-194
Number of pages	8
Journal	Journal of Infectious Diseases
Volume	220
Issue number	2
DOIs	https://doi.org/10.1093/infdis/jiy629
State	Published - Jun 19 2019

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.

Keywords

common cold
inoculation
reverse genetics
rhinovirus

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

Publisher link

10.1093/infdis/jiy629

Find It

Find It at U of M Libraries

Cite this

Gern, J. E., Lee, W. M., Swenson, C. A., Nakagome, K., Lee, I., Wolff, M., Grindle, K., Sigelman, S., Liggett, S. B., Togias, A., Evans, M., Denlinger, L., Gangnon, R., Bochkov, Y. A., & Crisafi, G. (2019). Development of a Rhinovirus Inoculum Using a Reverse Genetics Approach. Journal of Infectious Diseases, 220(2), 187-194. Article jiy629. https://doi.org/10.1093/infdis/jiy629

@article{cf18bbce62d24b01b168ac338031e301,

title = "Development of a Rhinovirus Inoculum Using a Reverse Genetics Approach",

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.",

keywords = "common cold, inoculation, reverse genetics, rhinovirus",

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

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: {\textcopyright} 2018 The Author(s) 2018.",

year = "2019",

month = jun,

day = "19",

doi = "10.1093/infdis/jiy629",

language = "English (US)",

volume = "220",

pages = "187--194",

journal = "Journal of Infectious Diseases",

issn = "0022-1899",

publisher = "Oxford University Press",

number = "2",

}

TY - JOUR

T1 - Development of a Rhinovirus Inoculum Using a Reverse Genetics Approach

AU - Gern, James E.

AU - Lee, Wai Ming

AU - Swenson, Cheri A.

AU - Nakagome, Kazuyuki

AU - Lee, Iris

AU - Wolff, Michele

AU - Grindle, Kristine

AU - Sigelman, Steven

AU - Liggett, Stephen B.

AU - Togias, Alkis

AU - Evans, Michael

AU - Denlinger, Loren

AU - Gangnon, Ronald

AU - Bochkov, Yury A.

AU - Crisafi, Gina

N1 - 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.

PY - 2019/6/19

Y1 - 2019/6/19

N2 - 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.

AB - 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.

KW - common cold

KW - inoculation

KW - reverse genetics

KW - rhinovirus

UR - http://www.scopus.com/inward/record.url?scp=85068484320&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85068484320&partnerID=8YFLogxK

U2 - 10.1093/infdis/jiy629

DO - 10.1093/infdis/jiy629

M3 - Article

C2 - 30383246

AN - SCOPUS:85068484320

SN - 0022-1899

VL - 220

SP - 187

EP - 194

JO - Journal of Infectious Diseases

JF - Journal of Infectious Diseases

IS - 2

M1 - jiy629

ER -

Development of a Rhinovirus Inoculum Using a Reverse Genetics Approach

Abstract

Bibliographical note

Keywords

UN SDGs

Publisher link

Find It

Other files and links

Fingerprint

Cite this