Infectious virus in exhaled breath of symptomatic seasonal influenza cases from a college community

Jing Yan; Michael Grantham; Jovan Pantelic; P. Jacob Bueno de Mesquita; Barbara Albert; Fengjie Liu; Sheryl Ehrman; Donald K. Milton; EMIT Consortium

doi:10.1073/pnas.1716561115

New Research In

Edited by Peter Palese, Icahn School of Medicine at Mount Sinai, New York, NY, and approved December 15, 2017 (received for review September 19, 2017)

Significance

Lack of human data on influenza virus aerosol shedding fuels debate over the importance of airborne transmission. We provide overwhelming evidence that humans generate infectious aerosols and quantitative data to improve mathematical models of transmission and public health interventions. We show that sneezing is rare and not important for—and that coughing is not required for—influenza virus aerosolization. Our findings, that upper and lower airway infection are independent and that fine-particle exhaled aerosols reflect infection in the lung, opened a pathway for a deeper understanding of the human biology of influenza infection and transmission. Our observation of an association between repeated vaccination and increased viral aerosol generation demonstrated the power of our method, but needs confirmation.

Abstract

Little is known about the amount and infectiousness of influenza virus shed into exhaled breath. This contributes to uncertainty about the importance of airborne influenza transmission. We screened 355 symptomatic volunteers with acute respiratory illness and report 142 cases with confirmed influenza infection who provided 218 paired nasopharyngeal (NP) and 30-minute breath samples (coarse >5-µm and fine ≤5-µm fractions) on days 1–3 after symptom onset. We assessed viral RNA copy number for all samples and cultured NP swabs and fine aerosols. We recovered infectious virus from 52 (39%) of the fine aerosols and 150 (89%) of the NP swabs with valid cultures. The geometric mean RNA copy numbers were 3.8 × 10⁴/30-minutes fine-, 1.2 × 10⁴/30-minutes coarse-aerosol sample, and 8.2 × 10⁸ per NP swab. Fine- and coarse-aerosol viral RNA were positively associated with body mass index and number of coughs and negatively associated with increasing days since symptom onset in adjusted models. Fine-aerosol viral RNA was also positively associated with having influenza vaccination for both the current and prior season. NP swab viral RNA was positively associated with upper respiratory symptoms and negatively associated with age but was not significantly associated with fine- or coarse-aerosol viral RNA or their predictors. Sneezing was rare, and sneezing and coughing were not necessary for infectious aerosol generation. Our observations suggest that influenza infection in the upper and lower airways are compartmentalized and independent.

Footnotes

↵¹Present address: Department of Biology, Missouri Western State University, St. Joseph, MO 64507.
↵²Present address: Center for the Built Environment, University of California, Berkeley, CA 94720.
↵³Present address: Division of Viral Products, Center for Biologics Evaluation and Research, Food and Drug Administration, Silver Spring, MD 20903.
↵⁴Present address: Davidson College of Engineering, San José State University, San José, CA 95192.
↵⁵To whom correspondence should be addressed. Email: dmilton{at}umd.edu.
↵⁶A complete list of the EMIT Consortium can be found in the Supporting Information.

Author contributions: S.E., D.K.M., and E.C. designed research; J.Y., M.G., J.P., P.J.B.d.M., B.A., F.L., S.E., and D.K.M. performed research; D.K.M. contributed new reagents/analytic tools; J.Y. and D.K.M. analyzed data; and J.Y., M.G., and D.K.M. wrote the paper.
The authors declare no conflict of interest.
This article is a PNAS Direct Submission.
This article contains supporting information online at www.pnas.org/lookup/suppl/doi:10.1073/pnas.1716561115/-/DCSupplemental.

This open access article is distributed under Creative Commons Attribution-NonCommercial-NoDerivatives License 4.0 (CC BY-NC-ND).

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