Using Zebrafish Models of Human Influenza A Virus Infections to Screen Antiviral Drugs and Characterize Host Immune Cell Responses

Con Sullivan; Denise Jurcyzszak; Michelle F. Goody; Kristin A. Gabor; Jacob R. Longfellow; Paul J. Millard; Carol H. Kim

doi:10.3791/55235

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Method Article

Using Zebrafish Models of Human Influenza A Virus Infections to Screen Antiviral Drugs and Characterize Host Immune Cell Responses

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DOI:

10.3791/55235

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January 20th, 2017

Con Sullivan¹^,² , Denise Jurcyzszak¹ , Michelle F. Goody³ , Kristin A. Gabor⁴ , Jacob R. Longfellow¹ , Paul J. Millard²^,⁵ , Carol H. Kim¹^,²

¹Department of Molecular and Biomedical Sciences, University of Maine, ²Graduate School of Biomedical Sciences and Engineering, University of Maine, ³School of Biology and Ecology, University of Maine, ⁴Division of Intramural Research, Immunity, Inflammation and Disease Laboratory, National Institute of Environmental Health Sciences, NIH, ⁵Department of Chemical and Biological Engineering, University of Maine

Summary

Systemic and localized zebrafish infection models for human influenza A virus are demonstrated. Using a systemic infection model, zebrafish can be used to screen antiviral drugs. Using a localized infection model, zebrafish can be used to characterize host immune cell responses.

Abstract

Each year, seasonal influenza outbreaks profoundly affect societies worldwide. In spite of global efforts, influenza remains an intractable healthcare burden. The principle strategy to curtail infections is yearly vaccination. In individuals who have contracted influenza, antiviral drugs can mitigate symptoms. There is a clear and unmet need to develop alternative strategies to combat influenza. Several animal models have been created to model host-influenza interactions. Here, protocols for generating zebrafish models for systemic and localized human influenza A virus (IAV) infection are described. Using a systemic IAV infection model, small molecules with potential antiviral activity can be screened. As a proof-of-principle, a protocol that demonstrates the efficacy of the antiviral drug Zanamivir in IAV-infected zebrafish is described. It shows how disease phenotypes can be quantified to score the relative efficacy of potential antivirals in IAV-infected zebrafish. In recent years, there has been increased appreciation for the critical role neutrophils play in the human host response to influenza infection. The zebrafish has proven to be an indispensable model for the study of neutrophil biology, with direct impacts on human medicine. A protocol to generate a localized IAV infection in the Tg(mpx:mCherry) zebrafish line to study neutrophil biology in the context of a localized viral infection is described. Neutrophil recruitment to localized infection sites provides an additional quantifiable phenotype for assessing experimental manipulations that may have therapeutic applications. Both zebrafish protocols described faithfully recapitulate aspects of human IAV infection. The zebrafish model possesses numerous inherent advantages, including high fecundity, optical clarity, amenability to drug screening, and availability of transgenic lines, including those in which immune cells such as neutrophils are labeled with fluorescent proteins. The protocols detailed here exploit these advantages and have the potential to reveal critical insights into host-IAV interactions that may ultimately translate into the clinic.

Introduction

According to the World Health Organization (WHO), influenza viruses infect 5-10% of adults and 20-30% of children annually and cause 3-5 million cases of severe illness and up to 500,000 deaths worldwide¹. Yearly vaccinations against influenza remain the best option to prevent disease. Efforts like the WHO Global Action Plan have increased seasonal vaccine use, vaccine production capacity, and research and development into more potent vaccine strategies in order to reduce morbidity and mortality associated with seasonal influenza outbreaks². Antiviral drugs like neuraminidase inhibitors (e.g. Zanamivir and Oselt....