Single-Cell Analysis of Nasal Mucosa Reveals Immune Response in Respiratory Syncytial Virus (RSV) Infection

Authors

Santosh Khanal, Children's Mercy HospitalFollow
Eric S. Geanes, Children's Mercy HospitalFollow
Carl F. Schreck, Children's Mercy Kansas CityFollow
Daniel A. Louiselle, Children's Mercy HospitalFollow
Alyse Peters, Children's Mercy Kansas CityFollow
Nick Nolte, Children's Mercy Kansas CityFollow
Libby Corkran, Children's Mercy Kansas CityFollow
Anjana Sasidharan, Children's Mercy HospitalFollow
Bradley Belden, Children's Mercy HospitalFollow
Gage Greening, Children's Mercy Kansas CityFollow
Rebecca McLennan, Children's Mercy HospitalFollow
Tomi Pastinen, Children's Mercy Kansas CityFollow
Rangaraj Selvarangan, Children's Mercy HospitalFollow
Todd Bradley, Children's Mercy HospitalFollow
Elin Grundberg, Children's Mercy HospitalFollow

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Publication Date

5-2025

Abstract

Respiratory Syncytial Virus (RSV) is a common respiratory pathogen that can cause breathing problems in babies and young kids, sometimes leading to serious health issues. Insight into pathologic immune responses and cellular dysregulations that contribute to disease severity has largely relied on invasive sampling typically restricted to critically ill and mechanically ventilated children. However, the nasal mucosa is a critical site for viral entry, making it an ideal target for studying viral pathogenesis and allows sample collection noninvasively. Here, we present an approach for simultaneous single-cell profiling and viral typing using salvaged nasal swabs obtained in pediatric clinics. First, we developed a comprehensive viral and human reference genome panel applied to 80,970 nasal cells, and mapped not only RSV, but also influenza A and B and SARS-CoV-2, infected and non-infected epithelial and immune cells. Compared to other viruses, we identified macrophages and neutrophils as the primary cellular target of RSV in the nasal mucosa and characterized a significant increase in IL-8 expressing neutrophils during infection. This was accompanied by a significant decline in ciliated cells in RSV samples compared to controls. Consistent with these findings, pathway analysis revealed significant activation of immune-related, particularly TNFα signaling via NFκB and interferon gamma response, and complement cascade pathways in both cell types, while metabolic processes like glycolysis and protein secretion showed less significant changes.

In conclusion, our study provides insights into proinflammatory roles of macrophages and neutrophils in the nasal mucosa after RSV infection. These findings underscore the utility of single-cell genomics in unraveling complex host-pathogen interactions, highlighting potential for personalized medicine in pediatric respiratory diseases. Follow up experiments focusing on confirming our single cell sequencing findings using histological approaches and protein analyses are currently ongoing.

Document Type

Poster

Recommended Citation

Khanal, Santosh; Geanes, Eric S.; Schreck, Carl F.; Louiselle, Daniel A.; Peters, Alyse; Nolte, Nick; Corkran, Libby; Sasidharan, Anjana; Belden, Bradley; Greening, Gage; McLennan, Rebecca; Pastinen, Tomi; Selvarangan, Rangaraj; Bradley, Todd; and Grundberg, Elin, "Single-Cell Analysis of Nasal Mucosa Reveals Immune Response in Respiratory Syncytial Virus (RSV) Infection" (2025). Research at Children's Mercy Month 2025. 1.
https://scholarlyexchange.childrensmercy.org/research_month2025/1