title: Dual RNA-seq reveals viral infections in asthmatic children without respiratory illness which are associated with changes in the airway transcriptome.
abstract: Respiratory illness caused by viral infection is associated with the development and exacerbation of childhood asthma. Little is known about the effects of respiratory viral infections in the absence of illness. Using quantitative PCR (qPCR) for common respiratory viruses and for two genes known to be highly upregulated in viral infections (CCL8/CXCL11), we screened 92 asthmatic and 69 healthy children without illness for respiratory virus infections.
  
  
  
  We found 21 viral qPCR-positive and 2 suspected virus-infected subjects with high expression of CCL8/CXCL11. We applied a dual RNA-seq workflow to these subjects, together with 25 viral qPCR-negative subjects, to compare qPCR with sequencing-based virus detection and to generate the airway transcriptome for analysis. RNA-seq virus detection achieved 86% sensitivity when compared to qPCR-based screening. We detected additional respiratory viruses in the two CCL8/CXCL11-high subjects and in two of the qPCR-negative subjects. Viral read counts varied widely and were used to stratify subjects into Virus-High and Virus-Low groups. Examination of the host airway transcriptome found that the Virus-High group was characterized by immune cell airway infiltration, downregulation of cilia genes, and dampening of type 2 inflammation. Even the Virus-Low group was differentiated from the No-Virus group by 100 genes, some involved in eIF2 signaling.
  
  
  
  Respiratory virus infection without illness is not innocuous but may determine the airway function of these subjects by driving immune cell airway infiltration, cellular remodeling, and alteration of asthmogenic gene expression.
authors.affiliation: Center for Genes, Environment, and Health, National Jewish Health, Denver, CO, USA.
authors.forename: Agata
authors.initials: A
authors.lastname: Wesolowska-Andersen
authors.affiliation: Center for Genes, Environment, and Health, National Jewish Health, Denver, CO, USA.
authors.forename: Jamie L
authors.initials: JL
authors.lastname: Everman
authors.affiliation: Center for Genes, Environment, and Health, National Jewish Health, Denver, CO, USA.
authors.forename: Rebecca
authors.initials: R
authors.lastname: Davidson
authors.affiliation: Center for Genes, Environment, and Health, National Jewish Health, Denver, CO, USA.
authors.forename: Cydney
authors.initials: C
authors.lastname: Rios
authors.affiliation: Center for Genes, Environment, and Health, National Jewish Health, Denver, CO, USA.
authors.forename: Rachelle
authors.initials: R
authors.lastname: Herrin
authors.affiliation: Department of Medicine, University of California, San Francisco, CA, USA.
authors.forename: Celeste
authors.initials: C
authors.lastname: Eng
authors.affiliation: Department of Medicine, National Jewish Health, Denver, CO, USA.
authors.forename: William J
authors.initials: WJ
authors.lastname: Janssen
authors.affiliation: Department of Pediatrics, National Jewish Health, 1400 Jackson St, Denver, CO, 80206, USA.|Children's Hospital Colorado and University of Colorado School of Medicine, Aurora, CO, USA.
authors.forename: Andrew H
authors.initials: AH
authors.lastname: Liu
authors.affiliation: Department of Medicine, University of California, San Francisco, CA, USA.
authors.forename: Sam S
authors.initials: SS
authors.lastname: Oh
authors.affiliation: Department of Pediatrics, The Ann and Robert H. Lurie Children's Hospital of Chicago, Northwestern University Feinberg School of Medicine, Chicago, IL, USA.
authors.forename: Rajesh
authors.initials: R
authors.lastname: Kumar
authors.affiliation: Center for Genes, Environment, and Health, National Jewish Health, Denver, CO, USA.|Department of Biomedical Research, National Jewish Health, Denver, CO, USA.
authors.forename: Tasha E
authors.initials: TE
authors.lastname: Fingerlin
authors.affiliation: Centro de Neumologia Pediatrica, San Juan, Puerto Rico.
authors.forename: Jose
authors.initials: J
authors.lastname: Rodriguez-Santana
authors.affiliation: Department of Medicine, University of California, San Francisco, CA, USA.|Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, CA, USA.
authors.forename: Esteban G
authors.initials: EG
authors.lastname: Burchard
authors.affiliation: Center for Genes, Environment, and Health, National Jewish Health, Denver, CO, USA. seiboldm@njhealth.org.|Department of Pediatrics, National Jewish Health, 1400 Jackson St, Denver, CO, 80206, USA. seiboldm@njhealth.org.|Division of Pulmonary Sciences and Critical Care Medicine, University of Colorado School of Medicine, Aurora, CO, USA. seiboldm@njhealth.org.
authors.forename: Max A
authors.initials: MA
authors.lastname: Seibold
mesh_terms: D001249:Asthma / Q000150:complications / Q000235:genetics* / Q000276:immunology; D016022:Case-Control Studies; D002648:Child; D020869:Gene Expression Profiling; D015967:Gene Expression Regulation, Viral; D059014:High-Throughput Nucleotide Sequencing; D054884:Host-Pathogen Interactions / Q000235:genetics* / Q000276:immunology; D006801:Humans; D012327:RNA Virus Infections / Q000150:complications / Q000235:genetics* / Q000276:immunology / Q000821:virology*; D012328:RNA Viruses / Q000235:genetics; D020545:Respiratory Mucosa / Q000276:immunology / Q000378:metabolism / Q000821:virology; D012141:Respiratory Tract Infections / Q000150:complications / Q000235:genetics* / Q000276:immunology / Q000821:virology*; D017423:Sequence Analysis, RNA; D059467:Transcriptome
keywords: Airway epithelium; Asthma; Asymptomatic; Children; Host response; Host-virus interactions; Infection; RNA-seq; Transcriptome; Virus