Abstract
Human respiratory syncytial virus (HRSV) is a major cause of pediatric infection and also causes disease in the elderly and those with underlying respiratory problems. There is no vaccine for HRSV and anti-viral therapeutics are not broadly applicable. To investigate the effect of HRSV biology in children, nasopharyngeal aspirates were taken from children with different viral loads and a combined high throughput RNAseq and label free quantitative proteomics approach was used to characterize the nucleic acid and proteins in these samples. HRSV proteins were identified in the nasopharyngeal aspirates from infected children, and their abundance correlated with viral load (Ct value), confirming HRSV infection. Analysis of the HRSV genome indicated that the children were infected with sub-group A virus and that minor variants in nucleotide frequency occurred in discrete clusters along the HRSV genome, and within a patient clustered distinctly within the glycoprotein gene. Data from the samples were binned into four groups; no-HRSV infection (control), high viral load (Ct < 20), medium viral load (Ct = 20–25), and low viral load (Ct > 25). Cellular proteins associated with the anti-viral response (e.g., ISG15) were identified in the nasopharyngeal aspirates and their abundance was correlated with viral load. These combined approaches have not been used before to study HRSV biology in vivo and can be readily applied to the study the variation of virus host interactions.
Highlights
Human respiratory syncytial virus (HRSV) is one of the major lower respiratory tract pathogens in infants with consistent annual outbreaks. 90% of infants are infected with HRSV at least once within the first two years of life [1,2]
To investigate the effects of HRSV infection in this patient group, nasopharyngeal aspirates that had been taken for routine diagnostic purposes, were analyzed using quantitative proteomics and RNA sequencing (RNAseq)
For the children infected with HRSV, samples were analyzed from children with different viral loads at the time of sampling, and these were grouped into three arbitrary categories; low viral load (Ct > 25) (n = 7), medium viral load (Ct = 20–25) (n = 3), and high viral load (Ct < 20) (n = 2)
Summary
Human respiratory syncytial virus (HRSV) is one of the major lower respiratory tract pathogens in infants with consistent annual outbreaks. 90% of infants are infected with HRSV at least once within the first two years of life [1,2]. 90% of infants are infected with HRSV at least once within the first two years of life [1,2]. Viruses 2019, 11, 926 and B, which represent two lines of divergent evolution with both genetic and serologic differences. While both subgroup A and B viruses contribute to overall HRSV disease, some studies have suggested that subgroup A isolates may have slightly different clinical presentations [9] and effects on host cell signaling [10] compared to subgroup B viruses. Other studies have found no difference in the severity of illness caused by either subgroup [11]. A subgroup A clade virus was shown to be associated with more severe illness than other subgroup A clades [11]
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