Evolution of influenza genome diversity during infection in immunocompetent patients

Abstract

IntroductionMinor frequency viruses play many important roles during viral infection that cannot be explained by the consensus sequence alone. In influenza, immunosuppressed individuals appear to generate numerous viral variants, leading to subpopulations with important role in infection. The objective of the present study was to describe viral diversification over time in immunocompetent patients during influenza virus infection.\n\nMethodsAll clinical records of patients admitted to the Lyon university hospital (Lyon, France) during the influenza infection epidemics of the 2010-2015 period and sampled at least twice during their clinical management were retrospectively analyzed. To estimate performance of the sequencing procedures, well-characterized plasmids containing each of the 8 segments of influenza viruses were used as quality controls. Diversity, i.e. the number of validated single nucleotide variants, was analyzed to compare characteristics over time and according to clinical severity (mild, severe with neurological complications, severe with respiratory complications).\n\nResultsAfter validation on quality controls (n=51), and verification of possible confusion bias, a 5%-threshold of detection was applied to clinical viral sequences (n=29). At this threshold, amino-acid coordinates (n=183/4,246, 4.31%) were identified as having at least one mutation during clinical course, independently of the clinical severity. Considering a threshold of 4 days of symptoms, as a limit for early and late sampling, diversity was significantly higher in late samples for the mild group, compared to both early mild and severe groups (p<0.05). At a single-segment scale, for PB2-coding segment, diversity was significantly higher in early samples of the neurological group than in both early and late samples in the respiratory group and for late samples in the mild group (p<0.05). For the NS1-coding segment, significant differences were observed between initial diversity of mild and severe patients, as for early and late samples in mild patients (p<0.01). Discussion. This study is the first describing diversity through time, associating biological and clinical information during viral diversification, during the infection of an immunocompetent human host. This latter opens a large field of investigation in infectious disease management using next-generation sequencing and suggest development of new therapies, focusing on non-antigenic viral properties, in non-vaccine fields of research