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Koelle, K.

Publications and source records attributed to Koelle, K..

At least 19 recordsLinked to original sources

Viral dispersal drives changes in population composition

In an acute viral infection, a small seeding population expands rapidly over several orders of magnitude, then declines. This vast population growth is accompanied by spatial expansion as infection spreads to new anatomical sites within the host. Evolutionary theory predicts that spatially expanding populations experience more extreme stochastic losses of genetic diversity than spatially unstructured populations. To test this prediction in the context of viral dynamics, we evaluated the impact of spatial expansion on viral population genetic structure and assessed how modulators of viral dispersal affected this structure. To this end, we developed a means of implementing the spatial expansion of influenza A virus in cell culture and applied it to a viral population carrying a diverse genetic barcode. Across this simplified cellular landscape, we found that spatial expansion produces a patchwork of distinct viral subpopulations. We then evaluated the consequences for population structure of two modulators of viral dispersal: extracellular viscosity and a common polymorphism in the viral attachment protein. Both physical constraint and viral phenotype altered the extent of genetic bottlenecking and spatial heterogeneity that developed during expansion, indicating that environmental features and viral traits that modulate dispersal also modulate viral population composition, diversity and evolution.

microbiology↗

Evolutionary constraints limit additional mammalian adaptation of bovine-derived H5N1 influenza viruses in ferrets

The outbreak of clade 2.3.4.4b H5N1 viruses among U.S. dairy cattle has raised concerns that sustained circulation among agricultural mammals could facilitate viral adaptation toward efficient human transmission. However, the evolutionary dynamics governing such adaptation remain poorly understood. Here we investigated the evolution of two bovine-derived H5N1 B3.13 genotype viruses during infection and airborne transmission in ferrets, building on prior characterization of their robust replication and inefficient airborne transmission. Within hosts, viral genetic diversity was limited and viruses were subject to genetic drift and weak purifying selection. Transmission, when it occurred, was characterized by stringent bottlenecks that sharply reduced viral genetic diversity. We found no evidence of mammalian adaptation during infection or transmission. Together, these findings indicate that bovine-derived H5N1 viruses face evolutionary constraints during acute mammalian infection and transmission, limiting movement toward enhanced airborne spread. These constraints may help explain why efficient mammalian replication does not necessarily coincide with efficient transmission. Continued circulation of HA clade 2.3.4.4b viruses nevertheless creates repeated opportunities for rare but consequential evolutionary events, underscoring the importance of sustained surveillance and risk mitigation.

microbiology↗

Environmental stochasticity can account for patterns of within-host respiratory virus evolution

The ecological and evolutionary dynamics of populations, including viral populations, are known to be jointly shaped by deterministic and stochastic processes. While the impact of stochastic processes has been rigorously explored for viral dynamics at the level of the host population, most dynamic models for acutely-infecting respiratory viral pathogens at the within-host scale remain deterministic in their formulation. While this may be reasonable for identifying key processes shaping their within-host viral population dynamics, recent studies indicate that stochastic processes need to be invoked for understanding patterns of within-host viral evolution. Specifically, several studies have shown that viral allele frequencies can change dramatically over the time course of days in acute infections. Here, we use stochastic dynamic models to explore the role of environmental noise in shaping observed patterns of virus evolution in acute respiratory virus infections. We summarize ways in which environmental stochasticity can be biologically realized in these acute viral infections and describe within-host models that can be implemented to jointly yield viral population dynamics and evolutionary dynamics. We further develop a statistical approach to estimate the extent of environmental noise from observed within-host allele frequency changes. We test this approach on simulated data and apply it to existing influenza A virus and SARS-CoV-2 within-host data. With these applications, we show that environmental stochasticity can parsimoniously reproduce key features of empirically observed allele frequency changes without needing to invoke demographic stochasticity or to adopt Wright-Fisher model formulations with a constant effective population size. Finally, we show that purifying selection and positive selection can both still contribute to within-host viral evolution in the context of a noisy environment, providing theoretical support for studies that have found purifying and positive selection in acutely-infecting respiratory virus populations.

evolutionary biology↗

Within-host adaptive evolution is limited by genetic drift in experimental human influenza A virus infections

Selection of advantageous mutations drives the emergence of dominant variants during seasonal influenza epidemics. However, within-host detection of such variants remains rare, limiting our understanding of how selection operates at the scale of individual hosts. In this study, we used a controlled human infection model to examine the within-host evolutionary dynamics in thirteen participants intranasally infected with a seasonal H3N2 influenza A virus. Although this clinical trial is ongoing, our work represents a pre-planned, interim, exploratory analysis. Results in this system were contrasted with those observed in a ferret model of infection. The inoculum, used in both humans and ferrets, carried standing diversity that enabled evaluation of variant trajectories during infection. Although the dynamics were variable among participants, in humans, the minor variants in the PA and NP gene segments tended to increase in frequency as infection progressed. Variant dynamics were more consistent among ferrets but showed differences from humans in the fate of the minor NP allele. Based on these observations, we fit a population genetic model to longitudinal measurements of variant frequencies. Estimates of variant selection coefficients and effective viral population sizes indicated that in humans the two minor variants had a selective advantage over the major variants, but genetic drift was strong, limiting the efficiency of selection. In ferrets, the PA minor variant also was estimated to have a selective advantage, while the NP minor variant was estimated to have a selective disadvantage. Moreover, effective viral population sizes were estimated to be considerably higher in ferrets than in humans, indicating that genetic drift was weaker in ferrets. Our analyses reveal differing selective environments acting on influenza viruses in human and ferret hosts and indicate that selection at the within-host level is weakened by genetic drift.

microbiology↗

Quantifying the role of pre-existing tissue resident cellular immunity in limiting respiratory virus transmission

Viral transmission from infected donors to uninfected recipients is the key event underlying the spread of viral pathogens at the level of a host population. Successful viral transmission from a donor to a recipient depends on several factors including the infectiousness of the donor. Donor infectiousness in turn can depend on the viral kinetics and viral load of the donor, donor behavior and symptoms, and donor immunity. Here, we use a mouse model of murine respirovirus (otherwise known as Sendai virus SeV) infection to quantitatively explore donor determinants of respiratory virus transmission. The experimental transmission studies we analyze are specifically designed to address the effect that pre-existing donor immunity may have on transmission potential by studying SeV transmission from both immunized and control (placebo-immunized) donors to naive recipients. We specifically focus on the impact of tissue resident memory (TRM) CD8 T cells on donor transmission potential by considering immunization strategies that primarily generate CD8 T cell immunity. Through quantitative analyses of these experiments, we find that pre-existing CD8 TRMs act to reduce donor transmission potential. This reduction can be in part explained by a reduction in total infection load in immunized donors. However, even once differences in infection load between immunized and control donors are accounted for, immunized donors still have reduced infectiousness relative to control donors. We explore possible reasons for this unexpected pattern using a mathematical model that integrates within-host viral dynamics and between-host transmission occurrences. Analysis of model simulations, along with observations from knock-out experiments, suggests that interferon gamma (IFN-{gamma}) may be partly responsible for the observed differences in infectiousness between control and pre-immune donors. Future experimental transmission studies should consider measuring IFN-{gamma} levels when interpreting transmission outcomes in the context of host immunity.

immunology↗

Genetic drift acts strongly on within-host influenza virus populations during acute infection but does not act alone

The evolutionary dynamics of seasonal influenza A viruses (IAVs) have been well characterized at the population level, with antigenic drift known to be a major force in driving strain turnover. The evolution of IAV populations at the within-host level, however, is still less well characterized. Improving our understanding of within-host IAV evolution has the potential to shed light on the source of new strains, including new antigenic variants, at the population level. Existing studies have pointed towards the role that stochastic processes play in shaping within-host viral evolution in acute infections of both humans and pigs. Here, we apply a population genetic model called the Beta-with-Spikes approximation to longitudinal intrahost Single Nucleotide Variant (iSNV) frequency data to quantify the extent of genetic drift acting on IAV populations at the within-host scale. We estimate small effective population sizes in both human IAV infections (NE = 41, 95% confidence interval: [22-72]) and swine IAV infections (NE = 10, 95% confidence interval: [8-14]). Moreover, we evaluate the consistency of the observed iSNV dynamics with Wright-Fisher model simulations. For the human IAV dataset that we analyze, we find that observed within-host IAV evolutionary dynamics are consistent with this classic model at the estimated low effective population size. However, for the swine IAV dataset, we find statistical evidence for rejecting the classic Wright-Fisher model as the only process governing within-host iSNV frequency dynamics. Our results contribute to the growing number of studies that point towards the important role of genetic drift in shaping patterns of genetic diversity in IAV populations within acutely infected hosts. It further raises questions about whether and what other processes, such as spatial compartmentalization, viral progeny production dynamics with strong skew, or selection, may be needed to explain patterns of within-host IAV evolution.

evolutionary biology↗

Common misspecification of the generation interval leads to reproduction number underestimation in phylodynamic inference

Generation intervals are distributions that describe the time between infection and onward transmission. They are a key epidemiological quantity because, together with the reproduction number R, they determine the population-level growth rate of a pathogen and its doubling time. Conversely, when fitting epidemiological models to data, assumed generation intervals impact R inference. This is well-known from studies that have used case data for R inference, with many studies emphasizing the importance of choosing an accurate distribution for the generation interval. In phylodynamic inference of R, the generation interval distribution is often not explicitly mentioned, and the impact of generation interval misspecification has been studied less. Here, we explore the impact of a commonly assumed (but rarely empirically accurate) exponential generation interval distribution on the estimation of R in phylodynamic inference. Using simulations and inference on these simulated datasets, we find that during the early exponential growth of an epidemic, if the generation interval is assumed to be exponentially distributed when it actually has a lower variance, then estimates of R will be biased low. Furthermore, uncertainty in the biased R estimates will be small. Our work highlights the importance of acknowledging implicit generation interval assumptions in phylodynamic inference and points to the need for methodological developments in phylodynamic inference to provide greater flexibility in the specification of accurate generation intervals.

evolutionary biology↗

Immune Pressure is Key to Understanding Observed Patterns of Respiratory Virus Evolution in Prolonged Infections

For a number of viral pathogens, prolonged infections have been hypothesized to be the source of new variants that emerge and spread at the level of the host population. This potential role of prolonged infections has been highlighted most recently in the context of SARS-CoV-2, with variants of concern Alpha and Omicron likely to have evolved in immunocompromised individuals experiencing long-term viral infections. Analyses of sequenced viral samples from prolonged infections have indicated that there are several consistent patterns of evolution observed across these infections. These patterns include accelerated rates of nonsynonymous substitution, viral genetic diversification into distinct lineages, parallel substitutions across infected individuals, and heterogeneity in rates of antigenic evolution. Here, we use within-host model simulations to explore the drivers of these evolutionary patterns. Our simulations build on a tunably rugged fitness landscape model to first assess the role that mutations that impact only viral replicative fitness have in driving these patterns. They then further incorporate pleiotropic sites that jointly impact replicative fitness and antigenicity to assess the role that immune pressure has on these patterns. Through simulation, we find that the empirically observed patterns of viral evolution in prolonged infections cannot be robustly explained by viral populations evolving on replicative fitness landscapes alone. Instead, we find that immune pressure is needed to consistently reproduce the observed patterns. While our simulation models were designed to shed light on drivers of viral evolution in prolonged infections with respiratory viruses that generally cause acute infection, their structure can be used to better understand viral evolution in other acutely-infecting viruses such as noroviruses that can cause prolonged infection as well as viruses such as HIV that are known to chronically infect.

evolutionary biology↗

Quantifying viral pandemic potential from experimental transmission studies

In an effort to avert future pandemics, surveillance studies aimed at identifying zoonotic viruses at high risk of spilling over into humans act to monitor the viral chatter at the animal-human interface. These studies are hampered, however, by the diversity of zoonotic viruses and the limited tools available to assess pandemic risk. Methods currently in use include the characterization of candidate viruses using in vitro laboratory assays and experimental transmission studies in animal models. However, transmission experiments yield relatively low-resolution outputs that are not immediately translatable to projections of viral dynamics at the level of a host population. To address this gap, we present an analytical framework to extend the use of measurements from experimental transmission studies to generate more quantitative risk assessments. Specifically, we use within-host viral titer data from index and contact animals to estimate parameters relevant to transmission, including an estimate of transmissibility. We then extended this model to estimate epidemiological parameters, such as the basic reproductive number and generation interval. To illustrate these approaches, we present them in the context of two influenza A virus (IAV) ferret transmission experiments: one using influenza A/California/07/2009 (Cal/2009) and one using influenza A/Hong Kong/1/1968 (Hong Kong/1968). Despite estimating broadly similar transmissibilities for Cal/2009 and Hong Kong/1968, we conclude that Cal/2009 has a higher pandemic potential. This difference in pandemic potential seems to be primarily driven by its more robust within-host replication. Our results critically depend on several assumptions, namely that the within-host viral dynamics in humans and those in the model animal used (here, ferrets) share quantitative similarities and that viral transmissibility between model animals reflects viral transmissibility between humans. The methods we present to assess relative pandemic risk across viral isolates can be used to improve quantitative risk assessment of other emerging viruses of pandemic concern. Author summaryPandemic viruses often originate in animal reservoirs. Surveillance studies at the animal-human interface try to identify which of the many viruses circulating in animals could cause a pandemic in humans. Experimental transmission studies in animal models are one such method used to characterize potential pandemic viruses. However, only simple outcomes usually get reported from these experiments, such as the fraction of animals that become infected in such an experiment. Here, we develop a framework to extract more information from these experiments. Specifically, we use data on the virus population within infected ferrets to extrapolate information about transmission. We then simulate many transmission events to estimate epidemiological parameters commonly used in public health to quantify pandemic potential. Our methods therefore allow us to use commonly gathered data from experimental transmission studies to estimate epidemic dynamics in humans. This set of methods can be used to compare the relative pandemic risk of animal viruses before they emerge in humans.

microbiology↗

ScTree: Scalable and robust mechanistic integration of epidemiological and genomic data for transmission tree inference

Phylodynamic models capture joint epidemiological-evolutionary dynamics during an outbreak, providing a powerful tool to enhance understanding and management of disease transmission. Existing phylodynamic approaches, however, mostly rely on various non-mechanistic or semi-mechanistic approximations of the underlying epidemiological-evolutionary process. Previous work by Lau et al. [1] has shown that full Bayesian mechanistic models, without relying on these approximations, can enable highly accurate joint inference of the epidemiological-evolutionary dynamics including the unobserved transmission tree [1, 2]. However, the Lau method faces major computational bottlenecks. As the volume of genomic data collected during outbreaks continues to grow, it is crucial to develop scalable yet accurate phylodynamic methods. Here we propose a new Bayesian phylodynamic model, overcoming the major scalability issue in the Lau 2015 method and enabling a readily deployable, yet accurate, phylodynamic modeling framework. Specifically, we develop a scalable spatio-temporal phylodynamic framework for inferring the transmission tree (ScTree) and other key epidemiological parameters considering the infinite sites assumption in modeling mutation on the sequence level, in contrast to Lau 2015 in which mutation was modeled explicitly on the nucleotide level. Our approach features full Bayesian implementation utilizing a realistic likelihood to mechanistically integrate epidemiological and evolutionary processes. We develop a computationally-efficient data-augmentation Markov Chain Monte Carlo algorithm, inferring key model parameters and unobserved dynamics including the transmission tree. We assess performance of our method using multiple simulated outbreak datasets. Our results indicate that our method can achieve high inference accuracy, comparable to the performance of the Lau 2015 method. Additionally, our method scales significantly more efficiently for large outbreaks, with computing time increasing linearly with outbreak size, compared to the exponential scaling of the Lau method. We also demonstrate our methods utility by applying our validated modeling framework to a dataset describing a foot-and-mouth disease outbreak in the UK [3]. Our results show that our method is able to generate estimates of the transmission dynamics consistent with those from the Lau 2015 method, further demonstrating the robustness of our new approach. In summary, our method provides a computationally-efficient, highly scalable, accurate modeling framework for inferring the joint spatio-temporal dynamics of epidemiological and evolutionary processes, facilitating timely and effective outbreak responses in space and time. Our method is implemented in our R package ScTree. Author summaryPhylodynamic models integrate epidemiological and evolutionary dynamics to better understand disease transmission during outbreaks. However, many existing models rely on approximations that limit their accuracy and interpretability. Previous work by Lau et al. has shown that full Bayesian mechanistic models, without relying on these approximations, can enable highly accurate joint inference of the epidemiological-evolutionary dynamics including the unobserved transmission tree. Their method, however, faced significant computational challenges, particularly with large datasets. In this study, we present a new Bayesian phylodynamic model, ScTree, designed to overcome the scalability issues of the Lau 2015 method. By adopting the infinite-sites assumption for modeling mutations, rather than explicitly modeling nucleotide-level changes, ScTree achieves a significant improvement in computational efficiency while retaining accuracy of model inference. Our method, validated through simulations and real outbreak data, provides results comparable to the original Lau model but at a fraction of the computational cost, demonstrating its scalability and practical application for real-time outbreak responses. ScTree is implemented as an R package, making it accessible for further research and public health use.

evolutionary biology↗

Dispersal of influenza virus populations within the respiratory tract shapes their evolutionary potential

Viral infections are characterized by dispersal from an initial site to secondary locations within the host. How the resultant spatial heterogeneity shapes within-host genetic diversity and viral evolutionary pathways is poorly understood. Here we show that dispersal within and between the nasal cavity and trachea maintains diversity and is therefore conducive to adaptive evolution, whereas dispersal to the lungs is stochastic and gives rise to population heterogeneity. We infected ferrets either intranasally or by aerosol with a barcoded influenza A/California/07/2009 (H1N1) virus. At 1, 2 or 4 days post infection, dispersal was assessed by collecting 52 samples from throughout the respiratory tract of each animal. Irrespective of inoculation route, barcode compositions across the nasal turbinates and trachea were similar and highly diverse, revealing little constraint on the establishment of infection in the nasal cavity and descent through the trachea. By comparison, infection of the lungs produced genetically distinct viral populations. Lung populations were pauci-clonal, suggesting that each seeded location received relatively few viral genotypes. While aerosol inoculation gave distinct populations at every lung site sampled, within-host dispersal after intranasal inoculation produced larger patches, indicative of local expansion following seeding of the lungs. Throughout the respiratory tract, barcode diversity declined over time, but new diversity was generated through mutation. De novo variants were often unique to a given location, indicating that localized replication following dispersal resulted in population divergence. In summary, dispersal within the respiratory tract operates differently between regions and contributes to the potential for viral evolution to proceed independently in multiple within-host subpopulations.

microbiology↗

Phylodynamics beyond neutrality: The impact ofincomplete purifying selection on viral phylogeniesand inference

Viral phylodynamics focuses on using sequence data to make inferences about the population dynamics of viral infectious diseases. These inferences commonly include estimation of the viral growth rate, the reproduction number, and the time of most recent common ancestor. With few exceptions, existing phylodynamic inference approaches assume that all observed and ancestral viral genetic variation is fitness-neutral. This assumption is violated more often than not, with a large body of analyses indicating that fitness varies substantially among genotypes circulating viral populations. Here, we focus specifically on fitness variation arising from deleterious mutations, asking whether incomplete purifying selection of deleterious mutations has the potential to bias phylodynamic inference. We use simulations of an exponentially growing population to explore how incomplete purifying selection distorts tree shape as well as how it shifts the distribution of non-neutral mutations over trees. Consistent with previous results, we find that incomplete purifying selection strongly shapes the distribution of mutations while only weakly impacting tree shape. Despite incomplete purifying selection shifting the distribution of mutations, we find little discernible bias in estimates of the viral growth rate and times of the most recent common ancestor. Our results reassuringly indicate that existing phylodynamic inference approaches may not yield biased epidemiological parameter estimates in the face of incomplete purifying selection, although more work is needed to assess the generalizability of these findings.

evolutionary biology↗

Waning immunity drives respiratory virus evolution and reinfection

Reinfections with respiratory viruses such as influenza viruses and coronaviruses are thought to be driven by ongoing antigenic immune escape in the viral population. However, this does not explain why antigenic variation is frequently observed in these viruses relative to viruses such as measles that undergo systemic replication. Here, we suggest that the rapid rate of waning immunity in the respiratory tract is the key driver of antigenic evolution in respiratory viruses. Waning immunity results in hosts with immunity levels that protect against homologous reinfection but are insufficient to protect against infection with a heterologous, antigenically different strain. As such, when partially immune hosts are present at a high enough density, an immune escape variant can invade the viral population even though that variant cannot infect fully immune hosts. Invasion can occur even when the variants immune escape mutation incurs a fitness cost, and we expect the expanding mutant population will evolve compensatory mutations that mitigate this cost. Thus the mutant lineage should replace the wild-type, and as immunity to it builds, the process will repeat. Our model provides a new explanation for the pattern of successive emergence and replacement of antigenic variants that has been observed in many respiratory viruses. We discuss our model relative to others for understanding the drivers of antigenic evolution in these and other respiratory viruses.

evolutionary biology↗

Genetic drift and purifying selection shape within-host influenza A virus populations during natural swine infections

Patterns of within-host influenza A virus (IAV) diversity and evolution have been described in natural human infections, but these patterns remain poorly characterized in non-human hosts. Elucidating these dynamics is important to better understand IAV biology and the evolutionary processes that govern spillover into humans. Here, we sampled an IAV outbreak in pigs during a week-long county fair to characterize viral diversity and evolution in this important reservoir host. Nasal wipes were collected on a daily basis from all pigs present at the fair, yielding up to 421 samples per day. Subtyping of PCR-positive samples revealed the co-circulation of H1N1 and H3N2 subtype IAVs. PCR-positive samples with robust Ct values were deep-sequenced, yielding 506 sequenced samples from a total of 253 pigs. Based on higher-depth re-sequenced data from a subset of these initially sequenced samples (260 samples from 168 pigs), we characterized patterns of within-host IAV genetic diversity and evolution. We find that IAV genetic diversity in single-subtype infected pigs is low, with the majority of intra-host single nucleotide variants (iSNVs) present at frequencies of <10%. The ratio of the number of nonsynonymous to the number of synonymous iSNVs is significantly lower than under the neutral expectation, indicating that purifying selection shapes patterns of within-host viral diversity in swine. The dynamic turnover of iSNVs and their pronounced frequency changes further indicate that genetic drift also plays an important role in shaping IAV populations within pigs. Taken together, our results highlight similarities in patterns of IAV genetic diversity and evolution between humans and swine, including the role of stochastic processes in shaping within-host IAV dynamics.

evolutionary biology↗

Transmission bottleneck size estimation from de novo viral genetic variation

Sequencing of viral infections has become increasingly common over the last decade. Deep sequencing data in particular have proven useful in characterizing the roles that genetic drift and natural selection play in shaping within-host viral populations. They have also been used to estimate transmission bottleneck sizes from identified donor-recipient pairs. These bottleneck sizes quantify the number of viral particles that establish genetic lineages in the recipient host and are important to estimate due to their impact on viral evolution. Current approaches for estimating bottleneck sizes exclusively consider the subset of viral sites that are observed as polymorphic in the donor individual. However, allele frequencies can change dramatically over the course of an individuals infection, such that sites that are polymorphic in the donor at the time of transmission may not be polymorphic in the donor at the time of sampling and allele frequencies at donor-polymorphic sites may change dramatically over the course of a recipients infection. Because of this, transmission bottleneck sizes estimated using allele frequencies observed at a donors polymorphic sites may be considerable underestimates of true bottleneck sizes. Here, we present a new statistical approach for instead estimating bottleneck sizes using patterns of viral genetic variation that arose de novo within a recipient individual. Specifically, our approach makes use of the number of clonal viral variants observed in a transmission pair, defined as the number of viral sites that are monomorphic in both the donor and the recipient but carry different alleles. We first test our approach on a simulated dataset and then apply it to both influenza A virus sequence data and SARS-CoV-2 sequence data from identified transmission pairs. Our results confirm the existence of extremely tight transmission bottlenecks for these two respiratory viruses, using an approach that does not tend to underestimate transmission bottleneck sizes.

evolutionary biology↗

Influenza A genomic diversity during human infections underscores the strength of genetic drift and the existence of tight transmission bottlenecks

Influenza infections result in considerable public health and economic impacts each year. One of the contributing factors to the high annual incidence of human influenza is the viruss ability to evade acquired immunity through continual antigenic evolution. Understanding the evolutionary forces that act within and between hosts is therefore critical to interpreting past trends in influenza virus evolution and in predicting future ones. Several studies have analyzed the longitudinal patterns of influenza A virus genetic diversity in natural human infections to assess the relative contributions of selection and genetic drift on within-host evolution. However, in these natural infections, within-host viral populations harbor very few single nucleotide variants, limiting our resolution in understanding the forces acting on these populations in vivo. Further, low levels of within host viral genetic diversity limit the ability to infer the extent of drift across transmission events. Here, we propose to use influenza virus genomic diversity as an alternative signal to better understand within and between host patterns of viral evolution. Specifically, we focus on the dynamics of defective viral genomes (DVGs) which harbor large internal deletions in one or more of influenza viruss eight gene segments. Our longitudinal analyses of DVGs show that influenza A virus populations are highly dynamic within hosts, corroborating previous findings based on viral genetic diversity that point towards the importance of genetic drift in driving within-host viral evolution. Further, our analysis of DVG populations across transmission pairs indicate that DVGs rarely appeared to be shared, consistent with previous findings indicating the presence of tight transmission bottlenecks. Our analyses demonstrate that viral genomic diversity can be used to complement analyses based on viral genetic diversity to reveal processes that drive viral evolution within and between hosts. Author summaryDuring viral replication within infected individuals, different types of mutation can occur. Point mutations introduce genetic diversity, with some sites in the viral genome becoming polymorphic at the viral population level. Structural mutations that result in deletions in the viral genome can also occur, contributing to genomic diversity in the viral population. Defective viral genomes (DVGs) are a particular type of this genomic diversity that are incapable of establishing productive infection on their own, but persist during infection through cellular coinfection with infectious virus. Here, we first identify DVGs in deep sequencing data from natural infections of influenza A virus subtype H3N2 and then use these DVGs to inform our understanding of within-host viral evolution and transmission dynamics. By analyzing DVG populations over time within infected individuals and between transmission pairs, we find that influenza A virus populations are shaped by genetic drift and tight transmission bottlenecks. Our results add to our understanding of influenza genomic diversity within-and between-hosts and demonstrate the possibility of considering the full suite of viral diversity to understand drivers of viral evolution.

evolutionary biology↗

Within-host evolutionary dynamics and tissue compartmentalization during acute SARS-CoV-2 infection

The global evolution of SARS-CoV-2 depends in part upon the evolutionary dynamics within individual hosts with varying immune histories. To characterize the within-host evolution of acute SARS-CoV-2 infection, we deep sequenced saliva and nasal samples collected daily from immune and unvaccinated individuals early during infection. We show that longitudinal sampling facilitates high-confidence genetic variant detection and reveals evolutionary dynamics missed by less-frequent sampling strategies. Within-host dynamics in both naive and immune individuals appeared largely stochastic; however, we identified clear mutational hotspots within the viral genome, consistent with selection and differing between naive and immune individuals. In rare cases, minor genetic variants emerged to frequencies sufficient for forward transmission. Finally, we detected significant genetic compartmentalization of virus between saliva and nasal swab sample sites in many individuals. Altogether, these data provide a high-resolution profile of within-host SARS-CoV-2 evolutionary dynamics.

microbiology↗

Influenza A virus reassortment in mammals gives rise to genetically distinct within-host sub-populations

Influenza A virus (IAV) genetic exchange through reassortment has the potential to accelerate viral evolution and has played a critical role in the generation of multiple pandemic strains. For reassortment to occur, distinct viruses must co-infect the same cell. The spatio-temporal dynamics of viral dissemination within an infected host therefore define opportunity for reassortment. Here, we used wild type and synonymously barcoded variant viruses of a pandemic H1N1 strain to examine the within-host viral dynamics that govern reassortment in guinea pigs, ferrets and swine. The first two species are well-established models of human influenza, while swine are a natural host and a frequent conduit for cross-species transmission and reassortment. Our results show reassortment to be pervasive in all three hosts but less frequent in swine than in ferrets and guinea pigs. In ferrets, tissue-specific differences in the opportunity for reassortment are also evident, with more reassortants detected in the nasal tract than the lower respiratory tract. While temporal trends in viral diversity are limited, spatial patterns are clear, with heterogeneity in the viral genotypes detected at distinct anatomical sites revealing extensive compartmentalization of reassortment and replication. Our data indicate that the dynamics of viral replication in mammals allow diversification through reassortment but that the spatial compartmentalization of variants likely shapes their evolution and onward transmission.

microbiology↗