Viral evolution

The reticuloendotheliosis viruses (REVs) comprise several closely related amphotropic retroviruses isolated from birds. These viruses exhibit several highly unusual characteristics that have not so far been adequately explained, including their extremely close relationship to mammalian retroviruses, and their presence as endogenous sequences within the genomes of certain large DNA viruses. We present evidence for an iatrogenic origin of REVs that accounts for these phenomena. Firstly, we identify endogenous retroviral fossils in mammalian genomes that share a unique recombinant structure with REVs—unequivocally demonstrating that REVs derive directly from mammalian retroviruses. Secondly, through sequencing of archived REV isolates, we confirm that contaminated Plasmodium lophurae stocks have been the source of multiple REV outbreaks in experimentally infected birds. Finally, we show that both phylogenetic and historical evidence support a scenario wherein REVs originated as mammalian retroviruses that were accidentally introduced into avian hosts in the late 1930s, during experimental studies of P. lophurae, and subsequently integrated into the fowlpox virus (FWPV) and gallid herpesvirus type 2 (GHV-2) genomes, generating recombinant DNA viruses that now circulate in wild birds and poultry. Our findings provide a novel perspective on the origin and evolution of REV, and indicate that horizontal gene transfer between virus families can expand the impact of iatrogenic transmission events.

Primate lentiviruses, including the human and simian immunodeficiency viruses (HIV and SIV), produce infections marked by persistent, ongoing viral replica-tion. This occurs despite the presence of virus-specific adaptive immune responses, including antibodies targeting the viral envelope glycoprotein (Env), and evolution of antibody-escape variants is a well-documented feature of lentiviral infection. Here, we examined the evolutionary dynamics of the SIV env gene during early infection (29 weeks postinfection) in a cohort of four SIV mac 251-infected rhesus macaques. We tracked env evolution during acute and early infection using frequent sampling and ultradeep sequencing of viral populations, capturing a transmission bottleneck and the subsequent reestablishment of Env diversity. A majority of changes in the gp120 subunit mapped to two short clusters, one in the first variable region (V1) and one in V4, while most changes in the gp41 subunit appeared in the cytoplasmic domain. Variation in V1 was dominated by short duplications and deletions of repetitive sequence, while variation in V4 was marked by short in-frame deletions and closely overlapping substitutions. The most common substitutions in both patches did not alter viral replicative fitness when tested using a highly sensitive, deep-sequencing-based competition assay. Our results, together with the observation that very similar or identical patterns of sequence evolution also occur in different macaque species infected with related but divergent strains of SIV, suggest that resistance to early, strain-specific anti-Env antibodies is the result of temporally and mutationally predictable pathways of escape that occur during the early stages of infection. IMPORTANCE The envelope glycoprotein (Env) of primate lentiviruses mediates entry by binding to host cell receptors followed by fusion of the viral membrane with the cell membrane. The exposure of Env complexes on the surface of the virion results in targeting by antibodies, leading to selection for virus escape mutations. We used the SIV/rhesus macaque model to track in vivo evolution of variation in Env during acute/early infection in animals with and without antibody responses to Env, uncovering remarkable variation in animals with antibody responses within weeks of infection. Using a deep-sequencing-based fitness assay, we found substitutions associated with antibody escape had little to no effect on inherent replicative capacity. The ability to readily propagate advantageous changes that incur little to no replicative fitness costs may be a mechanism to maintain continuous replication under constant immune selection, allowing the virus to persist for months to years in the infected host.

We introduce VERSO, a two-step framework for the characterization of viral evolution from sequencing data of viral genomes, which improves over phylogenomic approaches for consensus sequences. VERSO exploits an efficient algorithmic strategy to return robust phylogenies from clonal variant profiles, also in conditions of sampling limitations. It then leverages variant frequency patterns to characterize the intra-host genomic diversity of samples, revealing undetected infection chains and pinpointing variants likely involved in homoplasies. On simulations, VERSO outperforms state-of-the-art tools for phylogenetic inference. Notably, the application to 6726 Amplicon and RNA-seq samples refines the estimation of SARS-CoV-2 evolution, while co-occurrence patterns of minor variants unveil undetected infection paths, which are validated with contact tracing data. Finally, the analysis of SARS-CoV-2 mutational landscape uncovers a temporal increase of overall genomic diversity, and highlights variants transiting from minor to clonal state and homoplastic variants, some of which falling on the spike gene. Available at: https://github.com/BIMIB-DISCo/VERSO.

In the present work we analyze the problem of adaptation and evolution of RNA virus populations, by defining the basic stochastic model as a multivariate branching process in close relation with the branching process advanced by Demetrius, Schuster and Sigmund (" Polynucleotide evolution and branching processes " , Bull. Math. Biol. 46 (1985) 239-262), in their study of polynucleotide evolution. We show that in the absence of beneficial forces the model is exactly solvable. As a result it is possible to prove several key results directly related to known typical properties of these systems like (i) proof, in the context of the theory of branching processes, of the lethal mutagenesis criterion proposed by Bull, Sanjuán and Wilke (" Theory of lethal mutagenesis for viruses " , J. Virology 18 (2007) 2930-2939); (ii) a new proposal for the notion of relaxation time with a quantitative prescription for its evaluation and (iii) the quantitative description of the evolution of the expected values in four distinct regimes: transient, " stationary " equilibrium, extinction threshold and lethal mutagenesis. Moreover, new insights on the dynamics of evolving virus populations can be foreseen.

Hepatitis C virus (HCV) has infected over 170 million
people worldwide and creates a huge disease burden
due to chronic, progressive liver disease. HCV is a single-stranded, positive sense, RNA virus, member of the
Flaviviridae family. The high error rate of RNA-dependent
RNA polymerase and the pressure exerted by the host
immune system, has driven the evolution of HCV into
7 different genotypes and more than 67 subtypes. HCV
evolves by means of different mechanisms of genetic
variation. On the one hand, its high mutation rates
generate the production of a large number of different
but closely related viral variants during infection, usually
referred to as a quasispecies. The great quasispecies
variability of HCV has also therapeutic implications since
the continuous generation and selection of resistant or
fitter variants within the quasispecies spectrum might
allow viruses to escape control by antiviral drugs. On
the other hand HCV exploits recombination to ensure its
survival. This enormous viral diversity together with some
host factors has made it difficult to control viral dispersal.
Current treatment options involve pegylated interferon-α
and ribavirin as dual therapy or in combination with a
direct-acting antiviral drug, depending on the country.
Despite all the efforts put into antiviral therapy studies,
eradication of the virus or the development of a preventive
vaccine has been unsuccessful so far. This review focuses
on current available data reported to date on the genetic
mechanisms driving the molecular evolution of HCV
populations and its relation with the antiviral therapies
designed to control HCV infection.