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Jacobs, N. T.

Publications and source records attributed to Jacobs, N. T..

5 recordsLinked to original sources

BlpC-mediated selfish program leads to rapid loss of Streptococcus pneumoniae clonal diversity during infection

Chromosomal barcoding and high-throughput sequencing were used to investigate the population dynamics of Streptococcus pneumoniae. During infant mouse colonization, >35-fold reduction in diversity and expansion of a single clonal lineage was observed within 1 day post-inoculation. This loss of diversity was not due to immune factors, host microbiota or exclusively because of genetic drift. Rather, it required the expression of blp bacteriocins induced by the BlpC-quorum sensing pheromone. This points towards the role of intra-strain competition whereby the subpopulation reaching a quorum eliminates others that have yet to activate the blp locus. We show that this loss of diversity also restricts the number of unique clones that could establish colonization during transmission between hosts. Moreover, we show that genetic variation in the blp locus is associated with transmissibility in the human population. We posit this is due to its importance in clonal selection and its role as a selfish element.

microbiology↗

Competitive exclusion strengthens selection for transmissibility and increases the benefit of recombination for within-host adaptation.

Pathogens experience selection at multiple scales, given the need to transmit between hosts and replicate within them. This presents the challenge of cross-scale selective conflict when adaptations to one scale compromise fitness at another, such as mutations that improve transmissibility but make individuals less competitive within hosts. Selection operates differently at these scales, with tight transmission bottlenecks subjecting pathogen populations to genetic drift, and large population sizes within hosts enabling efficient selection for beneficial mutations. Compounding the reduction in diversity by transmission bottlenecks is the occupant-intruder competitive strategy exhibited by some pathogens, where the first variant to colonize a host prevents later arriving variants from contributing to infection, preventing immigration and turning transmission into a "founder takes all" contest. Here, we used multiple modeling approaches to examine how this behavior affects the efficiency of selection for both transmissibility and within- host fitness. We find that in the face of a trade-off, selection for transmissibility is maximized under a tight transmission bottleneck that minimizes within-host competition during colonization. While mutations with increased within-host fitness are favored during within-host replication, an occupant-intruder strategy prevents these mutants from displacing established residents and propagating across the host population, leading to their extinction if they are insufficiently transmissible. Finally, a model of competition on the scale of the host population revealed that competitive exclusion limits the propagation of mutations with improved within-host fitness, unless resident populations can incorporate alleles from intruding variants by recombination. Thus, competitive exclusion may facilitate the improvement and maintenance of pathogen transmissibility, with directional recombination allowing resident populations to mitigate the potential loss of within-host fitness imposed by this occupant-intruder strategy. Author SummaryTransmission is a defining feature of infectious diseases, and so a better understanding of how transmissibility evolves is important for improving disease surveillance and prevention. Successful transmission is often achieved by a small number of individuals which, after establishing residency in a host, may prevent newcomers from participating in infection. Here, we use modeling to examine how competitive exclusion of challengers by resident populations affects the balance between within-host competitive ability and transmissibility. We find that competitive exclusion strengthens selection for transmissibility by disproportionately benefitting the first variant to colonize a host and preventing mutants that may be more competitive but less transmissible from displacing established residents. Competitive exclusion also limits the propagation of mutants that improve within-host fitness without reducing transmissibility, however, increasing the advantage of recombination that allows resident populations to acquire beneficial alleles from challengers. Competitive strategies that allow pathogens to "claim ownership" of hosts may thus help pathogen populations maintain transmissibility, with genetic recombination facilitating within-host adaptation through the incorporation of beneficial alleles from challengers.

evolutionary biology↗

Collective interactions augment influenza A virus replication in a host-dependent manner

Infection with a single influenza A virus (IAV) is only rarely sufficient to initiate productive infection. Here, we exploit both single-cell approaches and whole-animal systems to show that IAV reliance on multiple infection can form an important species barrier to infection. Namely, we find that H9N2 subtype viruses representative of those circulating widely at the poultry-human interface exhibit acute dependence on collective interactions in mammalian systems. This need for multiple infection is greatly reduced in the natural host. Quantification of incomplete viral genomes showed that their complementation accounts for the more moderate reliance on coinfection seen in avian cells, but not the added reliance seen in mammalian cells. This finding suggests an additional form of virus-virus interaction is needed to support infection in mammalian cells. Genetic mapping implicated the PA gene segment as a major driver of this phenotype and quantification of viral RNA synthesis indicated that both replication and transcription were affected. These findings indicate that multiple distinct mechanisms underlie IAV reliance on multiple infection and underscore the importance of virus-virus interactions in IAV infection, evolution and emergence.

microbiology↗

Dysregulation of M segment gene expression contributes to influenza A virus host restriction

The M segment of the 2009 pandemic influenza A virus (IAV) has been implicated in its emergence into human populations. To elucidate the genetic contributions of the M segment to host adaptation, and the underlying mechanisms, we examined a panel of isogenic viruses that carry avian- or human-derived M segments. Avian, but not human, M segments restricted viral growth and transmission in mammalian model systems, and the restricted growth correlated with increased expression of M2 relative to M1. M2 overexpression was associated with intracellular accumulation of autophagosomes, which was alleviated by interference of the viral proton channel activity by amantadine treatment. As M1 and M2 are expressed from the M mRNA through alternative splicing, we separated synonymous and non-synonymous changes that differentiate human and avian M segments and found that dysregulation of gene expression leading to M2 overexpression diminished replication, irrespective of amino acid composition of M1 or M2. Moreover, in spite of efficient replication, virus possessing a human M segment that expressed avian M2 protein at low level did not transmit efficiently. We conclude that (i) determinants of transmission reside in the IAV M2 protein, and that (ii) control of M segment gene expression is a critical aspect of IAV host adaptation needed to prevent M2-mediated dysregulation of vesicular homeostasis.\n\nAuthor summaryInfluenza A virus (IAV) pandemics arise when a virus adapted to a non-human host overcomes species barriers to successfully infect humans and sustain human-to-human transmission. To gauge the adaptive potential and therefore pandemic risk posed by a particular IAV, it is critical to understand the mechanisms underlying viral adaptation to human hosts. Here, we focused on the role of one of IAVs eight gene segments, the M segment, in host adaptation. Comparing the growth of IAVs with avian- and human-derived M segments in avian and mammalian systems revealed that the avian M segment restricts viral growth specifically in mammalian cells. We show that the mechanism underlying this host range phenotype is a dysregulation of viral gene expression when the avian IAV M segment is transcribed in mammalian cells. In particular, excess production of the M2 protein results in viral interference with cellular functions on which the virus relies. Our results therefore reveal that the use of cellular machinery to control viral gene expression leaves the virus vulnerable to over- or under-production of critical viral gene products in a new host species.

microbiology↗

Incomplete influenza A virus genomes are abundant but readily complemented during spatially structured viral spread

Viral genomes comprising multiple distinct RNA segments can undergo genetic exchange through reassortment, a process that facilitates viral evolution and can have major epidemiological consequences. Segmentation also allows the replication of incomplete viral genomes (IVGs), however, and evidence suggests that IVGs occur frequently for influenza A viruses. Here we quantified the frequency of IVGs using a novel single cell assay and then examined their implications for viral fitness. We found that each segment of influenza A/Panama/2007/99 (H3N2) virus has only a 58% probability of being present in a cell infected with a single virion. These observed frequencies accurately account for the abundant reassortment seen in co-infection, and suggest that an average of 3.7 particles are required for replication of a full viral genome in a cell. This dependence on multiple infection is predicted to decrease infectivity and to slow viral propagation in a well-mixed system. Importantly, however, modeling of spatially structured viral growth predicted that the need for complementation is met more readily when secondary spread occurs locally. This expectation was supported by experimental infections in which the level spatial structure was manipulated. Furthermore, a virus engineered to be entirely dependent on co-infection to replicate in vivo was found to grow robustly in guinea pigs, suggesting that coinfection is sufficiently common in vivo to support propagation of IVGs. The infectivity of this mutant virus was, however, reduced 815-fold relative wild-type and the mutant virus did not transmit to contacts. Thus, while incomplete genomes augment reassortment and contribute to within-host spread, the existence of rare complete IAV genomes may be critical for transmission to new hosts.

microbiology↗