Biology subjects

Tesla, B.

Publications and source records attributed to Tesla, B..

7 recordsLinked to original sources

bioRxiv2023

Understanding off-target growth defects introduced to influenza A virus by synonymous recoding

CpG dinucleotides are under-represented in the genomes of most RNA viruses. Synonymously increasing CpG content of a range of RNA viruses reliably causes replication defects due to the recognition of CpG motifs in RNA by cellular Zinc-finger Antiviral Protein (ZAP). Prior to the discovery of ZAP as a CpG sensor, we described an engineered influenza A virus (IAV) enriched for CpGs in segment 5 that displays the expected replication defects. However, we report here that this CpG-high ( CpGH) mutant is not attenuated by ZAP. To understand this, we sought to uncover the alternative attenuation mechanism(s). IAV segment 5 encodes NP, a component of the viral RNA replication complex. Unexpectedly, while CpG enrichment resulted in depleted segment 5 transcript and NP protein abundance, this did not impair viral polymerase activity. A pair of nucleotide changes, introduced as compensatory changes to maintain base frequencies, were instead found to be responsible for the replication defect. These mutations resulted in the encoding of a stretch of eight consecutive adenosines (8A), a phenomenon not seen in natural IAV isolates. Sequencing experiments revealed evidence of viral polymerase slippage at this site, resulting in the production of aberrant peptides and type I interferon induction. When the nucleotides in either of these two positions were restored to wildtype sequence, no viral attenuation was seen, despite the 86 extra CpGs encoded by this virus. Conversely, when these two adenosines were introduced into wildtype virus (thereby introducing the 8A tract), viral attenuation, polymerase slippage, aberrant peptide production and type I interferon induction were apparent. That a single nucleotide change can offset the growth defects in a virus designed to have a formidable barrier to wild-type reversion highlights the importance of understanding the processes underlying viral attenuation. The lessons from this study will inform improved recoding designs in the future.

microbiology↗

bioRxiv2021

Cool temperatures limit Zika virus genome replication

Zika virus is a mosquito-borne flavivirus known to cause severe birth defects and neuroimmunological disorders. We have previously demonstrated that mosquito transmission of Zika virus decreases with temperature. While transmission was optimized at 29{degrees}C, it was limited at cool temperatures (< 22{degrees}C) due to poor virus establishment in the mosquitoes. Temperature is one of the strongest drivers of vector-borne disease transmission due to its profound effect on ectothermic mosquito vectors, viruses, and their interaction. Although there is substantial evidence of temperature effects on arbovirus replication and dissemination inside mosquitoes, little is known about whether temperature affects virus replication directly or indirectly through mosquito physiology. In order to determine the mechanisms behind temperature-induced changes in Zika virus transmission potential, we investigated different steps of the virus replication cycle in mosquito cells (C6/36) at optimal (28{degrees}C) and cool (20{degrees}C) temperatures. We found that cool temperature did not alter Zika virus entry or translation but reduced the amount of double-stranded RNA replication intermediates. If replication complexes were first formed at 28{degrees}C and the cells were subsequently shifted to 20{degrees}C, the late steps in the virus replication cycle were efficiently completed. These data suggest that cool temperature decreases the efficiency of Zika virus genome replication in mosquito cells. This phenotype was observed in the Asian-lineage of Zika virus, while the African-lineage Zika virus was less restrictive at 20{degrees}C. ImportanceWith half of the human population at risk, arboviral diseases represent a substantial global health burden. Zika virus, previously known to cause sporadic infections in humans, emerged in the Americas in 2015 and quickly spread worldwide. There was an urgent need to better understand the disease pathogenesis, develop therapeutics and vaccines, as well as to understand, predict, and control virus transmission. In order to efficiently predict the seasonality and geography for Zika virus transmission, we need a deeper understanding of the host-pathogen interactions and how they can be altered by environmental factors such as temperature. Identifying the step in the virus replication cycle that is inhibited in cool conditions can have implications in modeling the temperature suitability for arbovirus transmission as global environmental patterns change. Understanding the link between pathogen replication and environmental conditions can potentially be exploited to develop new vector control strategies in the future.

microbiology↗

bioRxiv2019

Effects of temperature on Zika dynamics and control

ABSTRACTWhen a formerly rare pathogen emerges to cause a pandemic, it is critical to understand the ecology of the disease dynamics and its potential effects on disease control. Here, we take advantage of newly available experimental data to parameterize a temperature-dependent dynamical model of Zika virus (ZIKV) transmission, and analyze the effects of temperature variability and the parameters related to control strategies on ZIKV R0 and the final epidemic size (i.e., total number of human cases). Sensitivity analyses identified that R0 and the final epidemic size were largely driven by different parameters, with the exception of temperature, which is the dominant driver of epidemic dynamics in the models. Our estimate of R0 had a single optimum temperature (≈ 30° C), comparable to recently published results (≈ 29°)1. However, the total number of human cases (“final epidemic size”) is maximized across a wider temperature range, from 24 to 36°C. The models indicate that the disease is highly sensitive to seasonal temperature variation. For example, although the model predicts that Zika transmission cannot occur at a constant temperature of 22°C, with seasonal variation of 5°C around a mean of 22°C, the model predicts a larger epidemic than what would occur at a constant 30°C, the temperature predicted to maximize R0. This suggests that the potential geographic range of Zika is wider than indicated from static R0 models, underscoring the importance of climate dynamics and variation on emerging infectious diseases.Competing Interest StatementThe authors have declared no competing interest.View Full Text

ecology↗

bioRxiv2019

Temperature Dramatically Shapes Mosquito Gene Expression with Consequences for Mosquito-Zika Virus Interactions

Vector-borne flaviviruses are emerging threats to human health. For successful transmission, the virus needs to efficiently enter mosquito cells, replicate within, and escape several tissue barriers while mosquitoes elicit major transcriptional responses to flavivirus infection. This process will not only be affected by the specific mosquito-pathogen pairing, but also variation in key environmental variables such as temperature. Thus far, few studies have examined the molecular responses triggered by temperature and how these responses modify infection outcomes despite substantial evidence showing strong relationships between temperature and transmission in a diversity of systems. To define the host transcriptional changes associated with temperature variation during the early infection process, we compared the transcriptome of mosquito midgut samples from mosquitoes exposed to Zika virus (ZIKV) and non-exposed mosquitoes housed at three different temperatures (20, 28, and 36{degrees}C). While the high temperature samples did not have significant changes from standard rearing conditions (28{degrees}C) 48 hr post-exposure, the transcriptome profile of mosquitoes housed at 20{degrees}C was dramatically different. The expression of genes most altered by the cooler temperature involved aspects of blood-meal digestion, ROS metabolism, and mosquito innate immunity. Further, we did not find significant differences in the viral RNA copy number between 24 and 48 hr post-exposure at 20{degrees}C, suggesting ZIKV replication is limited by cold-induced changes to the mosquito midgut environment. In ZIKV-exposed mosquitoes, vitellogenin, a lipid carrier protein, was the most up-regulated at 20{degrees}C. Our results provide a deeper understanding of the temperature-triggered transcriptional changes in Aedes aegypti and can be used to further define the molecular mechanisms driven by environmental temperature variation.\n\nContribution to the Field StatementA variety of methods for engineering refractory mosquitoes are currently being studied and show promise for disease control. Although considerable effort has been put into understanding the immune system of mosquitoes in response to infections, almost nothing is understood about environmental influences in regulating these responses. Here, we used RNA sequencing to study the effect of temperature on the mosquito transcriptome profile, as well as assess the changes in the immune response to ZIKV infection at three different temperatures. We found a remarkable effect of temperature on the transcriptome profile of mosquitoes exposed to cool conditions (20{degrees}C) after imbibing a blood meal, as well as accumulation of transcripts involved with different mechanisms associated with blood meal digestion, metabolism, and some components of the immune response in mosquitoes. Our results provide new insights in potential mechanisms that limit temperature-driven pathogen establishment and replication within the mosquito vector.

molecular biology↗

bioRxiv2018

Impacts of temperature on Zika virus transmission potential: combining empirical and mechanistic modeling approaches

Temperature is a strong driver of vector-borne disease transmission. Yet, for emerging arboviruses we lack fundamental knowledge on the relationship between transmission and temperature. Current models rely on the untested assumption that Zika virus responds similarly to dengue virus, potentially limiting our ability to accurately predict the spread of Zika. We conducted experiments to estimate the thermal performance of Zika virus (ZIKV) in field-derived Aedes aegypti across eight constant temperatures. We observed strong, unimodal effects of temperature on vector competence, extrinsic incubation period, and mosquito survival. We used thermal responses of these traits to update an existing temperature-dependent model to infer temperature effects on ZIKV transmission. ZIKV transmission was optimized at 29{degrees}C, and had a thermal range of 22.7{degrees}C - 34.7{degrees}C. Thus, as temperatures move toward the predicted thermal optimum (29{degrees}C) due to climate change, urbanization, or seasonally, Zika could expand north and into longer seasons. In contrast, areas that are near the thermal optimum were predicted to experience a decrease in overall environmental suitability. We also demonstrate that the predicted thermal minimum for Zika transmission is 5{degrees}C warmer than that of dengue, and current global estimates on the environmental suitability for Zika are greatly over-predicting its possible range.

ecology↗

bioRxiv2017

Estimating the effects of variation in viremia on mosquito susceptibility, infectiousness, and R0 of Zika in Aedes aegypti

Zika virus (ZIKV) is an arbovirus primarily transmitted by Aedes mosquitoes. Like most viral infections, ZIKV viremia varies over several orders of magnitude, with unknown consequences for transmission. To determine the effect of viral concentration on ZIKV transmission risk, we exposed field-derived Ae. aegypti mosquitoes to four doses (103, 104, 105, 106 PFU/mL) representative of potential variation in the field. We demonstrate that increasing ZIKV dose in the blood-meal significantly increases the probability of mosquitoes becoming infected and infectious, as well as the rate at which virus spreads to the saliva, but found no effect on dissemination efficiency or mosquito mortality. We also demonstrate that determining infection using RT-qPCR approaches rather than plaque assays potentially over-estimates key pathogen parameters, including the time at which mosquitoes become infectious and viral burden. Finally, using these data to parameterize an R0 model, we demonstrate that variation in viremia substantially affects transmission risk.

microbiology↗

bioRxiv2016

Fine-scale variation in microclimate across an urban landscape changes the capacity of Aedes albopictus to vector arbovirus

Most statistical and mechanistic models used to predict mosquito borne disease transmission incorporate climate drivers of disease transmission by utilizing environmental data collected at scales that are potentially coarser than what mosquito vectors actually experience. Temperature and relative humidity can vary greatly between indoor and outdoor environments, and can be influenced strongly by variation in landscape features. In the Aedes albopictus system, we conducted a proof-of-concept study in the vicinity of the University of Georgia to explore the effects of fine-scale microclimate variation on mosquito life history and vectorial capacity (VC). We placed Ae. albopictus larvae in artificial pots distributed across three replicate sites within three different land uses - urban, suburban, and rural, which were characterized by high, intermediate, and low proportions of impervious surfaces. Data loggers were placed into each larval environment and in nearby vegetation to record daily variation in water and ambient temperature and relative humidity. The number of adults emerging from each pot and their body size and sex were recorded daily. We found mosquito microclimate to significantly vary across the season as well as with land use. Urban sites were in general warmer and less humid than suburban and rural sites, translating into decreased larval survival, smaller body sizes, and lower per capita growth rates of mosquitoes on urban sites. Dengue transmission potential was predicted to be higher in the summer than the fall. Additionally, the effects of land use on dengue transmission potential varied by season. Warm summers resulted in a higher predicted VC on the cooler, rural sites, while warmer, urban sites had a higher predicted VC during the cooler fall season.

ecology↗