skip to main content


Title: Activation of the autophagy pathway decreases dengue virus infection in Aedes aegypti cells
Abstract Background

Mosquito-borne dengue virus (DENV) causes major disease worldwide, impacting 50–100 million people every year, and is spread by the major mosquito vectorAedes aegypti. Understanding mosquito physiology, including antiviral mechanisms, and developing new control strategies have become an important step towards the elimination of DENV disease. In the study reported here, we focused on autophagy, a pathway suggested as having a positive influence on virus replication in humans, as a potential antiviral target in the mosquito.

Methods

To understand the role played by autophagy inAe. aegypti, we examined the activation of this pathway in Aag-2 cells, anAe. aegypti-derived cell line, infected with DENV. Rapamycin and 3-methyladenine, two small molecules that have been shown to affect the function of the autophagy pathway, were used to activate or suppress, respectively, the autophagy pathway.

Results

At 1-day post-DENV infection in Aag-2 cells, transcript levels of both the microtubule-associated protein light chain 3-phosphatidylethanolamine conjugate (LC3-II) and autophagy-related protein 1 (ATG1) increased. Rapamycin treatment activated the autophagy pathway as early as 1-h post-treatment, and the virus titer had decreased in the Aag-2 cells at 2 days post-infection; in contrast, the 3-methyladenine treatment did not significantly affect the DENV titer. Treatment with these small molecules also impacted the ATG12 transcript levels in DENV-infected cells.

Conclusions

Our studies revealed that activation of the autophagy pathway through rapamycin treatment altered DENV infection in the mosquito cells, suggesting that this pathway could be a possible antiviral mechanism in the mosquito system. Here we provide fundamental information needed to proceed with future experiments and to improve our understanding of the mosquito’s immune response against DENV.

Graphical Abstract 
more » « less
NSF-PAR ID:
10306103
Author(s) / Creator(s):
;
Publisher / Repository:
Springer Science + Business Media
Date Published:
Journal Name:
Parasites & Vectors
Volume:
14
Issue:
1
ISSN:
1756-3305
Format(s):
Medium: X
Sponsoring Org:
National Science Foundation
More Like this
  1. Abstract Background

    TheAedesaegyptimosquito is a vector of several viruses including dengue, chikungunya, zika, and yellow fever. Vector surveillance and control are the primary methods used for the control and prevention of disease transmission; however, public health institutions largely rely on measures of population abundance as a trigger for initiating control activities. Previous research found evidence that at the northern edge ofAe.aegypti’s geographic range, survival, rather than abundance, is likely to be the factor limiting disease transmission. In this study, we sought to test the utility of using body size as an entomological index to surveil changes in the age structure of field-collected femaleAedesaegypti.

    Methods

    We collected femaleAe.aegyptimosquitoes using BG sentinel traps in three cities at the northern edge of their geographic range. Collections took place during their active season over the course of 3 years. Female wing size was measured as an estimate of body size, and reproductive status was characterized by examining ovary tracheation. Chronological age was determined by measuring transcript abundance of an age-dependent gene. These data were then tested with female abundance at each site and weather data from the estimated larval development period and adulthood (1 week prior to capture). Two sources of weather data were tested to determine which was more appropriate for evaluating impacts on mosquito physiology. All variables were then used to parameterize structural equation models to predict age.

    Results

    In comparing city-specific NOAA weather data and site-specific data from HOBO remote temperature and humidity loggers, we found that HOBO data were more tightly associated with body size. This information is useful for justifying the cost of more precise weather monitoring when studying intra-population heterogeneity of eco-physiological factors. We found that body size itself was not significantly associated with age. Of all the variables measured, we found that best fitting model for age included temperature during development, body size, female abundance, and relative humidity in the 1 week prior to capture . The strength of models improved drastically when testing one city at a time, with Hermosillo (the only study city with seasonal dengue transmission) having the best fitting model for age. Despite our finding that there was a bias in the body size of mosquitoes collected alive from the BG sentinel traps that favored large females, there was still sufficient variation in the size of females collected alive to show that inclusion of this entomological indicator improved the predictive capacity of our models.

    Conclusions

    Inclusion of body size data increased the strength of weather-based models for age. Importantly, we found that variation in age was greater within cities than between cities, suggesting that modeling of age must be made on a city-by-city basis. These results contribute to efforts to use weather forecasts to predict changes in the probability of disease transmission by mosquito vectors.

    Graphical abstract 
    more » « less
  2. Abstract

    Mosquitoes are insects of interest because several species vector disease‐causing pathogens to humans and other vertebrates. We previously reported that mosquitoes from long‐term laboratory cultures require living bacteria in their gut to develop, but development does not depend on particular species of bacteria. Here, we focused on three distinct but interrelated areas of study to better understand the role of bacteria in mosquito development by studying field and laboratory populations ofAedes aegypti,Aedes albopictusandCulex quinquefasciatusfrom the southeastern United States. Sequence analysis of bacterial 16S rRNA gene amplicons showed that bacterial community composition differed substantially in larvae from different collection sites, whereas larvae from the same site shared similarities. Although previously unknown to be infected byWolbachia, results also indicated thatAe. aegyptifrom one field site hosted a dual infection. Regardless of collection site or factors likeWolbachiainfection, however, each mosquito species required living bacteria in their digestive tract to develop. Results also identified several concerns in using antibiotics to eliminate the bacterial community in larvae in order to study its developmental consequences. Altogether, our results indicate that several mosquito species require living bacteria for development. We also hypothesize these species do not rely on particular bacteria because larvae do not reliably encounter the same bacteria in the aquatic habitats they develop in.

     
    more » « less
  3. Abstract Background

    Effectively controlling heartworm disease—a major parasitic disease threatening animal health in the US and globally—requires understanding the local ecology of mosquito vectors involved in transmission. However, the key vector species in a given region are often unknown and challenging to identify. Here we investigate (i) the key vector species associated with transmission of the parasite,Dirofilaria immitis, in California and (ii) the climate and land cover drivers of vector presence.

    Methods

    To identify key mosquito vectors involved in transmission, we incorporated long-term, finely resolved mosquito surveillance data and dog heartworm case data in a statistical modeling approach (fixed-effects regression) that rigorously controls for other unobserved drivers of heartworm cases. We then used a flexible machine learning approach (gradient boosted machines) to identify the climate and land cover variables associated with the presence of each species.

    Results

    We found significant, regionally specific, positive associations between dog heartworm cases and the abundance of four vector species:Aedes aegypti(Central California),Ae. albopictus(Southern California),Ae. sierrensis(Central California), andCuliseta incidens(Northern and Central California). The proportion of developed land cover was one of the most important ecological variables predicting the presence or absence of the putative vector species.

    Conclusion

    Our results implicate three previously under-recognized vectors of dog heartworm transmission in California and indicate the land cover types in which each putative vector species is commonly found. Efforts to target these species could prioritize surveillance in these land cover types (e.g. near human dwellings in less urbanized settings forAe. albopictusandCs. incidens) but further investigation on the natural infection prevalence and host-biting rates of these species, as well as the other local vectors, is needed.

    Graphical Abstract 
    more » « less
  4. Sugden, Bill (Ed.)
    The scientific community is focused on developing antiviral therapies to mitigate the impacts of the ongoing novel coronavirus disease 2019 (COVID-19) outbreak. This will be facilitated by improved understanding of viral dynamics within infected hosts. Here, using a mathematical model in combination with published viral load data, we compare within-host viral dynamics of SARS-CoV-2 with analogous dynamics of MERS-CoV and SARS-CoV. Our quantitative analyses using a mathematical model revealed that the within-host reproduction number at symptom onset of SARS-CoV-2 was statistically significantly larger than that of MERS-CoV and similar to that of SARS-CoV. In addition, the time from symptom onset to the viral load peak for SARS-CoV-2 infection was shorter than those of MERS-CoV and SARS-CoV. These findings suggest the difficulty of controlling SARS-CoV-2 infection by antivirals. We further used the viral dynamics model to predict the efficacy of potential antiviral drugs that have different modes of action. The efficacy was measured by the reduction in the viral load area under the curve (AUC). Our results indicate that therapies that block de novo infection or virus production are likely to be effective if and only if initiated before the viral load peak (which appears 2–3 days after symptom onset), but therapies that promote cytotoxicity of infected cells are likely to have effects with less sensitivity to the timing of treatment initiation. Furthermore, combining a therapy that promotes cytotoxicity and one that blocks de novo infection or virus production synergistically reduces the AUC with early treatment. Our unique modeling approach provides insights into the pathogenesis of SARS-CoV-2 and may be useful for development of antiviral therapies. 
    more » « less
  5. ABSTRACT

    Within the contiguous USA, Florida is unique in having tropical and subtropical climates, a great abundance and diversity of mosquito vectors, and high rates of human travel. These factors contribute to the state being the national ground zero for exotic mosquito-borne diseases, as evidenced by local transmission of viruses spread by Aedes aegypti, including outbreaks of dengue in 2022 and Zika in 2016. Because of limited treatment options, integrated vector management is a key part of mitigating these arboviruses. Practical knowledge of when and where mosquito populations of interest exist is critical for surveillance and control efforts, and habitat predictions at various geographic scales typically rely on ecological niche modeling. However, most of these models, usually created in partnership with academic institutions, demand resources that otherwise may be too time-demanding or difficult for mosquito control programs to replicate and use effectively. Such resources may include intensive computational requirements, high spatiotemporal resolutions of data not regularly available, and/or expert knowledge of statistical analysis. Therefore, our study aims to partner with mosquito control agencies in generating operationally useful mosquito abundance models. Given the increasing threat of mosquito-borne disease transmission in Florida, our analytic approach targets recent Ae. aegypti abundance in the Tampa Bay area. We investigate explanatory variables that: 1) are publicly available, 2) require little to no preprocessing for use, and 3) are known factors associated with Ae. aegypti ecology. Out of our 4 final models, none required more than 5 out of the 36 predictors assessed (13.9%). Similar to previous literature, the strongest predictors were consistently 3- and 4-wk temperature and precipitation lags, followed closely by 1 of 2 environmental predictors: land use/land cover or normalized difference vegetation index. Surprisingly, 3 of our 4 final models included one or more socioeconomic or demographic predictors. In general, larger sample sizes of trap collections and/or citizen science observations should result in greater confidence in model predictions and validation. However, given disparities in trap collections across jurisdictions, individual county models rather than a multicounty conglomerate model would likely yield stronger model fits. Ultimately, we hope that the results of our assessment will enable more accurate and precise mosquito surveillance and control of Ae. aegypti in Florida and beyond.

     
    more » « less