Mosquito-borne Dengue Virus Research Articles

Dengue virus preferentially uses human and mosquito non-optimal codons

Codon optimality refers to the effect that codon composition has on messenger RNA (mRNA) stability and translation level and implies that synonymous codons are not silent from a regulatory point of view. Here, we investigated the adaptation of virus genomes to the host optimality code using mosquito-borne dengue virus (DENV) as a model. We demonstrated that codon optimality exists in mosquito cells and showed that DENV preferentially uses nonoptimal (destabilizing) codons and avoids codons that are defined as optimal (stabilizing) in either human or mosquito cells. Human genes enriched in the codons preferentially and frequently used by DENV are upregulated during infection, and so is the tRNA decoding the nonoptimal and DENV preferentially used codon for arginine. We found that adaptation during single-host passaging in human or mosquito cells results in the selection of synonymous mutations towards DENV’s preferred nonoptimal codons that increase virus fitness. Finally, our analyses revealed that hundreds of viruses preferentially use nonoptimal codons, with those infecting a single host displaying an even stronger bias, suggesting that host–pathogen interaction shapes virus-synonymous codon choice.

Emergence of Indigenous Dengue Fever, Niger, October 2023.

Dengue fever is a growing worldwide public health concern. In mid-October 2023, multiple cases of uncommon febrile illness were reported among patients in Niamey, Niger. Fifteen samples were tested by using molecular methods, from which 7 (46.66%) were confirmed positive for mosquitoborne dengue virus belonging to serotypes 1 and 3.

Bangladesh records persistently increased number of dengue deaths in recent years: Dissecting the shortcomings and means to resolve

ObjectivesDengue, a life-threatening disease caused by the mosquito-borne dengue virus, has become a severe problem in recent years in Bangladesh, a South Asian country. In this study, we have critically analyzed the factors contributing to the escalation of the dengue burden in recent years in Bangladesh and discussed the strategies for effective control. DesignThis retrospective observational study analyzed the dengue data collected from the five most dengue-affected countries (Bangladesh, Brazil, Mexico, Peru, and the Philippines) from 2019 to 2023. ResultsAn increased number of dengue-related deaths has been recorded in Bangladesh since 2021. The year 2023 has witnessed a record high dengue-related deaths in Bangladesh, with cumulative deaths for the year surpassing all totals of the previous 23 years (2000-2022: 853 deaths vs 2023: 1705 deaths). Comparing the epidemiologic data of major dengue-endemic countries over the last 5 years, Bangladesh recorded higher dengue fatality rates consecutively for 3 years. Besides the environmental and viral host factors, which are also applicable to many other dengue-endemic countries, there is concern about the failures and mismanagement of authorities to manage dengue patients properly. ConclusionThis study provided evidence that Bangladesh recorded higher dengue fatality rates in recent years. By implementing multi-pronged proactive approaches that can ensure proper prevention programs and appropriate patient management, Bangladesh or similar other countries can significantly reduce the current dengue burden and the associated deaths.

A Coupled Statistical and Deterministic Model for Forecasting Climate-Driven Dengue Incidence in Selangor, Malaysia.

The mosquito-borne dengue virus remains a major public health concern in Malaysia. Despite various control efforts and measures introduced by the Malaysian Government to combat dengue, the increasing trend of dengue cases persists and shows no sign of decreasing. Currently, early detection and vector control are the main methods employed to curb dengue outbreaks. In this study, a coupled model consisting of the statistical ARIMAX model and the deterministic SI-SIR model was developed and validated using the weekly reported dengue data from year 2014 to 2019 for Selangor, Malaysia. Previous studies have shown that climate variables, especially temperature, humidity, and precipitation, were able to influence dengue incidence and transmission dynamics through their effect on the vector. In this coupled model, climate is linked to dengue disease through mosquito biting rate, allowing real-time forecast of dengue cases using climate variables, namely temperature, rainfall and humidity. For the period chosen for model validation, the coupled model can forecast 1-2weeks in advance with an average error of less than 6%, three weeks in advance with an average error of 7.06% and four weeks in advance with an average error of 8.01%. Further model simulation analysis suggests that the coupled model generally provides better forecast than the stand-alone ARIMAX model, especially at the onset of the outbreak. Moreover, the coupled model is more robust in the sense that it can be further adapted for investigating the effectiveness of various dengue mitigation measures subject to the changing climate.

Trade-offs shaping transmission of sylvatic dengue and Zika viruses in monkey hosts

Mosquito-borne dengue (DENV) and Zika (ZIKV) viruses originated in Old World sylvatic (forest) cycles involving monkeys and canopy-living Aedes mosquitoes. Both viruses spilled over into human transmission and were translocated to the Americas, opening a path for spillback into Neotropical sylvatic cycles. Studies of the trade-offs that shape within-host dynamics and transmission of these viruses are lacking, hampering efforts to predict spillover and spillback. We infected a native, Asian host species (cynomolgus macaque) and a novel, American host species (squirrel monkey) with sylvatic strains of DENV-2 or ZIKV via mosquito bite. We then monitored aspects of viral replication (viremia), innate and adaptive immune response (natural killer (NK) cells and neutralizing antibodies, respectively), and transmission to mosquitoes. In both hosts, ZIKV reached high titers that translated into high transmission to mosquitoes; in contrast DENV-2 replicated to low levels and, unexpectedly, transmission occurred only when serum viremia was below or near the limit of detection. Our data reveal evidence of an immunologically-mediated trade-off between duration and magnitude of virus replication, as higher peak ZIKV titers are associated with shorter durations of viremia, and higher NK cell levels are associated with lower peak ZIKV titers and lower anti-DENV-2 antibody levels. Furthermore, patterns of transmission of each virus from a Neotropical monkey suggest that ZIKV has greater potential than DENV-2 to establish a sylvatic transmission cycle in the Americas.

AN IN-DEPTH ANALYSIS OF DENGUE MOSQUITO DYNAMICS IN LAHORE: UNRAVELING THE IMPACT OF DENGUE OUTBREAKS AND ENVIRONMENTAL FACTORS

Dengue fever, caused by the Aedes mosquito-borne dengue virus, is still a major public health concern in many parts of the world, including Lahore, Pakistan. Lahore region is regarded as a dengue fever hotspot due to the large number of cases reported each year. The purpose of this research is to give an in-depth analysis of dengue mosquito dynamics in Lahore, with a particular emphasis on determining the influence of dengue epidemics and environmental factors. The current study was based on survey comprising 240 individuals from Lahore. In the study findings, 82.08% were positive cases and 17.91% were negative cases. The majority of respondents, were aware of the symptoms of dengue infection (62.33%), its transmission (90.83%), the cause of dengue (61%), safety precautions (73.33%), and the organ infected by the dengue virus (66.25). However, the majority of people are still clueless. 70% of participants indicate that television is their primary information source. Dengue fever cases have been found to have a seasonal pattern, peaking during the monsoon season (45.41%) when mosquito breeding conditions are perfect. The study's findings revealed that Aedes mosquitos, particularly Aedes aegypti, were widespread and identified as the primary vectors (61.66%) of dengue transmission in Lahore. It was revealed that temperature (49.58%), and rainfall all affected the number of Aedes mosquitoes. Overall, 81.6% were satisfied with Governmental efforts against the spread of dengue fever. Improved public knowledge is necessary to reduce dengue virus incidence. The findings of this study can assist public health professionals and legislators in developing targeted measures to mitigate the consequences of dengue epidemics and protect the public from this terrible infection.

Larvicidal activity of CuO and ZnO nanoparticles against Aedes aegypti and Anopheles stephensi mosquito vectors-a greener approach by Phaseolus vulgaris L. aqueous extract as bio-reductant

In order to eradicate mosquito-borne diseaseslikemalaria,chikungunya, dengue,and Zika virus which are mostly found in temperate zones, synthetic pesticides are widely used as a form of fumigation. This has resulted in a rise in mosquito-borne disease transmission dynamics and a substantial rise in atmospheric pollution. In light of this, the current work used an aqueous extract of Phaseolus vulgaris to formulate bioencapsulated CuO and ZnO NPs to combat mosquito vectors. The synthesized CuO and ZnO NPs were characterized using UV–visible, XRD, FT-IR, FE-SEM & EDX, AFM, and DLS techniques. According to analytical characterization, the CuO and ZnO NPs had monoclinic and hexagonal crystallites with average sizesof 13.7 nm and 185 nm, respectively. FE-SEM analysis confirms the spherical and flower-like morphology of CuO and ZnO NPs and their respective elemental composition was identified by EDX studies. The average particle size of CuO and ZnO NPs was determined to be 22.6 nm and 662 nm. CuO and ZnO NPs possessed zeta potential values of −23.2 mV and −29.7 mV respectively. The larvae of the malaria vector Anopheles stephensi and the dengue vector Aedes aegypti were subjected to compare the larvicidal bioassays in a range of concentrations of P. vulgaris aqueous extract with methanol extracts, and CuO NPs with ZnO NPs (6.25, 12.5, 25, 50, 100 µg/mL). The results showed the larvicidal activity of the aqueous extract is superior to the methanol extract and ZnO NPs are superior to the CuO NPs. This study further revealed that Aedes aegypti vulnerable than Anopheles stephensi to a given concentration of crude aqueous, methanol extracts of P. vulgaris and to CuO and ZnO NPs. The overall findings of this research demonstrated the feasibility of using these biosynthesized nanoparticles as highly effective, eco-friendly natural mosquito control pesticides.

Spatial analysis in epidemiological studies of dengue: A systematic review and meta-analysis

Background: Dengue is a serious tropical disease caused by the mosquito-borne dengue virus (DENV). Over the last 50 years, the incidence of dengue fever (DF) and dengue fever and dengue hemorrhagic fever (DHF) worldwide has increased by 50-fold and 2.5 billion people currently live in dengue-endemic regions. This study reports a systematic review and meta-analysis of different types of applications offered by spatial analysis in epidemiological studies of dengue. Materials and methods: A comprehensive literature search of Web of Science, SCOPUS, Google Scholar, and PubMed were conducted up to December 2023. A total of three sub-sections, namely spatial analysis of points, spatial analysis of area data, and software programs used for spatial analysis of dengue cases, will be presented in this review. Results: It was found that spatial analysis can effectively support the study of dengue fever and dengue hemorrhagic fever such as to investigate spatial patterns, hotspot, spatial clustering, the spread, transmission of dengue fever and dengue hemorrhagic fever and it’s affecting factors. Conclusion: applications of spatial analysis can provide important information and contribute to development of effective measurements to control and prevent the dengue fever and dengue hemorrhagic fever transmission. Finding in this study provide an insight into how to use different techniques in the study of dengue fever and dengue hemorrhagic fever.

Effect of previous heterologous flavivirus vaccinations on human antibody responses in tick-borne encephalitis and dengue virus infections.

Arthropod-borne flaviviruses include a number of medically relevant human pathogens such as the mosquito-borne dengue (DEN), Zika, and yellow fever (YF) viruses as well as tick-borne encephalitis virus (TBEV). All flaviviruses are antigenically related and anamnestic responses due to prior immunity can modulate antibody specificities in subsequent infections or vaccinations. In our study, we analyzed the induction of broadly flavivirus cross-reactive antibodies in tick-borne encephalitis (TBE) and DEN patients without or with prior flavivirus exposure through TBE and/or YF vaccination, and determined the contribution of these antibodies to TBE and dengue virus (DENV) neutralization. In addition, we investigated the formation of cross-reactive antibodies in TBE-vaccination breakthroughs (VBTs). A TBEV infection without prior YF or TBE vaccination induced predominantly type-specific antibodies. In contrast, high levels of broadly cross-reactive antibodies were found in samples from TBE patients prevaccinated against YF as well as in DEN patients prevaccinated against TBE and/or YF. While these cross-reactive antibodies did not neutralize TBEV, they were effective in neutralizing DENV. This discrepancy points to structural differences between the two viruses and indicates that broadly cross-reactive epitopes are less accessible in TBEV than in DENV. In TBE VBT infections, type-specific antibodies dominated the antibody response, thus revealing no difference from that of unvaccinated TBE patients. Our results emphasize significant differences in the structural properties of different flaviviruses that have an impact on the induction of broadly cross-reactive antibodies and their functional activities in virus neutralization.

Exposure to ultraviolet-B radiation increases the susceptibility of mosquitoes to infection with dengue virus.

By 2100, greenhouse gases are predicted to reduce ozone and cloud cover over the tropics causing increased exposure of organisms to harmful ultraviolet-B radiation (UVBR). UVBR damages DNA and is an important modulator of immune function and disease susceptibility in humans and other vertebrates. The effect of UVBR on invertebrate immune function is largely unknown, but UVBR together with ultraviolet-A radiation impairs an insect immune response that utilizes melanin, a pigment that also protects against UVBR-induced DNA damage. If UVBR weakens insect immunity, then it may make insect disease vectors more susceptible to infection with pathogens of socioeconomic and public health importance. In the tropics, where UVBR is predicted to increase, the mosquito-borne dengue virus (DENV), is prevalent and a growing threat to humans. We therefore examined the effect of UVBR on the mosquito Aedes aegypti, the primary vector for DENV, to better understand the potential implications of increased tropical UVBR for mosquito-borne disease risk. We found that exposure to a UVBR dose that caused significant larval mortality approximately doubled the probability that surviving females would become infected with DENV, despite this UVBR dose having no effect on the expression of an effector gene involved in antiviral immunity. We also found that females exposed to a lower UVBR dose were more likely to have low fecundity even though this UVBR dose had no effect on larval size or activity, pupal cuticular melanin content, or adult mass, metabolic rate, or flight capacity. We conclude that future increases in tropical UVBR associated with anthropogenic global change may have the benefit of reducing mosquito-borne disease risk for humans by reducing mosquito fitness, but this benefit may be eroded if it also makes mosquitoes more likely to be infected with deadly pathogens.

Dengue virus serotypic replacement of NS3 protease or helicase domain causes chimeric viral attenuation but can be recovered by a compensated mutation at helicase domain or NS2B, respectively.

Mosquito-borne dengue viruses (DENVs) have evolved to four serotypes with 69%-78% amino acid identities, resulting in incomplete immunity, where one serotype's infection does not cross-protect against secondary infections by other serotypes. Despite the amino acid differences, structural and nonstructural (NS) proteins among serotypes play similar functions. NS3 is an enzyme complex: NS3 has N-terminal protease (PRO) and C-terminal helicase (HEL) activities in addition to 5' RNA triphosphatase (5'RTP), which is involved in the RNA capping process. In this study, the effects of NS3 replacements among serotypes were tested. The replacement of NS3 full-length (FULL), PRO or HEL region suppressed viral replication in BHK-21 mammalian cells, while the single compensatory mutation improved the viral replications; P364S mutation in HEL revived PRO (DENV3)-replaced DENV1, while S68T alteration in NS2B recovered HEL (DENV1)-replaced DENV2. The results suggest that the interactions between PRO and HEL as well as HEL and NS2B are required for replication competence. Lower-frequency mutations also appeared at various locations in viral proteins, although after infecting C6/36 mosquito cells, the mutations' frequencies changed, and/or new mutations appeared. In contrast, the inter-domain region (INT, 12 amino acids)-replaced chimera quickly replicated without mutation in BHK-21 cells, although extended cell culture accumulated various mutations. These results suggest that NS3 variously interacts with DENV proteins, in which the chimeric NS3 domain replacements induced amino acid mutations, irrespective of replication efficiency. However, the viral sequences are further adjusted for replication efficiency, to fit in both mammalian cells and mosquito cells. IMPORTANCE Enzyme activities for replicating DENV 5' cap positive (+) sense RNA have been shown to reside in NS3 and NS5. However, it remains unknown how these enzymes coordinately synthesize negative (-) sense RNA, from which abundant 5' cap (+) sense RNA is produced. We previously revealed that NS5 dimerization and NS5 methyltransferase(MT)-NS3HEL interaction are important for DENV replication. Here, we found that replication incompetence due to NS3PRO or HEL replacement was compensated by a mutation at HEL or NS2B, respectively, suggesting that the interactions among NS2B, NS3PRO, and HEL are critical for DENV replication.

Co-infection of dengue and Zika viruses mutually enhances viral replication in the mosquito Aedes aegypti

BackgroundThe mosquito Aedes aegypti transmits two of the most serious mosquito-borne viruses, dengue virus (DENV) and Zika virus (ZIKV), which results in significant human morbidity and mortality worldwide. The quickly shifting landscapes of DENV and ZIKV endemicity worldwide raise concerns that their co-circulation through the Ae. aegypti mosquito vector could greatly exacerbate the disease burden in humans. Recent reports have indicated an increase in the number of co-infection cases in expanding co-endemic regions; however, the impact of co-infection on viral infection and the detailed molecular mechanisms remain to be defined.MethodsC6/36 (Aedes albopictus) cells were cultured in Dulbecco's modified Eagle medium/Mitsuhashi and Maramorosch Insect Medium (DMEM/MM) (1:1) containing 2% heat-inactivated fetal bovine serum and 1× penicillin/streptomycin solution. For virus propagation, the cells were infected with either DENV serotype 2 (DENV2) strain 16681 or ZIKV isolate Thailand/1610acTw (MF692778.1). Mosquitoes (Ae. aegypti UGAL [University of Georgia Laboratory]/Rockefeller strain) were orally infected with DENV2 and ZIKV through infectious blood-feeding.ResultsWe first examined viral replication activity in cells infected simultaneously, or sequentially, with DENV and ZIKV, and found interspecies binding of viral genomic transcripts to the non-structural protein 5 (NS5). When we challenged Ae. aegypti mosquitos with both DENV2 and ZIKV sequentially to probe similar interactions, virus production and vector susceptibility to infection were significantly enhanced.ConclusionsOur results suggest that DENV2 and ZIKV simultaneously establishing infection in the Ae. aegypti mosquito vector may augment one another during replication. The data also implicate the homologous NS5 protein as a key intersection between the flaviviruses in co-infection, highlighting it as a potential target for vector control.

DDPM: A Dengue Disease Prediction and Diagnosis Model Using Sentiment Analysis and Machine Learning Algorithms

The aedes mosquito-borne dengue viruses cause dengue fever, an arboviral disease (DENVs). In 2019, the World Health Organization forecasts a yearly occurrence of infections from 100 million to 400 million, the maximum number of dengue cases ever testified worldwide, prompting WHO to label the virus one of the world's top ten public health risks. Dengue hemorrhagic fever can progress into dengue shock syndrome, which can be fatal. Dengue hemorrhagic fever can also advance into dengue shock syndrome. To provide accessible and timely supportive care and therapy, it is necessary to have indispensable practical instruments that accurately differentiate Dengue and its subcategories in the early stages of illness development. Dengue fever can be predicted in advance, saving one's life by warning them to seek proper diagnosis and treatment. Predicting infectious diseases such as dengue is difficult, and most forecast systems are still in their primary stages. In developing dengue predictive models, data from microarrays and RNA-Seq have been used significantly. Bayesian inferences and support vector machine algorithms are two examples of statistical methods that can mine opinions and analyze sentiment from text. In general, these methods are not very strong semantically, and they only work effectively when the text passage inputs are at the level of the page or the paragraph; they are poor miners of sentiment at the level of the sentence or the phrase. In this research, we propose to construct a machine learning method to forecast dengue fever.

Prediction of dengue incidents using hospitalized patients, metrological and socio-economic data in Bangladesh: A machine learning approach.

Dengue fever is a severe disease spread by Aedes mosquito-borne dengue viruses (DENVs) in tropical areas such as Bangladesh. Since its breakout in the 1960s, dengue fever has been endemic in Bangladesh, with the highest concentration of infections in the capital, Dhaka. This study aims to develop a machine learning model that can use relevant information about the factors that cause Dengue outbreaks within a geographic region. To predict dengue cases in 11 different districts of Bangladesh, we created a DengueBD dataset and employed two machine learning algorithms, Multiple Linear Regression (MLR) and Support Vector Regression (SVR). This research also explores the correlation among environmental factors like temperature, rainfall, and humidity with the rise and decline trend of Dengue cases in different cities of Bangladesh. The entire dataset was divided into an 80:20 ratio, with 80 percent used for training and 20% used for testing. The research findings imply that, for both the MLR with 67% accuracy along with Mean Absolute Error (MAE) of 4.57 and SVR models with 75% accuracy along with Mean Absolute Error (MAE) of 4.95, the number of dengue cases reduces throughout the winter season in the country and increases mainly during the rainy season in the next ten months, from August 2021 to May 2022. Importantly, Dhaka, Bangladesh’s capital, will see the maximum number of dengue patients during this period. Overall, the results of this data-driven analysis show that machine learning algorithms have enormous potential for predicting dengue epidemics.

Cranberry Pomace Extract Exerts Antiviral Activity against Zika and Dengue Virus at Safe Doses for Adult Zebrafish.

Mosquito-borne dengue virus (DENV) and zika virus (ZIKV) infections constitute a global health emergency. Antivirals directly targeting the virus infectious cycle are still needed to prevent dengue hemorrhagic fever and congenital zika syndrome. In the present study, we demonstrated that Cranberry Pomace (CP) extract, a polyphenol-rich agrifood byproduct recovered following cranberry juice extraction, blocks DENV and ZIKV infection in human Huh7.5 and A549 cell lines, respectively, in non-cytotoxic concentrations. Our virological assays identified CP extract as a potential inhibitor of virus entry into the host-cell by acting directly on viral particles, thus preventing their attachment to the cell surface. At effective antiviral doses, CP extract proved safe and tolerable in a zebrafish model. In conclusion, polyphenol-rich agrifood byproducts such as berry extracts are a promising source of safe and naturally derived nutraceutical antivirals that target medically important pathogens.

Vitamin D-induced LL-37 modulates innate immune responses of human primary macrophages during DENV-2 infection.

Epidemics of dengue, an acute and potentially severe disease caused by mosquito-borne dengue virus (DENV), pose a major challenge to clinicians and health care services across the sub(tropics). Severe disease onset is associated with a dysregulated inflammatory response to the virus, and there are currently no drugs to alleviate disease symptoms. LL-37 is a potent antimicrobial peptide with a wide range of immunoregulatory properties. In this study, we assessed the effect of LL-37 on DENV-2-induced responses in human monocyte-derived macrophages (MDMs). We show that simultaneous exposure of exogenous LL-37 and DENV-2 resulted in reduced replication of the virus in MDMs, while the addition of LL-37 postexposure to DENV-2 did not. Interestingly, the latter condition reduced the production of IL-6 and increased the expression of genes involved in virus sensing and antiviral response. Finally, we demonstrate that low endogenous levels and limited production of LL-37 in MDMs in response to DENV-2 infection can be increased by differentiating MDMs in the presence of Vitamin D (VitD3). Taken together, this study demonstrates that in addition to its antimicrobial properties, LL-37 has immunomodulatory properties in the curse of DENV infection and its production can be increased by VitD3.

Molecular Organisation of Tick-Borne Encephalitis Virus.

Tick-borne encephalitis virus (TBEV) is a pathogenic, enveloped, positive-stranded RNA virus in the family Flaviviridae. Structural studies of flavivirus virions have primarily focused on mosquito-borne species, with only one cryo-electron microscopy (cryo-EM) structure of a tick-borne species published. Here, we present a 3.3 Å cryo-EM structure of the TBEV virion of the Kuutsalo-14 isolate, confirming the overall organisation of the virus. We observe conformational switching of the peripheral and transmembrane helices of M protein, which can explain the quasi-equivalent packing of the viral proteins and highlights their importance in stabilising membrane protein arrangement in the virion. The residues responsible for M protein interactions are highly conserved in TBEV but not in the structurally studied Hypr strain, nor in mosquito-borne flaviviruses. These interactions may compensate for the lower number of hydrogen bonds between E proteins in TBEV compared to the mosquito-borne flaviviruses. The structure reveals two lipids bound in the E protein which are important for virus assembly. The lipid pockets are comparable to those recently described in mosquito-borne Zika, Spondweni, Dengue, and Usutu viruses. Our results thus advance the understanding of tick-borne flavivirus architecture and virion-stabilising interactions.

Tracing and tracking the emergence, epidemiology and dispersal of dengue virus to Africa during the 20th century

The four mosquito-borne dengue virus serotypes (DENV1–DENV4) cause a high burden of disease throughout the tropical and sub-tropical regions of the world. Nevertheless, their precise epidemiological history in Africa, including when and where they originated and were distributed during the 20th century, remains unclear stressing the need for One Health focused research. Accordingly, we conducted a time-scaled molecular epidemiological reconstruction using publicly available and newly sequenced dengue virus genomes of African origin representing all four serotypes to deduce the most likely temporal and spatial transmission routes of each DENV serotype from their ancestral regions to, within and from Africa.Our analyses suggest that during the 20th century, serotypes DENV1–DENV3 were introduced to Africa from South East Asia on multiple occasions. The earliest evidence recorded indicates introduction of DENV2 during the early-1940s and of DENV1 during the mid-1940s to Western Africa from South East Asia. The analysis also implies an early introduction of DENV4 during the mid-1940s to Western Africa, alongside DENV1, probably originating in South East Asia. Establishment of DENV3 in Africa appears to have occurred later in the 1960s, apparently originating from South East Asia. However, with the re-establishment of DENV in the Americas, following the cessation of the PAHO mosquito control programme during the mid-20th century, evidence of introductions of DENV1 and DENV2 from the Americas to Western Africa was also observed. The data also identify intra-regional circulation of DENV, but also inter-regional dispersal of all four serotypes within Africa, which has led to a high degree of geographical overlap among serotypes. It is also noteworthy that DENV from both Eastern and Western Africa, have been introduced into Central Africa but there is no support for the converse relationship. For serotypes DENV1–DENV3, we observed probable exports from within established African DENV clusters (≥2 sequences) primarily to Eastern and Southern Asia.Collectively, our findings support the view that all DENV serotypes, apart from DENV4, have been introduced on multiple occasions to Africa, primarily originating from South East Asia, and subsequently to neighbouring regions within Africa.

Regulation of innate immune responses in macrophages differentiated in the presence of vitamin D and infected with dengue virus 2.

A dysregulated or exacerbated inflammatory response is thought to be the key driver of the pathogenesis of severe disease caused by the mosquito-borne dengue virus (DENV). Compounds that restrict virus replication and modulate the inflammatory response could thus serve as promising therapeutics mitigating the disease pathogenesis. We and others have previously shown that macrophages, which are important cellular targets for DENV replication, differentiated in the presence of bioactive vitamin D (VitD3) are less permissive to viral replication, and produce lower levels of pro-inflammatory cytokines. Therefore, we here evaluated the extent and kinetics of innate immune responses of DENV-2 infected monocytes differentiated into macrophages in the presence (D3-MDMs) or absence of VitD3 (MDMs). We found that D3-MDMs expressed lower levels of RIG I, Toll-like receptor (TLR)3, and TLR7, as well as higher levels of SOCS-1 in response to DENV-2 infection. D3-MDMs produced lower levels of reactive oxygen species, related to a lower expression of TLR9. Moreover, although VitD3 treatment did not modulate either the expression of IFN-α or IFN-β, higher expression of protein kinase R (PKR) and 2'-5'-oligoadenylate synthetase 1 (OAS1) mRNA were found in D3-MDMs. Importantly, the observed effects were independent of reduced infection, highlighting the intrinsic differences between D3-MDMs and MDMs. Taken together, our results suggest that differentiation of MDMs in the presence of VitD3 modulates innate immunity in responses to DENV-2 infection.

Evaluation of VIDAS® Diagnostic Assay Prototypes Detecting Dengue Virus NS1 Antigen and Anti-Dengue Virus IgM and IgG Antibodies.

Dengue is a serious tropical disease caused by the mosquito-borne dengue virus (DENV). Performant, rapid, and easy-to-use assays are needed for the accurate diagnosis of acute DENV infection. We evaluated the performance of three prototype assays developed for the VIDAS® automated platform to detect dengue NS1 antigen and anti-dengue IgM and IgG antibodies. Positive and negative agreement with competitor enzyme-linked immunosorbent assays (ELISA) and rapid diagnostic tests (RDT) was evaluated in 91 Lao patients (57 adults, 34 children) with acute DENV infection. The VIDAS® NS1 assay showed the best overall agreement (95.6%) with the competitor NS1 ELISA. Both VIDAS® NS1 and NS1 ELISA assays also demonstrated high sensitivity relative to DENV RNA RT-PCR set as gold standard (85.7% and 83.9%, respectively). In contrast, NS1 RDT was less sensitive relative to DENV RNA RT-PCR (72.7%). The overall agreement of VIDAS® IgM and IgG assays with the competitor assays was moderate (72.5% for IgM ELISA, 76.9% for IgG ELISA, and 68.7% for IgM and IgG RDT). In most analyses, test agreements of the VIDAS® assays were comparable in adults and children. Altogether, the VIDAS® dengue prototypes performed very well and appear to be suitable for routine detection of dengue NS1 antigen and anti-dengue IgM/IgG antibodies.