RNA Interference Mediated Inhibition of Dengue Virus Multiplication and Entry in HepG2 Cells

Background: Dengue virus-host cell interaction initiates when the virus binds to the attachment receptors followed by endocytic internalization of the virus particle. Successful entry into the cell is necessary for infection initiation. Currently, there is no protective vaccine or antiviral treatment for dengue infection. Targeting the viral entry pathway has become an attractive therapeutic strategy to block infection. This study aimed to investigate the effect of silencing the GRP78 and clathrin-mediated endocytosis on dengue virus entry and multiplication into HepG2 cells. Methodology/Principal Findings: HepG2 cells were transfected using specific siRNAs to silence the cellular surface receptor (GRP78) and clathrin-mediated endocytosis pathway. Gene expression analysis showed a marked down-regulation of the targeted genes (87.2%, 90.3%, and 87.8 % for GRP78, CLTC, and DNM2 respectively) in transfected HepG2 cells when measured by RT-qPCR. Intracellular and extracellular viral RNA loads were quantified by RT-qPCR to investigate the effect of silencing the attachment receptor and clathrin-mediated endocytosis on dengue virus entry. Silenced cells showed a significant reduction of intracellular (92.4%) and extracellular viral RNA load (71.4%) compared to non-silenced cells. Flow cytometry analysis showed a marked reduction of infected cells (89.7%) in silenced HepG2 cells compared to non-silenced cells. Furthermore, the ability to generate infectious virions using the plaque assay was reduced 1.07 log in silenced HepG2 cells

Inhibition of Dengue Virus Entry and Multiplication into Monocytes Using RNA Interference

Prevention and treatment of dengue infection remain a serious global public health priority. Extensive efforts are required toward the development of vaccines and discovery of potential therapeutic compounds against the dengue viruses. Dengue virus entry is a critical step for virus reproduction and establishes the infection. Hence, the blockade of dengue virus entry into the host cell is an interesting antiviral strategy as it represents a barrier to suppress the onset of infection. This study was achieved by using RNA interference to silence the cellular receptor, and the clathrin mediated endocytosis that enhances the entry of dengue virus in monocytes. Results showed a marked reduction of infected monocytes by flow cytometry. In addition, both intracellular and extracellular viral RNA load was shown to be reduced in treated monocytes when compared to untreated monocytes. Based on these findings, this study concludes that this therapeutic strategy of blocking the virus replication at the first stage of multiplication might serve as a hopeful drug to mitigate the dengue symptoms, and reduction the disease severity

A household-based survey of knowledge, attitudes and practices towards dengue fever among local urban communities in Taiz Governorate, Yemen

BACKGROUND: Yemen has witnessed several dengue fever outbreaks coincident with the social unrest and war in the country. The aim of the present study was to describe the knowledge, attitudes and practices (KAPs) of at-risk urban populations residing in Taiz, southwest of Yemen. In addition, factors possibly associated with poor preventive practices were investigated. METHODS: A household-based, cross-sectional survey was conducted in three urban districts encompassing 383 households. Data on the socio-demographic characteristics and KAPs of the participating household heads were collected using a pre-designed, structured questionnaire. The association of socio-demographic characteristics, knowledge and attitudes of the population with poor preventive practices against dengue fever was then analyzed using logistic regression. RESULTS: More than 90.0 % of respondent household heads had correct knowledge about fever, headache and joint pain as common signs and symptoms of dengue fever. Moreover, muscular pain and bleeding were perceived by more than 80.0 % of the respondents as being associated with dengue fever; however, only 65.0 % of the respondents reported skin rash as a sign of dengue fever. More than 95.0 % of respondents agreed about the seriousness and possible transmission of dengue fever; however, negative attitudes regarding the facts of being at risk of the disease and that the infection is preventable were expressed by 15.0 % of respondents. Despite the good level of knowledge and attitudes of the respondent population, poor preventive practices were common. Bivariate analysis identified poor knowledge of dengue signs and symptoms (OR = 2.1, 95 % CI = 1.24–3.68; P = 0.005) and its vector (OR = 2.1, 95 % CI = 1.14–3.84; P = 0.016) as factors significantly associated with poor preventive practices. However, multivariable analysis showed that poor knowledge of the vector is an independent predictor of poor preventive practices of the population (adjusted OR = 2.1, 95 % CI = 1.14–3.84; P = 0.018). CONCLUSION: The majority of people in urban communities of Taiz have a clear understanding of most signs/symptoms of dengue fever as well as positive attitudes towards the seriousness and possible transmissibility of dengue fever. However, negative attitudes regarding their perception of the risk and possible prevention of the infection are prevailing among a small proportion of the population and need to be targeted by educational campaigns. It appears that the good level of the population knowledge of the signs/symptoms of dengue fever and the factors contributing to the spread and control of its vectors did not translate into good practices

Tackling dengue fever: Current status and challenges

International audienceAccording to recent statistics, 96 million apparent dengue infections were estimated worldwide in 2010. This figure is by far greater than the WHO prediction which indicates the rapid spread of this disease posing a growing threat to the economy and a major challenge to clinicians and health care services across the globe particularly in the affected areas. This article aims at bringing to light the current epidemiological and clinical status of the dengue fever. The relationship between genetic mutations, single nucleotide polymorphism (SNP) and the pathophysiology of disease progression will be put into perspective. It will also highlight the recent advances in dengue vaccine development. Thus far, a significant progress has been made in unraveling the risk factors and understanding the molecularpathogenesis associated with the disease. However, further insights in molecular features of the disease and the development of animal models will enormously help improving the therapeutic interventions and potentially contribute to finding new preventive measures for population at risk