Proteomic analysis of an Aedes albopictus cell line infected with Dengue serotypes 1 and 3 viruses (original) (raw)
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Proteomic Analysis of Host Responses in HepG2 Cells during Dengue Virus Infection
Journal of Proteome Research, 2007
Dengue virus infection remains a public health problem worldwide. However, its pathogenic mechanisms and pathophysiology are still poorly understood. We performed proteomic analysis to evaluate early host responses (as indicated by altered proteins) in human target cells during dengue virus infection. HepG2 cells were infected with dengue virus serotype 2 (DEN-2) at multiplicity of infection (MOI) of 0.1, 0.5, and 1.0. Quantitative analyses of DEN-2 infection and cell death at 12, 24, and 48 h postinfection showed that the MOI of 1.0 with 24 h postinfection duration was the optimal condition to evaluate early host responses, as this condition provided the high %Infection (∼80%), while %Cell death (∼20%) was comparable to that of the mock-control cells. Proteins derived from the mock-control and DEN-2-infected cells were resolved by 2-D PAGE (n ) 5 gels for each group) and visualized by SYPRO Ruby stain. Quantitative intensity analysis revealed 17 differentially expressed proteins, which were successfully identified by peptide mass fingerprinting. Most of these altered proteins were the key factors involved in transcription and translation processes. Further functional study on these altered proteins may lead to better understanding of the pathogenic mechanisms and host responses to dengue virus infection, and also to the identification of new therapeutic targets for dengue virus infection.
The role of the unfolded protein response in dengue virus pathogenesis
Cellular microbiology, 2017
Symptomatic dengue virus (DENV) infections range from mild fever to severe haemorrhagic disease and death. Host-viral interactions play a significant role in deciding the fate of the infection. The unfolded protein response (UPR) is a prosurvival cellular reaction induced in response to DENV-mediated endoplasmic reticulum stress. The UPR has complex interactions with the cellular autophagy machinery, apoptosis, and innate immunity. DENV has evolved to manipulate the UPR to facilitate its replication and to evade host immunity. Our knowledge of this intertwined network of events is continuously developing. A better understanding of the UPR mediated antiviral and proviral effects will shed light on dengue disease pathogenesis and may help development of anti-DENV therapeutics. This review summarizes the role of the UPR in viral replication, autophagy, and DENV-induced inflammation to describe how a host response contributes to DENV pathogenesis.
Quantitative proteomic analysis of Huh-7 cells infected with Dengue virus by label-free LC–MS
Journal of Proteomics, 2014
Dengue is an important and growing public health problem worldwide with an estimated 100 million new clinical cases annually. Currently, no licensed drug or vaccine is available. During natural infection in humans, liver cells constitute one of the main targets of dengue virus (DENV) replication. However, a clear understanding of dengue pathogenesis remains elusive. In order to gain a better reading of the cross talk between virus and host cell proteins, we used a proteomics approach to analyze the host response to DENV infection in a hepatic cell line Huh-7. Differences in proteome expression were assayed 24 h post-infection using label-free LC-MS. Quantitative analysis revealed 155 differentially expressed proteins, 64 of which were up-regulated and 91 down-regulated. These results reveal an important decrease in the expression of enzymes involved in the glycolytic pathway, citrate cycle, and pyruvate metabolism. This study provides large-scale quantitative information regarding protein expression in the early stages of infection that should be useful for better compression of the pathogenesis of dengue. Biological significance Dengue infection involves alterations in the homeostasis of the host cell. Defining the interactions between virus and cell proteins should provide a better understanding of how viruses propagate and cause disease. Here, we present for the first time the proteomic analysis of hepatocytes (Huh-7 cells) infected with DENV-2 by label-free LC-MS.
PLoS ONE, 2010
Background: Arthropod borne virus infections cause several emerging and resurgent infectious diseases. Among the diseases caused by arboviruses, dengue and chikungunya are responsible for a high rate of severe human diseases worldwide. The midgut of mosquitoes is the first barrier for pathogen transmission and is a target organ where arboviruses must replicate prior to infecting other organs. A proteomic approach was undertaken to characterize the key virus/vector interactions and host protein modifications that happen in the midgut for viral transmission to eventually take place.
The role of cell proteins in dengue virus infection
Despite 70 years of study, dengue disease continues to be a global health burden. Treatment is only supportive based on presenting symptoms. To date, there is no licensed prophylactic vaccine and no specific antiviral drugs available. The pathogenesis mechanisms during dengue virus infections remain poorly understood, and the complete picture on risk factors for developing severe clinical illness is still unknown. Viruses as obligate intracellular parasites depend on the host cell machinery for replication. As a result of a co-evolution process for million years, viruses have developed sophisticated strategies to hijack and use cellular factors for entry, replication and propagation, alternate host transmission and to combat host cell defenses. This review focuses on recent reports about cellular proteins involved along the dengue virus replication cycle, in prime cellular targets during the infection of both humans and mosquito hosts and also on the proteomics and other approaches that are being used to reveal the entire orchestration and most significant processes altered during infection. Identification of the key host cell factors involve in these processes will provide a better understanding of how viruses replicate and cause disease, and how to develop more effective therapeutic interventions.
Exploitation of cellular pathways by Dengue virus
Dengue virus (DENV) is the causative agent of the most prevalent arthropod-borne viral disease, thus representing a significant global health burden. Because of its limited coding capacity, DENV exploits components and pathways of the host cell to assure productive replication. In the past few years, important insights into this intimate interaction between DENV and the host cell have been gained. These include the identification of the ER-associated degradation pathway, autophagy, the unfolded protein response or lipid droplets that all play a crucial role for efficient DENV replication. In addition, strategies used by the virus to combat innate antiviral responses have been unraveled. Improving our understanding of the DENV-host cell relation will facilitate our attempts to develop efficient antiviral strategies.
BioMed Research International, 2013
The main vector of dengue in America is the mosquitoAedes aegypti, which is infected by dengue virus (DENV) through receptors of midgut epithelial cells. The envelope protein (E) of dengue virus binds to receptors present on the host cells through its domain III that has been primarily recognized to bind cell receptors. In order to identify potential receptors, proteins from mosquito midgut tissue and C6/36 cells were purified by affinity using columns with the recombinant E protein domain III (rE-DIII) or DENV particles bound covalently to Sepharose 4B to compare and evaluate their performance to bind proteins including putative receptors from female mosquitoes ofAe. aegypti. To determine their identity mass spectrometric analysis of purified proteins separated by polyacrylamide gel electrophoresis was performed. Our results indicate that both viral particles and rE-DIII bound proteins with the same apparent molecular weights of 57 and 67 kDa. In addition, viral particles bound hig...
Protein synthesized by dengue infected Aedes aegypti and Aedes albopictus
Tropical biomedicine, 2005
The main objective of this study was to compare protein profiles of whole mosquitoes of Malaysian Aedes aegypti and Aedes albopictus after infection with virus and to investigate whether dengue virus would induce protein secretion in Ae. aegypti and Ae. albopictus. Using SDS -PAGE, it was shown that in uninfected Ae. aegypti and Ae. albopictus, the protein bands were within the range of 14 - 80 kDa with most of the bands overlapping for the two species. Comparison of the protein profile of infected and uninfected Ae. aegypti and Ae. albopictus showed five distinct molecular weight grouping at 73 - 76 kDa (Group 1), 44 - 50 kDa (Group 2), 28 - 31 kDa (Group 3), 20 - 25 kDa (Group 4) and 14 - 17 kDa (Group 5). Predominant bands for both species (infected and uninfected) were between 21 - 25 kDa and 44 - 50 kDa. Protein bands having a molecular weight of 70 kDa were only present in infected Ae. albopictus and those bands having molecular weight of 21 kDa were observed only in infected ...
Monitoring Mitochondrial Function in Aedes albopictus C6/36 Cell Line during Dengue Virus Infection
Insects
Aedes aegypti and Aedes albopictus mosquitoes are responsible for dengue virus (DENV) transmission in tropical and subtropical areas worldwide, where an estimated 3 billion people live at risk of DENV exposure. DENV-infected individuals show symptoms ranging from sub-clinical or mild to hemorrhagic fever. Infected mosquitoes do not show detectable signs of disease, even though the virus maintains a lifelong persistent infection. The interactions between viruses and host mitochondria are crucial for virus replication and pathogenicity. DENV infection in vertebrate cells modulates mitochondrial function and dynamics to facilitate viral proliferation. Here, we describe that DENV also regulates mitochondrial function and morphology in infected C6/36 mosquito cells (derived from Aedes albopictus). Our results showed that DENV infection increased ROS (reactive oxygen species) production, modulated mitochondrial transmembrane potential and induced changes in mitochondrial respiration. Furt...
Proteomic Profile During Embryonic Development of Dengue Vector Aedes albopictus Mosquito
Biosciences Biotechnology Research Asia, 2014
Despite the potential impact of moisture on embryonation and egg eclosion of Aedes mosquitoes, little is known about its effect on protein synthesis during critical embryonic development as well as in the proteomic profiles. Thus, we quantify the protein concentration and proteomic profile during embryonic development of Ae. albopictus from far early of egg laying to egg eclosion in contact with sufficient moisture. It was observed that the concentration of protein started to decrease from the early hours (6th h) with progressing of embryonic development. There were more or less 13 bands observed in Coomassie blue staining of different embryonic stages within the range of 58 kDa and ~7 kDa by using 12% separating gel in 1D SDS-PAGE. Among them highly expressed bands on the position of 11-13 of lower molecular weight at around 7 kDa were found in all treatments. They may have controlling effects on egg hatching. Identification of these specific proteins can give an insight direction of effective vector control way.