Autor(es): Kang, Seokyoung; Shields, Alicia R.; Jupatanakul, Natapong; Dimopoulos, George

Resumo: Dengue virus host factors (DENV HFs) that are essential for the completion of the infection cycle in the mosquito vector and vertebrate host represent potent targets for transmission blocking. Here we investigated whether known mammalian DENV HF inhibitors could influence virus infection in the arthropod vector A. aegypti. We evaluated the potency of bafilomycin (BAF; inhibitor of vacuolar H+-ATPase (vATPase)), mycophenolic acid (MPA; inhibitor of inosine-5'-monophosphate dehydrogenase (IMPDH)), castanospermine (CAS; inhibitor of glucosidase), and deoxynojirimycin (DNJ; inhibitor of glucosidase) in blocking DENV infection of the mosquito midgut, using various treatment methods that included direct injection, ingestion by sugar feeding or blood feeding, and silencing of target genes by RNA interference (RNAi). Injection of BAF (5 mu M) and MPA (25 mu M) prior to feeding on virus-infected blood inhibited DENV titers in the midgut at 7 days post-infection by 56% and 60%, and in the salivary gland at 14 days post-infection by 90% and 83%, respectively, while treatment of mosquitoes with CAS or DNJ did not affect susceptibility to the virus. Ingestion of BAF and MPA through a sugar meal or together with an infectious blood meal also resulted in various degrees of virus inhibition. RNAi-mediated silencing of several vATPase subunit genes and the IMPDH gene resulted in a reduced DENV infection, thereby indicating that BAF- and MPA-mediated virus inhibition in adult mosquitoes most likely occurred through the inhibition of these DENV HFs. The route and timing of BAF and MPA administration was essential, and treatment after exposure to the virus diminished the antiviral effect of these compounds. Here we provide proof-of-principle that chemical inhibition or RNAi-mediated depletion of the DENV HFs vATPase and IMPDH can be used to suppress DENV infection of adult A. aegypti mosquitoes, which may translate to a reduction in DENV transmission. Arboviruses utilize homologous host factors of the mammalian and insect cellular machinery to complete the infection cycle. Studies in both mammalian and insect cell lines have shown that virus infection can be suppressed through inhibition of host factors by chemical compounds that therefore could be developed into transmission blocking agents. However, similar studies have not been conducted in adult mosquitoes. Here we investigated the effect of four chemical compounds (bafilomycin, mycophenolic acid, castanospermine, and deoxynojirimycin), known to inhibit the host factors vacuolar H+-ATPase (vATPase), inosine-5'-monophosphate dehydrogenase (IMPDH) and glucosidases, on dengue virus replication in adult mosquitoes. We found that bafilomycin and mycophenolic acid suppressed dengue virus replication in adult mosquito guts when they were injected prior to dengue virus infection; however, castanospermine and deoxynojirimycin did not. Ingestion of bafilomycin and mycophenolic acid also inhibited virus replication. We showed that the predicted target genes of bafilomycin and mycophenolic acid function as virus host factors in adult mosquitoes through RNAi-mediated gene silencing. Inhibition of vATPase also decreases mosquito longevity and fecundity, thereby further compromising vector capacity. Our study demonstrated that chemical compounds or double stranded RNAs (dsRNA) can be used to suppress virus infection through inhibition of host factors in adult mosquitoes, thereby rendering such approaches interesting for the development of novel transmission-blocking strategies.

Palavras-Chave: Human diseases; Replication; Inhibitors; Pest control; Hosts; Chemical compounds; Aquatic insects; Disease transmission; Public health; Feeding; Sugar; Mycophenolic acid; Double-stranded RNA; Vectors; Blood meals; Salivary gland; Infection; Longevity; Fecundity; Digestive tract; Insect cells; RNA-mediated interference; Midgut; H super(+)-transporting ATPase; Gene silencing; Dengue; Machinery; Arthropods; Ingestion; Insects; Dengue virus; Aedes aegypti; Arthropoda; Dengue virus type 2

Imprenta: Plos Neglected Tropical Diseases, v. 8, n. 8, 2014.

Descritores: Aedes aegypti - Cell ; Aedes aegypti - RNA ; Aedes aegypti - Infectious diseases ; Aedes aegypti - Molecular methods ; Aedes aegypti - Virus ; Aedes aegypti - Transmission ; Aedes aegypti - Dengue ; Aedes aegypti - Immunology ; Aedes aegypti - Public health

Data de publicação: 2014

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