Isolation of midgut escape mutants of two American genotype dengue 2 viruses from Aedes aegypti.

Autor(es): Khoo Cynthia C H; Doty Jeffrey B; Held Nicole L; Olson Ken E; Franz Alexander W E

Resumo: Several studies have shown that American genotype dengue 2 viruses (DENV2) have reduced viral fitness in the mosquito vector, Aedes aegypti, compared to other DENV2 genotypes. Diminished replication efficiency or inability to efficiently traverse membrane barriers encompassing organs such as the midgut or salivary glands are considered major factors negatively impacting viral fitness in the mosquito. We analyzed the vector competence of Ae. aegypti for two American DENV2 strains, QR94 and PR159 originating from Mexico and Puerto-Rico, respectively. Both strains infected mosquito midguts following acquisition of infectious bloodmeals. However, DENV2-QR94 and DENV2-PR159 poorly disseminated from the midgut at 7 or 14 days post-bloodmeal (pbm). We detected one virus isolate, EM33, among 31 DENV2-QR94 infected mosquitoes, and one isolate, EM41, among 121 DENV2-PR159 infected mosquitoes, generating high virus titers in mosquito carcasses at 7 days pbm. In oral challenge experiments, EM33 and EM41 showed midgut dissemination rates of 40-50%. Replication efficiency of EM41 in secondary mosquito tissue was similar to that of a dissemination-competent control strain, whereas the replication efficiency of EM33 was significantly lower than that of the control virus. The genome sequence of DENV2-QR94 encoded seven unique amino acids (aa), which were not found in 100 of the most closely related DENV2 strains. EM33 had one additional aa change, E202K, in the E protein. DENV2-PR159 encoded four unique aa residues, one of them E202K, whereas EM41 had two additional aa substitutions, Q77E in the E protein and E93D in NS3. Our results indicate that the midgut of Ae. aegypti acts as a selective sieve for DENV2 in which genetically distinct, dissemination-competent virus variants are rapidly selected from the viral quasispecies to be transmitted to vertebrates.

Palavras-Chave: Dengue virus type 2; American genotype Aedes aegypti; Midgut escape mutant; Amino acid substitution

Imprenta: Virology Journal, v. 10, n. 257, 2013

Identificador do objeto digital: 10.1186/1743-422X-10-257.

Descritores: Aedes aegypti - DNA ; Aedes aegypti - Flaviviridae ; Aedes aegypti - Genome ; Aedes aegypti - Pathogenesis ; Aedes aegypti - RNA ; Aedes aegypti - Virus ; Aedes aegypti - Dengue

Data de publicação: 2013