Autor(es): Noranate Nitchakarn, Takeda Naokazu, Chetanachan Prukswan, Sittisaman Pathompong, A-Nuegoonpipat Atchareeya, Anantapreecha Surapee

Resumo: Chikungunya virus (CHIKV) is becoming a global concern due to the increasing number of outbreaks throughout the world - the absence of any CHIKV-specific vaccine or treatment. Virus-like particles (VLPs) are multistructured proteins that mimic the organization - conformation of native viruses but lack the viral genome. They are noninfectious - potentially safer vaccine c-idates. Recent studies demonstrated that the yield of CHIKV VLPs varies depending on the strains, despite the 95% amino acid similarity of the strains. This might be due to the codon usage, since protein expression is differently controlled by different organisms. We optimized the region encoding CHIKV structural proteins, C-E3-E2-6k-E1, inserted it into a mammalian expression vector, - used the resulting construct to transfect 293 cells. We detected 50-kDa proteins corresponding to E1 -/or E2 in the cell lysate - the supernatant. Transmission electron microscopy revealed spherical particles with a 50- to 60-nm diameter in the supernatant that resembled the native CHIKV virions. The buoyant density of the VLPs was 1.23 g/mL, - the yield was 20 µg purified VLPs per 108 cells. The VLPs aggregated when mixed with convalescent sera from chikungunya patients, indicating that their antigenicity is similar to that of native CHIKV. Antibodies elicited with the VLPs were capable of detecting native CHIKV, demonstrating that the VLPs retain immunogenicity similar to that of the native virion. These results indicated that CHIKV VLPs are morphologically, antigenically, - immunologically similar to the native CHIKV, suggesting that they have potential for use in chikungunya vaccines.

Imprenta: PloS One, v. 9, n. 9, p. e108169, 2014

Identificador do Objeto Digital: 10.1371/journal.pone.0108169

Descritores: Chikungunya virus - Cell ; Chikungunya virus - DNA ; Chikungunya virus - Genome ; Chikungunya virus - Molecular structure ; Chikungunya virus - Pathogenesis ; Chikungunya virus - Proteins ; Chikungunya virus - Antibodies ; Chikungunya virus - Viral infections ; Chikungunya Virus - Virus ; Chikungunya virus - Transmission ; Chikungunya virus - Vaccine ; Chikungunya virus - Immunology ; Chikungunya virus - Public health

Data de Publicação: 2014