RESUMEN

Two human papillomavirus (HPV) prophylactic vaccines are currently available in the market: Gardasil and Cervarix. These two vaccines work against tumor high-risk subtypes HPV 16 and HPV 18. However, they do not include other high-risk subtypes such as HPV 58. Epidemiological research in China shows that HPV 58 is a prevalent high-risk subtype, second only to HPV 16 and HPV 18. Thus, for cervical cancer prevention in China, developing a vaccine against HPV 58 is necessary. In this study, HPV 58 virus-like particles (VLPs) were expressed in the Pichia pastoris, and subsequently purified through pretreatment and a three-step purification process consisting of strong cation exchange chromatography, size-exclusion chromatography, and hydroxyapatite chromatography. The highly purified HPV 58 VLPs were confirmed by sodium dodecyl sulfate polyacrylamide gel electrophoresis, electron microscopy, dynamic laser scattering, and ultracentrifugation. The purified VLPs were used to immunize mice to test their ability to induce humoral immunity. Enzyme-linked immunosorbent assays were performed on the sera of the immunized mice and significantly high anti-HPV 58 VLP antibody titers were observed. The immunogenicity study demonstrates that the purified HPV 58 VLPs are HPV vaccine candidates.

Asunto(s)

Alphapapillomavirus/inmunología , Proteínas de la Cápside/inmunología , Vacunas contra Papillomavirus/inmunología , Pichia/metabolismo , Vacunas de Partículas Similares a Virus/inmunología , Alphapapillomavirus/genética , Animales , Western Blotting , Proteínas de la Cápside/genética , Proteínas de la Cápside/aislamiento & purificación , Proteínas de la Cápside/metabolismo , Cromatografía en Gel , Electroforesis en Gel de Poliacrilamida , Femenino , Fermentación , Genes Virales , Inmunidad Humoral , Ratones , Ratones Endogámicos BALB C , Microscopía Electrónica , Pichia/genética , Plásmidos/genética , Plásmidos/metabolismo , Transformación Genética , Ultracentrifugación , Vacunación