RESUMEN
AIM: To develop a recombinant protein vaccine with epitopes on L1 protein for prevention of HPV16 infection. METHODS: One B cell epitope and one T cell epitope from HPV16 L1 protein were chosen to construct genes encoding different recombinant proteins. Then all the 3D structures of the proteins were predicted by Geno3D in internet. An optimal protein named RH was chosen in which three B epitopes' location simulates that in the real HPV16 L1. Gene X (consisting of B+T+T) and gene Y (consisting of B+T) were constructed by multiple PCR. RH gene (consisting of X+X+X+X+Y+Y+Y) was constructed, sub-cloned into pFastbac1, and expressed in Sf9. RESULTS: RH protein with epitopes on HPV16 L1 was designed and predicted by Geno3D. RH gene was constructed and sub cloned into pFastbac1. Finally, it was successfully expressed in baculovirus expression system. CONCLUSION: A HPV16 recombinant vaccine is successfully constructed and expressed.
Asunto(s)
Proteínas de la Cápside/genética , Diseño Asistido por Computadora , Proteínas Oncogénicas Virales/genética , Vacunas contra Papillomavirus/genética , Ingeniería de Proteínas , Animales , Proteínas de la Cápside/química , Proteínas de la Cápside/metabolismo , Línea Celular , Expresión Génica , Proteínas Oncogénicas Virales/química , Proteínas Oncogénicas Virales/metabolismo , Vacunas contra Papillomavirus/química , Vacunas contra Papillomavirus/metabolismo , Estructura Terciaria de ProteínaRESUMEN
AIM: To establish a set of reasonable and convenient experimental system to provide a screening method for the development of novel immunoregulatory oligodeoxynucleotides. METHODS: The human PBMCs were stimulated by CpG ODN and/or immunoregulatory ODN. The cell proliferation and anti-viral activity of the supernatant induced by CpG ODN were examined by thymidine incorporation and anti-viral bioassay to evaluate the immunoregulatory activity of candidate ODN. The experimental conditions were also optimized. RESULTS: A screening method on which A151, a positive immunoregulatory ODN, inhibited the proliferation and anti-viral activity of the CpG ODN-induced human PBMCs was successfully established. CONCLUSION: The successful establishment of CpG ODN based screening method lays the foundations for further development of novel immunoregulatory oligodeoxynucleotides.