RESUMEN
Epstein-Barr virus (EBV) associated with several diseases such as contagious mononucleosis chronic active EBV infection, and diverse sorts of malignant tumors. Therefore, using applicable vaccines could be advantageous for public health. Yet, the vaccine has been unavailable to protect from EBV so far. In the current study, to develop a multi-peptide vaccine for EBV and assess its expression in Pichia pastoris yeast system, three immunodominant sequences in glycoprotein (gp) 85, gp350 and latent membrane protein 1 (LMP1) were chosen. To construct fusion peptide, -GGGGS- liker was applied. After cloning the fusion peptide in the pPICZαA expression vector, this recombinant vector processed and transfected into Pichia pastoris host cells. The expression of high level of EBV fusion peptide was confirmed by dot blot and SDS-PAGE procedures. The Pichia pastoris is capable of supporting EBV fusion peptide expression. The application of this fusion peptide as a peptide vaccine to fight EBV is suggested.
Asunto(s)
Herpesvirus Humano 4/inmunología , Fragmentos Fc de Inmunoglobulinas/genética , Glicoproteínas de Membrana/genética , Proteínas del Envoltorio Viral/genética , Proteínas de la Matriz Viral/genética , Vacunas Virales/biosíntesis , Secuencia de Aminoácidos , Linfoma de Burkitt/inmunología , Linfoma de Burkitt/prevención & control , Linfoma de Burkitt/virología , Clonación Molecular , Expresión Génica , Vectores Genéticos/química , Vectores Genéticos/metabolismo , Herpesvirus Humano 4/genética , Humanos , Fragmentos Fc de Inmunoglobulinas/inmunología , Mononucleosis Infecciosa/inmunología , Mononucleosis Infecciosa/prevención & control , Mononucleosis Infecciosa/virología , Glicoproteínas de Membrana/inmunología , Péptidos/genética , Péptidos/inmunología , Pichia/genética , Pichia/metabolismo , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/inmunología , Vacunas de Subunidad , Proteínas del Envoltorio Viral/inmunología , Proteínas de la Matriz Viral/inmunologíaRESUMEN
OBJECTIVES: Human Cytomegalovirus (HCMV) remains a major morbidity and mortality cause in immuno suppressed patients. Therefore, significant effort has been made towards the development of a vaccine. In this study, the expression of the pp65 and gB fusion peptides and Fc domain of mouse IgG2a as a novel delivery system for selective uptake of antigens by antigen-presenting cells (APCs) in Pichia pastoris yeast system were studied. MATERIALS AND METHOD: In this study, four immune dominant sequences in pp65 protein and 3 immuno dominant sequences in gB protein were selected according to literature review. Peptide linker -GGGGS- was used for construction of fusion peptide. This fusion peptide was cloned in the pPICZαA expression vector and transfected into P. pastoris host cells. RESULTS: Dot blot and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) techniques showed that a high level of pp65-gB-Fc fusion peptide was expressed. CONCLUSION: This CMV pp65-gB-Fc fusion peptide could be a promising candidate for the development of a novel peptide vaccine.
