Expression of the herpes simplex virus type 2 glycoprotein D in baculovirus expression system and evaluation of its immunogenicity in guinea pigs.
Herpes simplex virus type 2 (HSV-2) is highly prevalent and major cause of genital herpes in humans. The life-long nature of infection and the increasing prevalence of genital herpes imply that vaccination is the best strategy for controlling the spread of infection and limiting HSV disease. HSV glycoprotein D (gD) is one of the most important viral immunogen which has an essential role in virus infectivity and induction of immune responses.



HSV-2 DNA was extracted and used as template in polymerase chain reactions to amplify gD2 gene. The PCR product was confirmed by restriction enzyme analysis, cloned into a cloning vector and then sequenced. The Bac-to-Bac expression system was used to express HSV-2 gD in insect cells. The expressed protein was used as subunit vaccine to immunize guinea pigs after confirmation.



The expressed protein was confirmed with SDS-PAGE and Western-blot analysis. In Western-blot analysis, two major protein bands, with approximate molecular weights of 52-55 and 41-43 kDa corresponding to the glycosylated and non-glycosylated forms of gD2 protein, were observed, respectively. Immunization with the recombinant gD2 could elicit humoral responses in guinea pigs as measured by neutralization test and ELISA, and offered high protection against induced HSV-2 genital disease.



The baculovirus expression of heterologous genes permits proper folding, post-translational modification and oligomerization in manners that are often identical to those that occur in mammalian cells. Expression of proteins under the control of the strong polyhedrin promoter, allowing high level protein production, can be used as subunit vaccine.
Fotouhi
Fatemeh
F
Department of Virology, School of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
Soleimanjahi
Hoorieh
H
Department of Virology, School of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
Roostaee
Mohammad Hasan
MH
Department of Virology, School of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
Dalimi Asl
Abdolhossein
A
Dept. of Parasitology, School of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
D000282:Administration, Intravaginal; D000818:Animals; D000914:Antibodies, Viral / Q000096:biosynthesis; D016367:Baculoviridae / Q000235:genetics* / Q000276:immunology; D002478:Cells, Cultured; D003001:Cloning, Molecular; D005260:Female; D006168:Guinea Pigs; D006367:HeLa Cells; D006558:Herpes Genitalis / Q000235:genetics / Q000276:immunology* / Q000517:prevention & control*; D022301:Herpes Simplex Virus Vaccines / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology; D018258:Herpesvirus 2, Human / Q000235:genetics* / Q000276:immunology*; D006801:Humans; D008297:Male; D018411:Spodoptera / Q000235:genetics / Q000276:immunology / Q000821:virology; D022223:Vaccines, Subunit / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology; D014759:Viral Envelope Proteins / Q000008:administration & dosage* / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology
D000914:Antibodies, Viral; D022301:Herpes Simplex Virus Vaccines; D022223:Vaccines, Subunit; D014759:Viral Envelope Proteins; C040570:glycoprotein D-herpes simplex virus type 2