title: [Construction of a recombinant eukaryotic expression vector containing HSV 2gD  and its  immune  responses].
abstract: OBJECTIVE
  To construct and express pcDNA3.1-IL-18-HSV P6 recombinant DNA vaccine, and to observe the immune responses of pcDNA3.1-IL-18-HSV P6 genetic vaccine.
  
  
  METHODS
  Genes of P6 and IL-18 were cloned into the eukaryotic expression vector pcDNA3.1(-) respectively, and confirmed by enzyme digestion and sequence analysis, the recombinant plasmid pcDNA3.1-IL-18-HSVP6 was transformed into CHO cells. The expressed protein was characterized by indirect immunofluorescence and Western blotting. The recombinant plasmid pcDNA3.1-IL-18-HSVP6 was used to inoculate BALB/c mice by intramuscular injection for three times, once a week. One week after the last vaccination, the levels of specific IgG antibody, IFN-γ and IL-18 were detected by ELISA; One month after the last vaccination, spleen cells of vaccinated mice were separated and proliferation of spleen lymphocytes were detected by MTT assay.
  
  
  RESULTS
  Recombinant pcDNA3.1-IL-18-HSV P6 plasmid was confirmed by enzyme digestion and DNA sequencing. After inoculated by pcDNA3.1-IL-18-HSV P6 vaccine, the mice could produce higher level of splenocytes proliferation and secrected higher level of IFN-γ, IL-18 and specific antibody.
  
  
  CONCLUSIONS
  DNA vaccine of pcDNA3.1-IL-18-HSV P6 has been successfully constructed, and it can effectively induce humoral and cellular immune responses, which provided a basis for constructing new type of DNA vaccine.
authors.affiliation: Department of Aetiology, Taishan Medical University, Taian, China. fpjiao@tsmc.edu.cn
authors.forename: Feng-ping
authors.initials: FP
authors.lastname: Jiao
authors.forename: Li
authors.initials: L
authors.lastname: Liu
authors.forename: Yu
authors.initials: Y
authors.lastname: Wang
authors.forename: Ai-lian
authors.initials: AL
authors.lastname: Yu
authors.forename: Guang-fu
authors.initials: GF
authors.lastname: Yu
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D005260:Female; D005822:Genetic Vectors; D007074:Immunoglobulin G / Q000097:blood; D007371:Interferon-gamma / Q000097:blood; D020382:Interleukin-18 / Q000097:blood; D051379:Mice; D008807:Mice, Inbred BALB C; D016133:Polymerase Chain Reaction; D014759:Viral Envelope Proteins / Q000235:genetics*
chemical_list: D000914:Antibodies, Viral; D007074:Immunoglobulin G; D020382:Interleukin-18; D014759:Viral Envelope Proteins; C040570:glycoprotein D-herpes simplex virus type 2; D007371:Interferon-gamma