title: Characterization of the immune responses elicited by baculovirus-based vector vaccines against influenza virus hemagglutinin.
abstract: OBJECTIVE
  To compare the specific immune responses elicited by different baculovirus vectors in immunized mice.
  
  
  METHODS
  We constructed and characterized two recombinant baculoviruses carrying the expression cassette for the H5N1 influenza virus hemagglutinin (HA) gene driven by either an insect cell promoter (vAc-HA) or a dual-promoter active both in insect and mammalian cells (vAc-HA-DUAL). Virus without the HA gene (vAc-EGFP) was used as a control. These viruses were used to immunize mice subcutaneously and intraperitoneally. The production of total and specific antibodies was determined by ELISA and competitive ELISA. Cytokine production by the spleen cells of immunized mice was studied using the ELISPOT assay.
  
  
  RESULTS
  Both the vAc-HA and vAc-HA-DUAL vectors expressed HA proteins in insect Sf9 cells, and HA antigen was present in progeny virions. The vAc-HA-DUAL vector also mediated HA expression in virus-transduced mammalian cell lines (BHK and A547). Both vAc-HA and vAc-HA-DUAL exhibited higher transduction efficiencies than vAc-EGFP in mammalian cells, as shown by the expression of the reporter gene egfp. Additionally, both vAc-HA and vAc-HA-DUAL induced high levels of HA-specific antibody production in immunized mice; vAc-HA-DUAL was more efficient in inducing IFN-γ and IL-2 upon stimulation with specific antigen, whereas vAc-HA was more efficient in inducing IL-4 and IL-6.
  
  
  CONCLUSIONS
  Baculovirus vectors elicited efficient, specific immune responses in immunized mice. The vector displaying the HA antigen on the virion surface (vAc-HA) elicited a Th2-biased immune response, whereas the vector displaying HA and mediating HA expression in the cell (vAc-HA-DUAL) elicited a Th1-biased immune response.
authors.affiliation: Department of Microbiology and Microbial Engineering, School of Life Sciences, Fudan University, Shanghai, China.
authors.forename: Zhi-peng
authors.initials: ZP
authors.lastname: Hu
authors.forename: Juan
authors.initials: J
authors.lastname: Yin
authors.forename: Yuan-yuan
authors.initials: YY
authors.lastname: Zhang
authors.forename: Shu-ya
authors.initials: SY
authors.lastname: Jia
authors.forename: Zuo-jia
authors.initials: ZJ
authors.lastname: Chen
authors.forename: Jiang
authors.initials: J
authors.lastname: Zhong
mesh_terms: D000818:Animals; D000917:Antibody Formation; D016367:Baculoviridae / Q000235:genetics*; D002460:Cell Line; D005260:Female; D005822:Genetic Vectors / Q000235:genetics; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000235:genetics / Q000276:immunology / Q000627:therapeutic use*; D053124:Influenza A Virus, H5N1 Subtype / Q000235:genetics / Q000276:immunology*; D007252:Influenza Vaccines / Q000235:genetics* / Q000276:immunology / Q000627:therapeutic use*; D007371:Interferon-gamma / Q000276:immunology; D007378:Interleukins / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D009976:Orthomyxoviridae Infections / Q000276:immunology / Q000517:prevention & control*; D011995:Recombination, Genetic; D014161:Transduction, Genetic; D014611:Vaccination
chemical_list: D019267:Hemagglutinin Glycoproteins, Influenza Virus; D007252:Influenza Vaccines; D007378:Interleukins; D007371:Interferon-gamma