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Vaccine Detail

BacHA derived from Influenza A virus (A/Indonesia/CDC669/2006(H5N1))
Vaccine Information
  • Vaccine Name: BacHA derived from Influenza A virus (A/Indonesia/CDC669/2006(H5N1))
  • Target Pathogen: Influenza virus
  • Target Disease: Influenza (flu)
  • Vaccine Ontology ID: VO_0004174
  • Type: Subunit vaccine
  • Status: Research
  • Antigen: Influenza A virus (A/Indonesia/CDC669/2006(H5N1)) HA hemagglutinin
  • HA gene engineering:
    • Type: Recombinant vector construction
    • Description: For the construction of recombinant baculovirus BacHA, the full-length open reading frame (ORF) of the HA gene (CDC/669/Indonesia/06 and CDC/594/Indonesia/06) was amplified and inserted into pFASTBacHT A (Invitrogen, San Diego, CA) using RsrII and HindIII restriction sites. The ie1 promoter was amplified from WSSV DNA using the primers WSSVie1F (5'-CCTACGTATCAATTTTATGTGGCTAATGGAGA-3') and WSSVie1R (5'-CGCGTCGACCTTGAGTGGAGAGAGAGCTAGTTATAA-3') and then inserted into pFASTBacHT A using AccI and RsrII restriction sites. For the generation of recombinant baculoviruses, the constructs were integrated into the baculovirus genome within DH10Bac (Invitrogen) through site-specific transposition using Bac-To-Bac system (Invitrogen). The recombinant bacmids were then transfected into Sf9 cells, and the budded virus particles released into the medium were harvested at 4 days posttransfection (Prabakaran et al., 2010).
    • Detailed Gene Information: Click Here.
  • HA gene engineering:
    • Type: Recombinant vector construction
    • Description: For the construction of recombinant baculovirus BacHA, the full-length open reading frame (ORF) of the HA gene (CDC/669/Indonesia/06 and CDC/594/Indonesia/06) was amplified and inserted into pFASTBacHT A (Invitrogen, San Diego, CA) using RsrII and HindIII restriction sites. The ie1 promoter was amplified from WSSV DNA using the primers WSSVie1F (5'-CCTACGTATCAATTTTATGTGGCTAATGGAGA-3') and WSSVie1R (5'-CGCGTCGACCTTGAGTGGAGAGAGAGCTAGTTATAA-3') and then inserted into pFASTBacHT A using AccI and RsrII restriction sites. For the generation of recombinant baculoviruses, the constructs were integrated into the baculovirus genome within DH10Bac (Invitrogen) through site-specific transposition using Bac-To-Bac system (Invitrogen). The recombinant bacmids were then transfected into Sf9 cells, and the budded virus particles released into the medium were harvested at 4 days posttransfection (Prabakaran et al., 2010).
    • Detailed Gene Information: Click Here.
  • Adjuvant: cholera toxin B subunit
  • Immunization Route: Oral
Host Response

Mouse Response

  • Host Strain: BALB/c
  • Vaccination Protocol: Prior to immunization, all mice were starved for 2 h; otherwise food and water were supplied ad libitum. Thirty mice per each experimental group (n = 30/group) were immunized intragastrically by oral gavage on days 0, 7, and 21 with 200 µl containing inactivated or live recombinant baculovirus vaccine at a log2 HA titer of 8 suspended in phosphate-buffered saline (PBS), pH 7.4, either adjuvanted with 10 µg rCTB or unadjvanted (Prabakaran et al., 2010).
  • Challenge Protocol: Six mice per group were challenged intranasally with 5 50% mouse lethal doses (MLD50) of homologous (CDC/669/Indonesia/06 clade 2.1) and heterologous (Vietnam/1203/2004 clade 1.0) HPAI H5N1 virus strains. The MLD50 of the influenza virus required for intranasal challenge experiments was predetermined. To determine the effect of adjuvant efficacy, animals immunized with vaccines without adjuvant or only with rCTB were also maintained as control groups. Mice were observed daily to monitor body weight and mortality (Prabakaran et al., 2010).
  • Efficacy: Viral challenge studies showed that live BacHA derived from Influenza A virus (A/Indonesia/CDC669/2006(H5N1)) was able to provide 100% protection against 5 50% mouse lethal doses (MLD(50)) of homologous (clade 2.1) and heterologous (clade 1) H5N1 (Prabakaran et al., 2010).
References
Prabakaran et al., 2010: Prabakaran M, Madhan S, Prabhu N, Qiang J, Kwang J. Gastrointestinal delivery of baculovirus displaying influenza virus hemagglutinin protects mice against heterologous H5N1 infection. Journal of virology. 2010; 84(7); 3201-3209. [PubMed: 20071572].