• Vaccine Name: Seoul virus vaccine rCAV-2-Gc
• Target Pathogen: Hantavirus
• Target Disease: Hantavirus Pulmonary Syndrome
• Vaccine Ontology ID: VO_0011402
• Type: Recombinant vector vaccine
• Status: Research
• Antigen: Seoul virus Gc glycoprotein
• Gc gene engineering:
  • Type: Recombinant protein preparation
  • Description: The Gc protein expressed by rCAV-2-Gc in MDCK cells was evaluated by a SEOV-specific indirect IFA. MDCK cells grown on 15 mm glass coverslips in 12-well culture plates were infected with rCAV-2-Gc or CAV-2 at an m.o.i. of 20. After 48 h infection, the coverslips were rinsed once with PBS (pH 7.4), fixed with acetone for 10 min at room temperature and then reacted with rabbit anti-SEOV polyclonal antiserum and washed three times with PBS. The fixed monolayers were incubated at 37 °C for 30 min in a moist chamber with donkey anti-rabbit IgG labelled with fluorescence isothiocyanate (Amersham). The coverslips were rinsed three times with PBS. Cell monolayers that bound the antibody were covered with glycerine and examined for specific fluorescence under a Zeiss Axioplan fluorescence microscope. Expression of SEOV Gc in MDCK cells infected with rCAV-2-Gc was identified by Western blotting. rCAV-2-Gc-infected cell lysates were separated by 12 % SDS-PAGE, and the proteins transferred to nitrocellulose membrane (Pall Corporation) and probed with positive serum against SEOV and HRP-labelled goat anti-mouse IgG antibody (Sigma) (Yuan et al., 2010).
  • Detailed Gene Information: Click Here.
• Immunization Route: Intramuscular injection (i.m.)

Mouse Response

• Host Strain: BALB/c
• Vaccination Protocol: Mice were randomly assigned to four experimental groups (20 mice per group). Group I was intramuscularly inoculated once with 0.1 ml rCAV-2-Gc (108.0 p.f.u. ml–1); group II received 0.1 ml CAV-2 (108.2 p.f.u. ml–1) intramuscularly as a negative control; group III were inoculated intramuscularly with one dose of HFRS bivalent purified vaccine Youerjian (0.5 ml per dose; GuangDong HongMing Biological Science and Technology Co.) as a positive-control; and group IV were injected with 0.1 ml PBS as a negative control (Yuan et al., 2010).
• Challenge Protocol: At the end of the trial, all mice were injected intramuscularly with SEOV strain CC-2 diluted in 0.2 ml PBS. The challenge dose for each virus was 2000 p.f.u. This dose is 1000 50 % infective doses for SEOV. At 14 days after challenge, the mice were sacrificed by CO2 asphyxiation, as approved by the China Small Animal Protection Association. Pre- and post-challenge sera were evaluated for the presence of N-specific antibodies by ELISA and for the presence of neutralizing antibodies by FRNT. Detecting post-challenge N-specific antibody indicated that the mice were infected with the challenge virus (Yuan et al., 2010).
• Efficacy: eplication-competent recombinant canine adenovirus type 2 expressing the Gc protein of SEOV (rCAV-2-Gc) in BALB/c mice induced complete protection against a intensive infectious challenge with ~1,000 50% infective doses (ID50) for SEOV strain CC-2 (Yuan et al., 2010).