• Vaccine Ontology ID: VO_0011561
• Type: Recombinant vector vaccine
• Status: Research
• Antigen: Andes virus nucleocapsid protein (ANDVsSgp1)
• ANDVsSgp1 gene engineering:
  • Type: Recombinant vector construction
  • Description: Nonreplicating (E1− E3−) Ad vectors expressing ANDV proteins were constructed using the AdMax HI-IQ system (Microbix, Toronto, Canada). The ANDV GN, GC, GPC, and N open reading frames were PCR amplified from plasmids containing the appropriate cDNAs derived from a Chilean ANDV isolate, strain 9717869. AdEmpty was constructed using plasmid pDC316(io) containing no ANDV sequences. These pDC316(io)-based ANDV plasmids were cotransfected with plasmid pBHGloxΔE1,3Cre, containing the remainder of the Ad5 genomic plasmid, into 293 IQ cells. Supernatants were collected after 6 to 8 days, and the presence of Ad vectors expressing the ANDV proteins (designated AdN, AdGN, AdGC, and AdGPC) was confirmed by immunoprecipitation. Ad vectors were plaque purified, propagated in large-scale infections in 293 IQ cells, and purified using standard CsCl gradient methods (Safronetz et al., 2009).
  • Detailed Gene Information: Click Here.
• Vector: Nonreplicating adenovirus (Ad) vectors
• Immunization Route: Intraperitoneal injection (i.p.)

Hamster Response

• Host Strain: Syrian gold
• Vaccination Protocol: Syrian golden hamsters (Mesocricetus auratus) (4- to 6-week-old males; Charles River, Pointe Claire, Canada) were group housed in microisolator units situated in the biosafety level 4 area of the National Microbiology Laboratory, Public Health Agency of Canada. Prior to vaccine experiments, the 50% lethal dose (LD50) for i.p. injections of ANDV in hamsters was established by inoculating anesthetized animals with 0.8 to 80,000 FFU (using 10-fold dilutions) of ANDV. Hamsters were monitored twice daily for clinical signs of illness according to an approved scoring sheet (ruffled fur, lethargy, inappetence, and labored breathing). For vaccination, hamsters were anesthetized with isoflurane, a preimmunization blood sample was collected, and the animals were immunized with the Ad vectors using 108 (293 cell) PFU of each vector diluted in 100 μl phosphate-buffered saline at two sites in the hind-leg musculature (Safronetz et al., 2009).
• Challenge Protocol: After 28 days, a second blood sample was collected and hamsters were challenged with ANDV by i.p. injection of 100 LD50s (equivalent to 154 FFU). Hamsters were examined twice daily for signs of illness. Survivors were monitored for 40 to 45 days and then anesthetized and exsanguinated via cardiac puncture (Safronetz et al., 2009).
• Efficacy: Nonreplicating adenovirus (Ad) vectors that express Andes hantavirus (ANDV) nucleocapsid protein (AdN, ANDVsSgp1) or glycoproteins (AdG(N) and AdG(C)) were constructed . When administered once, all three Ad vectors, individually or in combination, elicited a robust immune response that protected Syrian hamsters from a lethal ANDV infection that mimics the pulmonary disease seen in humans. No vaccinated animal died, and there were no obvious clinical signs of disease (Safronetz et al., 2009).