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. 2004 Jun 26;363(9427):2122-7.
doi: 10.1016/S0140-6736(04)16501-X.

Mucosal immunisation of African green monkeys (Cercopithecus aethiops) with an attenuated parainfluenza virus expressing the SARS coronavirus spike protein for the prevention of SARS

Alexander Bukreyev  1 , Elaine W Lamirande, Ursula J Buchholz, Leatrice N Vogel, William R Elkins, Marisa St Claire, Brian R Murphy, Kanta Subbarao, Peter L Collins
Affiliations

Mucosal immunisation of African green monkeys (Cercopithecus aethiops) with an attenuated parainfluenza virus expressing the SARS coronavirus spike protein for the prevention of SARS

Alexander Bukreyev et al. Lancet. .

Abstract

Background: The outbreak of severe acute respiratory syndrome (SARS) in 2002 was caused by a previously unknown coronavirus-SARS coronavirus (SARS-CoV). We have developed an experimental SARS vaccine for direct immunisation of the respiratory tract, the major site of SARS- coronavirus transmission and disease.

Methods: We expressed the complete SARS coronavirus envelope spike (S) protein from a recombinant attenuated parainfluenza virus (BHPIV3) that is being developed as a live attenuated, intranasal paediatric vaccine against human parainfluenza virus type 3 (HPIV3). We immunised eight African green monkeys, four with a single dose of BHPIV3/ SARS-S and four with a control, BHPIV3/Ctrl, administered via the respiratory tract. A SARS-coronavirus challenge was given to all monkeys 28 days after immunisation.

Findings: Immunisation of animals with BHPIV3/SARS-S induced the production of SARS-coronavirus-neutralising serum antibodies, indicating that a systemic immune response resulted from mucosal immunisation. After challenge with SARS coronavirus, all monkeys in the control group shed SARS coronavirus, with shedding lasting 5-8 days. No viral shedding occurred in the group immunised with BHPIV3/SARS-S.

Interpretation: A vectored mucosal vaccine expressing the SARS-coronavirus S protein alone may be highly effective in a single-dose format for the prevention of SARS.

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Figures

Figure 1
Figure 1
RNA genome maps for BHPIV3/SARS-S and control BHPIV3/Ctrl viruses N=nucleocapsid protein. P=phosphoprotein. F=fusion glycoprotein. HN=haemagglutinin-neuraminidase glycoprotein. L=polymerase protein. Black bars at beginning and end of each gene represent PIV3-specific transcription signals, and gap between rectangles represents the PIV3 intergenic trinucleotide. Leader (Le) and trailer (Tr) sequences at 3′ and 5′ ends of the PIV3 genome are short extragenic regions containing promoter sequences.
Figure 2
Figure 2
Expression of SARS-coronavirus S protein by BHPIV3/SARS-S in cell culture and its absence in the BHPIV3 virus particle A: western blot analysis shows presence of the SARS-coronavirus S protein in lysates of cells infected with BHPIV3/SARS-S (lane 3) and its absence in lysates from cells infected with BHPIV3/Ctrl (lane 4). S protein was not detectable in purified, concentrated BHPIV3/SARS-S virus particles (lane 1) or in the negative control BHPIV3/Ctrl (lane 2). B: direct Coomassie staining of a gel loaded with replicate samples of purified, concentrated BHPIV3/Ctrl (lane 1) and BHPIV3/SARS-S (lane 2) shows viral protein was present in excess of what should be necessary for detection by western blot analysis.
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