A Recombinant Influenza A/H1N1 Carrying A Short Immunogenic Peptide of MERS-CoV as Bivalent Vaccine in BALB/c Mice

Shehata, Mahmoud M.; Kandeil, Ahmed; Mostafa, Ahmed; Mahmoud, Sara H.; Gomaa, Mokhtar R.; El-Shesheny, Rabeh; Webby, Richard; Kayali, Ghazi; A. Ali, Mohamed

The short immunogenic peptide 736 CALPDTPSTLTPRSVRSVPGEMRLA 761 from MERS-CoV spike glycoprotein was inserted into the stalk region of NA gene of influenza A/California/04/2009 (H1N1) (H1N1pdm09) virus using the designed primers. A new recombinant H1N1 NA carrying short peptide of MERS-CoV (chimeric bivalent 5 + 3) was successfully constructed by recombinant ligation and cloning in pHW2000 vector ( Figure 1A ). Using the eight-plasmid reverse genetics system, recombinant 5 + 3 rgH1N1 (Control virus), and chimeric bivalent 5 + 3 (candidate bivalent virus) were generated. The reverse genetic system to generate the chimeric bivalent virus is illustrated in Figure 1B . The rescued bivalent and control viruses were propagated in SPF embryonated chicken eggs for three passages and were safe to embryos (no mortality) at 48 h post infection (pi). The inserted sequence for the immunogenic peptide of MERS-CoV spike glycoprotein was stable without any associated adaptive mutations/deletions in the NA gene during three successive passages in eggs.