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. 2013 Jun 18;8(6):e66276. doi: 10.1371/journal.pone.0066276

Figure 1. Recombinant adenoviruses carrying codon-optimized HA genes expressed HA proteins and elicited neutralizing antibodies against H5N1 viruses in rhesus macaques.

Figure 1

(A) Schematic representation of adenoviral vectors carrying H5N1 influenza HA gene (Ad5-HA). The expression cassette, containing the HA gene under the control of the CMV promoter and BGH polyA signal, was cloned into the adenoviral E1 region. (B) Expression of the HA protein mediated by recombinant adenovirus Ad5-HAs. The HA proteins before cleavage (H0) and after cleavage (HA1 and HA2) were detected using chicken serum against HA in Western blots. The Vero cells were harvested for analysis at 48 hours after infection with Ad-Empty, Ad5-HA04VN, Ad5-HA97HK, and Ad5-HA06ZJ at MOIs of 10. (C) Dynamics of the antibody response in a Chinese rhesus macaque immunized with Ad5-HAs. The sera were collected at 2-week intervals. The neutralizing titers against the recombinant H5N1 viruses (r97HKHA-PR8, r04VNHA-PR8, and r06ZJHA-PR8) were determined using a micro-neutralization assay. The arrows indicate the time of immunization. (D) The binding specificity of the sera against HA of different clades expressed by the recombinant influenza viruses was determined by Western blot analysis.