Construction and characterization of human adenovirus serotype 3 packaged by serotype 7 hexon

Human adenovirus serotype 3 (Ad3) and serotype 7 (Ad7) are important pathogens causing respiratory tract diseases such as acute respiratory disease in pediatric and adult patients, but the immunodominant targets of Ad3- and Ad7-specific neutralizing antibodies (NAbs) remain unclear. A chimeric Ad vector, Ad3/H7, was constructed by replacing the Ad3 hexon gene (H3) with the hexon gene (H7) of Ad7. The chimeric viruses were successfully rescued in HEp-2 cells, and the Ad7 hexon was able to encapsidate the Ad3 genome, and functioned as efficiently as the Ad3 hexon. Furthermore, we tested the host neutralization responses against the viruses using BALB/C mice. Up to 97% of the NAbs produced by mice that were infected with these viruses were specific for the hexon protein in vitro. Preimmunization of mice with one of Ad7 and Ad3/H7 significantly prevented subsequent intranasal infection of the other type in vivo. In contrast, preimmunization of mice with one of Ad3 and Ad3/H7 did not remarkably prevent subsequent infection of the other type. We next evaluated the functional significance of hexon and other structural proteins specific NAbs to suppress the immunogenicity of Ad3/H3 and Ad3/H7 vectors expressing EGFP in mice preimmunized with wild type Ad. Preimmunization of mice with Ad7 evidently suppressed EGFP-specific humoral immune responses elicited by Ad3/H7, and did not exert suppressive effects on Ad3/H3. But contrary to the in vitro neutralization results, EGFP-specific humoral immune responses elicited by Ad3/H7 was remarkably inhibited in Ad3-preimmunization mice. The whole genome of the Ad7 strain was sequenced and aligned with Ad3. The major differences between Ad3 and Ad7 were only observed in the fiber and hexon among all structural proteins, and the variation between the hexons only located in four hypervariable regions (HVRs), HVR-1, -2, -5, and -7. These results thus suggest that Ad3- and Ad7-specific NAbs are directed primarily against the hexon proteins both in vitro and in vivo. But high titer Ad3 fiber-specific NAbs may also play an important role in blunting Ad3 immunogenicity in vivo. These studies contribute to a more profound understanding of Ad immunogenicity and have relevance for the design of novel Ad vaccine.

• We constructed a chimeric adenovirus vector A3/H7. • Ad3- and Ad7-specific NAbs are directed primarily against HVRs both in vitro and in vivo. • Ad3 fiber-specific NAbs also could suppress Ad3 immunogenicity in vivo.