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

Mapping the epitope of neutralizing monoclonal antibodies against human adenovirus type 3

“…The following strains of adenoviruses used in this study were obtained as previously described (Tian et al, 2011;Qiu et al, 2012;Zhang et al, 2009): recombinant adenovirus rAd3EGFP encoding a HAdV-3 GZ-01 genome and an enhanced green fluorescent protein (eGFP) with an E3 region deletion, hexon chimeric adenovirus rAd3egf/H7 generated by replacing the HAdV-3 hexon gene of the rAd3EGFP with the hexon gene from HAdV-7 GZ08 strain, epitope mutants (rAd3H7R1, rAd3H7R2, rAd3H7R5, and rAd3H7R7) from rAd3egf/H7 replaced with corresponding HAdV-3 epitopes, and epitope chimeric mutant rAdMHE3 from rAd3EGFP replaced with HVR5 of HAdV-7. All the adenoviruses were cultured in HEp-2 cells or AD293 cells as previously described (Tian et al, 2011).…”

“…All the adenoviruses were cultured in HEp-2 cells or AD293 cells as previously described (Tian et al, 2011). Adenovirus particles were purified by standard CsCl gradient centrifugation as previously described (Wu et al, 2002).…”

“…Adenovirus particles were purified by standard CsCl gradient centrifugation as previously described (Wu et al, 2002). The virus particle (VP) titers were determined by spectrophotometry using a conversion factor of 1.1 × 10 12 VPs per absorbance unit at 260 nm (Tian et al, 2011).…”

“…DQ099432) and eGFP with an E3 region deletion was constructed, as previously described . The hexon-chimeric adenovirus vector pAd3egf/H7, in which the HAdV-3 hexon gene in the pBRAd E3GFP vector was replaced with the hexon gene from HAdV-7, was constructed as previously described (Tian et al, 2011). The shuttle vector pBRLR was also constructed as previously described (Qiu et al, 2012).…”

“…The adenovirus capsid icosahedron is composed of three major structural proteins: hexon, penton base, and fiber. The hexon protein is the major antigenic determinant recognized by neutralizing antibodies (NAbs) (Tian et al, 2011;Yu et al, 2013;Wu et al, 2002). Type-specific epitopes on hexons have been proposed to reside within seven highly variable regions (HVRs), of which HVR7 can be further subdivided into three http://dx.doi.org/10.1016/j.virusres.2015.06.002 0168-1702/© 2015 Elsevier B.V. All rights reserved.…”