Development of Replication-Defective Adenovirus Serotype 5 Containing the Capsid and 3C Protease Coding Regions of Foot-and-Mouth Disease Virus as a Vaccine Candidate

Abstract

A recombinant replication-defective human adenovirus serotype 5 vector containing FMDV capsid, P1-2A, and viral 3C protease coding regions was constructed. Two viral clones were isolated, Ad5-P12X3CWT, containing the wild-type (WT) 3C protease that processes capsid polyprotein precursor into mature capsid proteins, and Ad5-P12X3CMUT, containing a point mutation in the protease coding region that inhibits processing. In 293 cells infected with either virus, synthesis of the FMDV capsid polyprotein precursor occurred, but processing of the polyprotein into structural proteins VP0, VP3, and VP1 occurred only in 3CWT virus-infected cells. Immunoprecipitation with monospecific and monoclonal antibodies indicates possible higher order structure formation in Ad5-P12X3CWT virus-infected cells. The viruses were used to elicit immune responses in mice inoculated intramuscularly (im). Only virus containing the 3CWT elicited a neutralizing antibody response. After boosting, this neutralizing antibody response increased. Swine inoculated im with Ad5-P12X3CWT virus developed a neutralizing antibody response and were either completely or partially protected from contact challenge with an animal directly inoculated with virulent FMDV. This adenovirus vector may be an efficient system for the delivery of FMDV cDNA into animals, leading to a high level of neutralizing antibody production and protection from FMDV challenge.

Constitutive Expression of Alpha Interferon by Skin Dendritic Cells Confers Resistance to Infection by Foot-and-Mouth Disease Virus

“…Levels of viremia peak between 2 and 3 days after infection, and there is a rapid development of antibodies correlating with viral clearance. Consistent with this, the presence of neutralizing antibodies correlates with protection against reinfection and is believed to be the important effector function in vaccinated animals (31,33,52). However, the role of innate and cellular immune responses in the protection induced by vaccines is not understood.…”

Granulocyte-macrophage colony-stimulating factor does not increase the potency or efficacy of a foot-and-mouth disease virus subunit vaccine

“…Recombinant virus Ad5-pGMCSF was produced by transfection of 293 cells with PacI digested pAd5-pGMCSF following the protocol described (Wu et al, 2003b). The virus was isolated, propagated in 293 cells, and purified by CsCl gradient centrifugation (Mayr et al, 1999;Moraes et al, 2002).…”

“…Plasmid pP12X3C (Mayr et al, 1999), which contains the complete P1-2A and 3C coding regions and partial 2B and 3B coding regions of FMDV A12, was digested with the same enzymes to remove the A12 P1-2A coding region and ligated to O1C P1-2A to generate pP1-2A(O1C)X3C. This plasmid was digested with BglII and XbaI to remove the O1C P1-2A and A12 3C coding regions and ligated to similarly digested Ad5 transfer vector pShuttle (He et al, 1998).…”

“…Human adenovirus (Ad5) has been used as a vector for FMD vaccines encoding the capsid (P1-2A) (Sanz-Parra et al 1999a,b) or P1-2A and 3C protease coding regions of FMDV (Mayr et al 1999, Moraes et al 2002, Wu et al 2003b. Inoculation with one dose of Ad5 containing the serotypes A24 Cruzeiro P1-2A and A12 3C coding regions (Ad5-A24) protected swine from direct inoculation challenge with homologous virus (Moraes et al 2002).…”

Induction of mucosal immune responses and protection of cattle against direct-contact challenge by intranasal delivery with foot-and-mouth disease virus antigen mediated by nanoparticles

“…However, the construct lacked the coding regions for most other nonstructural proteins. 54,55 TEM showed that cells transfected with plasmid pcDNA3.1/P12A3C had detectable empty capsid structures that contained all or most epitopes and induced both humoral and cell-mediated immune responses. We detected isotype-specific sIgA in nasal mucosa samples, and IgA and IgG responses in serum after vaccination of cattle at different time points.…”