title: Construction and characterization of a recombinant pseudorabies virus expressing porcine reproductive and respiratory syndrome virus GP5.
abstract: The GP5 gene of porcine reproductive and respiratory syndrome virus (PRRSV) was integrated into the TK gene locus of pseudorabies virus (PRV) vaccine strain Bartha-K61, resulting in a TK- and gE- negative recombinant PRV harboring GP5 gene, designated as rPRV-GP5. The in vitro expression of the GP5 by rPRV-GP5-infected cells was analyzed by single-step growth analysis,Western blot,and indirect immunofluorescence test. It was shown that GP5 gene can be expressed authentically in the cytoplasm of rPRV-GP5-infected cells. Compared to its parental virus, rPRV-GP5 showed no obvious difference regarding viral replication and cytopathogenic effects in several cell cultures. Four PRV-negative sheep immunized intramuscularly with 10(6.0) PFU of rPRV-GP5 were fully protected from challenge with 10(3) LD50 of highly virulent PRV S strain of porcine origin. Ten PRV- and PRRSV-negative piglets given intranasally with 10(7.0) PFU of rPRV-GP5 and challenged intranasally with 10(5.0) TCID50 of virulent PRRSV CH-1a strain at day 63 post-inoculation developed antibodies against PRRSV 3, 5, 14 days post-challenge, as revealed by indirect immunofluorescence test, enzyme-linked immunosorbent assay and virus neutralization test. The results suggest that rPRV-GP5 is capable of inducing anamnestic immune response to PRRS in inoculated animals.
authors.affiliation: National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, CAAS, Harbin 150001, China.
authors.forename: Zhi-Jun
authors.initials: ZJ
authors.lastname: Tian
authors.forename: Hua-Ji
authors.initials: HJ
authors.lastname: Qiu
authors.forename: Jian-Qiang
authors.initials: JQ
authors.lastname: Ni
authors.forename: Yan-Jun
authors.initials: YJ
authors.lastname: Zhou
authors.forename: Xue-Hui
authors.initials: XH
authors.lastname: Cai
authors.forename: Guo-Hui
authors.initials: GH
authors.lastname: Zhou
authors.forename: Yun-Feng
authors.initials: YF
authors.lastname: Wang
authors.forename: Guang-Zhi
authors.initials: GZ
authors.lastname: Tong
mesh_terms: D000818:Animals; D005822:Genetic Vectors; D011558:Herpesvirus 1, Suid / Q000235:genetics*; D019316:Porcine respiratory and reproductive syndrome virus / Q000235:genetics / Q000502:physiology*; D011557:Pseudorabies; D014779:Virus Replication