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      Generation and Immunogenicity of a Recombinant Pseudorabies Virus Co-Expressing Classical Swine Fever Virus E2 Protein and Porcine Circovirus Type 2 Capsid Protein Based on Fosmid Library Platform

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          Abstract

          Pseudorabies (PR), classical swine fever (CSF), and porcine circovirus type 2 (PCV2)-associated disease (PCVAD) are economically important infectious diseases of pigs. Co-infections of these diseases often occur in the field, posing significant threat to the swine industry worldwide. gE/gI/TK-gene-deleted vaccines are safe and capable of providing full protection against PR. Classical swine fever virus (CSFV) E2 glycoprotein is mainly used in the development of CSF vaccines. PCV2 capsid (Cap) protein is the major antigen targeted for developing PCV2 subunit vaccines. Multivalent vaccines, and especially virus-vectored vaccines expressing foreign proteins, are attractive strategies to fight co-infections for various swine diseases. The gene-deleted pseudorabies virus (PRV) can be used to develop promising and economical multivalent live virus-vectored vaccines. Herein, we constructed a gE/gI/TK-gene-deleted PRV co-expressing E2 of CSFV and Cap of PCV2 by fosmid library platform established for PRV, and the expression of E2 and Cap proteins was confirmed using immunofluorescence assay and western blotting. The recombinant virus propagated in porcine kidney 15 (PK-15) cells for 20 passages was genetically stable. The evaluation results in rabbits and pigs demonstrate that rPRVTJ-delgE/gI/TK-E2-Cap elicited detectable anti-PRV antibodies, but not anti-PCV2 or anti-CSFV antibodies. These findings provide insights that rPRVTJ-delgE/gI/TK-E2-Cap needs to be optimally engineered as a promising trivalent vaccine candidate against PRV, PCV2 and CSFV co-infections in future.

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          Most cited references 64

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          Molecular biology of pseudorabies virus: impact on neurovirology and veterinary medicine.

          Pseudorabies virus (PRV) is a herpesvirus of swine, a member of the Alphaherpesvirinae subfamily, and the etiological agent of Aujeszky's disease. This review describes the contributions of PRV research to herpesvirus biology, neurobiology, and viral pathogenesis by focusing on (i) the molecular biology of PRV, (ii) model systems to study PRV pathogenesis and neurovirulence, (iii) PRV transsynaptic tracing of neuronal circuits, and (iv) veterinary aspects of pseudorabies disease. The structure of the enveloped infectious particle, the content of the viral DNA genome, and a step-by-step overview of the viral replication cycle are presented. PRV infection is initiated by binding to cellular receptors to allow penetration into the cell. After reaching the nucleus, the viral genome directs a regulated gene expression cascade that culminates with viral DNA replication and production of new virion constituents. Finally, progeny virions self-assemble and exit the host cells. Animal models and neuronal culture systems developed for the study of PRV pathogenesis and neurovirulence are discussed. PRV serves asa self-perpetuating transsynaptic tracer of neuronal circuitry, and we detail the original studies of PRV circuitry mapping, the biology underlying this application, and the development of the next generation of tracer viruses. The basic veterinary aspects of pseudorabies management and disease in swine are discussed. PRV infection progresses from acute infection of the respiratory epithelium to latent infection in the peripheral nervous system. Sporadic reactivation from latency can transmit PRV to new hosts. The successful management of PRV disease has relied on vaccination, prevention, and testing.
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            Porcine circovirus type 2 associated disease: update on current terminology, clinical manifestations, pathogenesis, diagnosis, and intervention strategies.

            Porcine circovirus type 2 (PCV2)-associated disease (PCVAD) continues to be an important differential diagnosis on pig farms in the United States and worldwide. Case trend analyses indicate that the incidence of PCVAD is on the rise in the United States. Accurate diagnosis is important in order to implement appropriate intervention strategies. PCVAD can manifest as a systemic disease, as part of the respiratory disease complex, as an enteric disease, as porcine dermatitis and nephropathy syndrome, or as reproductive problems. PCVAD may be only a sporadic individual animal diagnosis; however, PCVAD may also manifest as a severe herd problem accelerated and enhanced by concurrent virus or bacterial infections. This article is intended to discuss the most common disease manifestations, pathogenesis, diagnostic approaches, and intervention strategies associated with PCVAD in North America.
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              Experimental reproduction of severe wasting disease by co-infection of pigs with porcine circovirus and porcine parvovirus.

              Colostrum-deprived pigs were infected intranasally with a recent isolate of porcine circovirus (PCV2) and a porcine parvovirus (PPV), both from Canadian pigs with post-weaning multisystemic wasting syndrome (PMWS). Four pigs were inoculated with PCV2 alone, three with PPV alone, five with a combined PCV2/PPV inoculum, and two with a chloroform-treated combined PCV2/PPV inoculum. Pigs were killed 21-26 days after infection and tissue samples examined for gross and microscopical lesions and for the presence of viral antigens. No clinical signs, lesions or viral antigens were detected in two uninfected control pigs or in pigs inoculated with PPV alone. One pig inoculated with PCV2 alone became dull and thin. Mild to moderate histopathological lesions containing PCV2 antigen were detected in lymphoid tissues from the pigs inoculated with PCV2 alone. Pigs given the PCV2/PPV inoculum and the chloroform-treated PCV2/PPV inoculum became dull and two died. Jaundice and hepatomegaly were seen at post-mortem examination of most of the dually infected pigs. The latter showed large amounts of PCV2 antigen in numerous tissues; PPV antigen, which was less abundant, was detected in a few tissues, especially kidney. The lesions were similar to those seen in recently described field cases of porcine PMWS in North America and Europe. Copyright 1999 W.B. Saunders Company Ltd.
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                Author and article information

                Journal
                Pathogens
                Pathogens
                pathogens
                Pathogens
                MDPI
                2076-0817
                01 December 2019
                December 2019
                : 8
                : 4
                Affiliations
                State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150069, China; 2017y90100057@ 123456caas.cn (M.A.); 2017y90100180@ 123456caas.cn (T.T.); liyongfeng@ 123456caas.cn (Y.L.); wuhongxia@ 123456caas.cn (H.W.); wangtao02@ 123456caas.cn (T.W.)
                Author notes
                [* ]Correspondence: qiuhuaji@ 123456caas.cn (H.-J.Q.); sunyuan@ 123456caas.cn (Y.S.); Tel.: +86-451-5105-1708
                Article
                pathogens-08-00279
                10.3390/pathogens8040279
                6963705
                31805703
                © 2019 by the authors.

                Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ( http://creativecommons.org/licenses/by/4.0/).

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