Production of a monoclonal antibody against Porcine circovirus type 3 cap protein

Li, Xiangdong; Bai, Yilin; Zhang, Haiyang; Zheng, Dingding; Wang, Tongyan; Wang, Yanwei; Deng, Junhua; Sun, Zhe; Tian, Kegong

doi:10.1016/j.jviromet.2018.07.014

Cited by 21 publications

(19 citation statements)

References 13 publications

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“…The similarity between PCV-3 sequences ranges from 97 to 100% throughout the analyzed years and tested countries (48, 69–71). Phylogenetic analyses suggested two main groups classified as PCV-3a and PCV-3b and several sub-clusters (48, 72, 73), based on differences found between both groups in the aa sites 122 and 320 (S122A and A320V). In fact, certain antigenicity differences among groups have been proposed (74), although it is still too early to discuss about potential different genotypes or subgroups for PCV-3.…”

Section: Molecular Organization Of Porcine Circovirusesmentioning

confidence: 99%

Current Knowledge on Porcine circovirus 3 (PCV-3): A Novel Virus With a Yet Unknown Impact on the Swine Industry

et al. 2018

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Porcine circovirus 3 (PCV-3) is a recently described virus belonging to the family Circoviridae. It represents the third member of genus Circovirus able to infect swine, together with PCV-1, considered non-pathogenic, and PCV-2, one of the most economically relevant viruses for the swine worldwide industry. PCV-3 was originally found by metagenomics analyses in 2015 in tissues of pigs suffering from porcine dermatitis and nephropathy syndrome, reproductive failure, myocarditis and multisystemic inflammation. The lack of other common pathogens as potential infectious agents of these conditions prompted the suspicion that PCV-3 might etiologically be involved in disease occurrence. Subsequently, viral genome was detected in apparently healthy pigs, and retrospective studies indicated that PCV-3 was already present in pigs by early 1990s. In fact, current evidence suggests that PCV-3 is a rather widespread virus worldwide. Recently, the virus DNA has also been found in wild boar, expanding the scope of infection susceptibility among the Suidae family; also, the potential reservoir role of this species for the domestic pig has been proposed. Phylogenetic studies with available PCV-3 partial and complete sequences from around the world have revealed high nucleotide identity (>96%), although two main groups and several subclusters have been described as well. Moreover, it has been proposed the existence of a most common ancestor dated around 50 years ago. Taking into account the economic importance and the well-known effects of PCV-2 on the swine industry, a new member of the same family like PCV-3 should not be neglected. Studies on epidemiology, pathogenesis, immunity and diagnosis are guaranteed in the next few years. Therefore, the present review will update the current knowledge and future trends of research on PCV-3.

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Section: Molecular Organization Of Porcine Circovirusesmentioning

confidence: 99%

Current Knowledge on Porcine circovirus 3 (PCV-3): A Novel Virus With a Yet Unknown Impact on the Swine Industry

et al. 2018

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“…Circoviruses are the smallest autonomously replicating DNA viruses, and they show a typical ambisense genomic structure [9,10]. Circoviruses have a non-enveloped, circular, single-stranded DNA genome consisting of two major open reading frames, ORF1 and ORF2, which code the replicase (Rep) and the capsid protein (Cap), respectively [11,12]. As in PCV1 and PCV2, PCV3 ORF2 encodes the only structural capsid protein, which contains the dominant immunological regions [11,13].…”

Section: Introductionmentioning

confidence: 99%

“…Circoviruses have a non-enveloped, circular, single-stranded DNA genome consisting of two major open reading frames, ORF1 and ORF2, which code the replicase (Rep) and the capsid protein (Cap), respectively [11,12]. As in PCV1 and PCV2, PCV3 ORF2 encodes the only structural capsid protein, which contains the dominant immunological regions [11,13]. Circoviruses lack an autonomous DNA polymerase and are dependent on the replication machinery of the host cell for de novo DNA synthesis.…”

Section: Introductionmentioning

confidence: 99%

Identification of Nuclear Localization Signals in the ORF2 Protein of Porcine Circovirus Type 3

Mou

Wang

Pan

et al. 2019

Viruses

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Porcine circovirus type 3 (PCV3) contains two major open reading frames (ORFs) and the ORF2 gene encodes the major structural capsid protein. In this study, nuclear localization of ORF2 was demonstrated by fluorescence observation and subcellular fractionation assays in ORF2-transfected PK-15 cells. The subcellular localization of truncated ORF2 indicated that the 38 N-terminal amino acids were responsible for the nuclear localization of ORF2. The truncated and site-directed mutagenesis of this domain were constructed, and the results demonstrated that the basic amino acid residues at positions 8-32 were essential for the strict nuclear localization. The basic motifs 8 RRR-R-RRR 16 and 16 RRRHRRR 22 were further shown to be the key functional nucleolar localization signals that guide PCV3 ORF2 into nucleoli. Furthermore, sequence analysis showed that the amino acids of PCV3 nuclear localization signals were highly conserved. Overall, this study provides insight into the biological and functional characteristics of the PCV3 ORF2 protein.case of PCVs, the N-terminus of ORF2 is rich in basic amino acids and displays nuclear localization signals (NLSs) [14][15][16]. The NLS is a short stretch of amino acids that mediates the transport of nuclear proteins into the nucleus. NLS motifs play a key role in this mechanism. NLS sequences are often composed of basic amino acids and can be classified as either monopartite or bipartite motifs [17,18]. The nuclear targeting of PCV2 ORF2 is directed by the bipartite motifs situated at the N-terminus of the proteins [14,15]. NLSs of PCV1 ORF2 show high homologies to classical monopartite or bipartite NLS, which is essential for the complete nuclear import of PCV1 ORF2. The N-terminus of PCV1 and PCV2 ORF2 contain several conserved basic amino acid stretches [15,19], and shares 70.7% in nucleotide identity and 82.9% in amino acid similarity.Verification of NLS and demonstration of intracellular distribution facilitate understanding of viral protein function. The amino acids of PCV3 ORF2 are markedly different from those of the other PCVs. This led us to study the functional motifs in nuclear targeting of PCV3 ORF2. In this study, a series of recombinant plasmids expressing PCV3 ORF2 fused to EGFP were constructed, and we identified the main motifs of the NLSs mediating nuclear localization of PCV3 ORF2.

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“…In our previous study, we identified a PCV3-specific monoclonal antibody, 1H11, which can distinguish PCV3 from other PCV genotypes specifically and accurately [29]. In the present study, we expressed a soluble form of PCV3 capsid protein with SUMO-tag using Escherichia coli expression system and successfully obtained in vitro assembled PCV3-VLPs.…”

Section: Introductionmentioning

confidence: 86%

“…Amongst them, the shorter CD-loop of PCV3 capsid protein contains much lower densities and significantly different structural conformations as compared with those of PCV2 capsid protein. Our previous reports show that PCV3-VLPs could produce PCV3-specific monoclonal antibodies in the host [29].…”

Section: Structural and Sequence Comparisons Of Pcv Capsid Proteinsmentioning

confidence: 97%

Structural insight into the type-specific epitope of porcine circovirus type 3

Li²,

Zhai

et al. 2020

Bioscience Reports

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The recently identified pathogenic Porcine circovirus type 3 (PCV3) may threaten to reduce the pig population dramatically worldwide. In our previous study, a PCV3-specific monoclonal antibody (mAb-1H11) was successfully applied in immune-histochemistry staining and ELISA, which specifically recognize PCV3 capsid protein in PCV3-positive pig tissues. In the present study, we expressed and purified the soluble sole capsid protein of PCV3. The purified capsid protein was capable of self-assembly into virus-like-particles (VLPs), which is validated by transmission electron microscopy and dynamic light scattering assays. Moreover, the epitope of mAb-1H11 was identified in the CD-loop region (a.a. 72-79) on the VLP surface, which is confirmed by PCV2-PCV3 epitope swapping assay. For the first time, we determined the cryo-EM structure of PCV3-VLP at 8.5 Å resolution that reveals the detailed structural information of PCV3-VLP. In our cryo-EM structure, PCV3-VLP is composed of 60 capsid protein subunits assembled with T = 1 icosahedral symmetry. Consistent to our bio-dot Western blot assay, the structural comparison between PCV3 and PCV2 revealed significant structural differences in the surface-exposed loops, including the CD-loop (a.a. 72-79) and the EF-loop (a.a. 109-131). Our work provides a structural framework for engineering future PCV3 vaccine and diagnosis kits development.

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Production of a monoclonal antibody against Porcine circovirus type 3 cap protein

Cited by 21 publications

References 13 publications

Current Knowledge on Porcine circovirus 3 (PCV-3): A Novel Virus With a Yet Unknown Impact on the Swine Industry

Current Knowledge on Porcine circovirus 3 (PCV-3): A Novel Virus With a Yet Unknown Impact on the Swine Industry

Identification of Nuclear Localization Signals in the ORF2 Protein of Porcine Circovirus Type 3

Structural insight into the type-specific epitope of porcine circovirus type 3

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