Infectivity of Porcine Circovirus 1 and Circovirus 2 in Primary Porcine Hepatocyte and Kidney Cell Cultures

Hirai, Takuya; Nunoya, Tetsuo; Ihara, Takeshi; Saitoh, Toshiki; Shibuya, Kazumoto; Nakamura, Keigo

doi:10.1292/jvms.68.179

Cited by 11 publications

(6 citation statements)

References 17 publications

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“…In this study, a testis-derived epithelial-cell line (ST) was susceptible to PCV2 infection, showing that epithelial cells from other organs may also be susceptible to PCV2 infection. PCV2 infects epithelial cells in pigs experimentally infected with PCV2 (23), in PMWS pigs (13,57), and in primary kidney cultures (22), based on histological and morphological identification. This study confirmed the susceptibility of primary kidney epithelial cells to PCV2 infection using double immunofluorescence labeling of PCV2 antigens and cytokeratin, an epithelial-cell-specific intermediate filament.…”

Section: Discussionmentioning

confidence: 99%

Inhibition of Endosome-Lysosome System Acidification Enhances Porcine Circovirus 2 Infection of Porcine Epithelial Cells

Misinzo

Delputte

Nauwynck

2008

J Virol

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Section: Discussionmentioning

confidence: 99%

Inhibition of Endosome-Lysosome System Acidification Enhances Porcine Circovirus 2 Infection of Porcine Epithelial Cells

Misinzo

Delputte

Nauwynck

2008

J Virol

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“…A recent in vivo study reported that B cells may be the primary site of PCV2 replication during early infection (Yu et al, 2007), although in vitro studies have shown that PCV2 can replicate in primary porcine hepatocytes (Hirai et al, 2006) and that PCV2 particles can persist without replication in dendritic cells and porcine alveolar macrophages as well as in monocytes (Gilpin et al, 2003;Vincent et al, 2003). Thus, the target cells for PCV2 replication have not been clearly identified thus far.…”

Section: Discussionmentioning

confidence: 99%

Protective immunity against porcine circovirus 2 by vaccination with ORF2-based DNA and subunit vaccines in mice

Shen

Zhou

Huang

et al. 2008

Journal of General Virology

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The protective immune response against porcine circovirus 2 (PCV2) infection in mice was characterized using flow cytometric analysis (FCM), assays of antibody (of different IgG isotypes) and viraemia, and histopathological examination. An open reading frame 2 plasmid (pORF2) and the capsid protein (Cap) of PCV2 were used as DNA and subunit vaccines, respectively. In FCM analysis, although pORF2 and Cap alone showed comparable efficacy in eliciting lymphoproliferative responses and Cap-specific CD4 + T cells, pORF2 was superior to the Cap protein in triggering CD8 + T cells. A virus neutralization assay showed that pORF2 evoked stronger recall virus-neutralizing (VN) antibody responses than the Cap protein on PCV2 challenge. Correspondingly, VN antibody kinetics coincided with those of Cap-specific IgG2a, but not with the kinetics of IgG and IgG1. Following virus challenge, real-time PCR and histopathological analysis confirmed that only low viral DNA loads and mild microscopic lesions appeared in pORF2-immunized mice. These findings indicate that CD8 + T cells and VN antibody responses correlating mainly with Cap-specific IgG2a play crucial roles in protecting against PCV2 infection, and that the protective immunity induced by the pORF2 plasmid is superior to that induced by the PCV2 Cap protein. INTRODUCTIONPost-weaning multisystemic wasting syndrome (PMWS), first recorded in pigs in Western Canada in 1991, has been described in pigs from many regions of the world (Allan et al., 1998b;Allan & Ellis, 2000;Clark, 1997;Wen et al., 2005;Zhou et al., 2006). PMWS affects pigs of 5-12 weeks of age, with a morbidity of 5-30 %; it is characterized by weight loss, dyspnoea and jaundice. Field and experimental evidence has revealed that PMWS-affected pigs may develop severe immunosuppression (Segales et al., 2004).Porcine circovirus 2 (PCV2) has been identified as the primary aetiological agent of PMWS (Allan et al., 1998a, b;Allan & Ellis, 2000;Ellis et al., 1998;Fenaux et al., 2002;Meehan et al., 1998). PCV2 is a non-enveloped, singlestranded, circular DNA virus with a diameter of 17 nm (Tischer et al., 1982). The PCV2 genome comprises 1767 or 1768 nt and is assumed to have 11 potential open reading frames (ORF1-11;Hamel et al., 1998;Zhou et al., 2006). Previous studies have demonstrated that ORF1, -2 and -3 encode a 35.7 kDa replication (Rep) protein involved in virus replication (Mankertz et al., 1998), a 27.8 kDa capsid (Cap) protein involved in PCV2 immunogenicity (Mahe et al., 2000; Nawagitgul et al., 2000;Truong et al., 2001;Zhou et al., 2005a) and a protein involved in PCV2-induced apoptosis , respectively.Immunization against PCV2 has been studied intensely and found to be the most effective strategy for protecting pigs against PCV2 infection. Based on comparison of the immunogenicity of the PCV2 Cap and Rep proteins, Blanchard et al. (2003) demonstrated that the ORF2-encoded Cap protein was the major immunogen, whilst the ORF1-encoded Rep protein was only weakly immunogenic. Kamstrup et al. (2004) subsequently...

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“…The presence of C. perfringens harboring netB gene in tissue sections was detected by ISH. The ISH was performed as described previously [ 6 ] with some modifications. Briefly, deparaffinized sections were treated with 0.2 N HCl at room temperature for 20 min and then digested with proteinase K (Sigma-Aldrich Japan Inc., Tokyo, Japan) at 37°C for 15 min.…”

Section: Methodsmentioning

confidence: 99%

“…The sections were counterstained with Nuclear Fast Red (Roche Diagnostics GmbH) and systematically viewed under a light microscope. No background hybridization was seen when replicate tissue sections were incubated with an unrelated digoxigenin-labeled probe (PCV2-specific probe) [ 6 ] or when matched tissues from unaffected chickens were incubated with a netB gene-specific probe.…”

Section: Methodsmentioning

confidence: 99%

Experimental induction of necrotic enteritis in chickens by a <i>netB</i>-positive Japanese isolate of <i>Clostridium perfringens</i>

Suzuki

Kawahara

et al. 2017

The Journal of Veterinary Medical Science

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Necrotic enteritis (NE) is one of the most important bacterial diseases in terms of economic losses. Clostridium perfringens necrotic enteritis toxin B, NetB, was recently proposed as a new key virulent factor for the development of NE. The goal of this work was to develop a necrotic enteritis model in chickens by using a Japanese isolate of C. perfringens. The Japanese isolate has been found to contain netB gene, which had the same nucleotide and deduced amino acid sequences as those of prototype gene characterized in Australian strain EHE-NE18, and also expressed in vitro a 33-kDa protein identified as NetB toxin by nano-scale liquid chromatographic tandem mass spectrometry. In the challenge experiment, broiler chickens fed a commercial chicken starter diet for 14 days post-hatch were changed to a high protein feed mixed 50:50 with fishmeal for 6 days. At day 21 of age, feed was withheld for 24 hr, and each chicken was orally challenged twice daily with 2 ml each of C. perfringens culture (109 to 1010 CFU) on 5 consecutive days. The gross necrotic lesions were observed in 90 and 12.5% of challenged and control chickens, respectively. To our knowledge, this is the first study that demonstrated that a netB-positive Japanese isolate of C. perfringens is able to induce the clinical signs and lesions characteristic of NE in the experimental model, which may be useful for evaluating the pathogenicity of field isolates, the efficacy of a vaccine or a specific drug against NE.

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Infectivity of Porcine Circovirus 1 and Circovirus 2 in Primary Porcine Hepatocyte and Kidney Cell Cultures

Cited by 11 publications

References 17 publications

Inhibition of Endosome-Lysosome System Acidification Enhances Porcine Circovirus 2 Infection of Porcine Epithelial Cells

Inhibition of Endosome-Lysosome System Acidification Enhances Porcine Circovirus 2 Infection of Porcine Epithelial Cells

Protective immunity against porcine circovirus 2 by vaccination with ORF2-based DNA and subunit vaccines in mice

Experimental induction of necrotic enteritis in chickens by a <i>netB</i>-positive Japanese isolate of <i>Clostridium perfringens</i>

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