Production and titration of African swine fever virus in porcine alveolar macrophages

Carrascosa, Ángel L.; Santarén, Juán F.; Viñuela, Eladio

doi:10.1016/0166-0934(82)90034-9

Cited by 137 publications

(89 citation statements)

References 12 publications

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“…Jurkat human leukemia T cell line was obtained from the ATCC and cultured in RPMI 1640 medium supplemented with 10% FBS. Porcine alveolar macrophages were obtained by bronchoalveolar lavage of pigs and cultured in DMEM supplemented with 10% homologous swine serum (19). All medium were supplemented with 2 mM L-glutamine, 100 U/ml gentamicin, and nonessential amino acids.…”

Section: Cell Culture Viruses and Reagentsmentioning

confidence: 99%

The Viral Protein A238L Inhibits TNF-α Expression through a CBP/p300 Transcriptional Coactivators Pathway

Granja

Nogal

Hurtado

et al. 2006

The Journal of Immunology

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African swine fever virus (ASFV) is able to inhibit TNF-α-induced gene expression through the synthesis of A238L protein. This was shown by the use of deletion mutants lacking the A238L gene from the Vero cell-adapted Ba71V ASFV strain and from the virulent isolate E70. To further analyze the molecular mechanism by which the viral gene controls TNF-α, we have used Jurkat cells stably transfected with the viral gene to identify the TNF-α regulatory elements involved in the induction of the gene after stimulation with PMA and calcium ionophore. We have thus identified the cAMP-responsive element and κ3 sites on the TNF-α promoter as the responsible of the gene activation, and demonstrate that A238L inhibits TNF-α expression through these DNA binding sites. This inhibition was partially reverted by overexpression of the transcriptional factors NF-AT, NF-κB, and c-Jun. Furthermore, we present evidence that A238L inhibits the activation of TNF-α by modulating NF-κB, NF-AT, and c-Jun trans activation through a mechanism that involves CREB binding protein/p300 function, because overexpression of these transcriptional coactivators recovers TNF-α promoter activity. In addition, we show that A238L is a nuclear protein that binds to the cyclic AMP-responsive element/κ3 complex, thus displacing the CREB binding protein/p300 coactivators. Taken together, these results establish a novel mechanism in the control of TNF-α gene expression by a viral protein that could represent an efficient strategy used by ASFV to evade the innate immune response.

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Section: Cell Culture Viruses and Reagentsmentioning

confidence: 99%

The Viral Protein A238L Inhibits TNF-α Expression through a CBP/p300 Transcriptional Coactivators Pathway

Granja

Nogal

Hurtado

et al. 2006

The Journal of Immunology

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“…A virulent ASF virus isolate (CC83) and a non-virulent one (BA71) grown in porcine alveolar macrophages were used. ASF virus titrations were performed in porcine macrophages as described elsewhere (Carrascosa et al, 1982). Titres were calculated by the method of Reed & Muench (1938) and expressed as haemadsorption units per ml (HADU/ml).…”

Section: Introductionmentioning

confidence: 99%

“…Cells. Porcine alveolar macrophages were collected by alveolar lavage as previously described (Carrascosa et aL, 1982) and kept frozen until use. Porcine peripheral blood mononuclear cells were isolated by sedimenting a mixture of heparinized blood and dextran (Sigma) followed by centrifugation in a Ficoll-Hypaque (Pharmacia) density gradient as described elsewhere (Boyum, 1968).…”

Section: Introductionmentioning

confidence: 99%

Effect of Interferon- , Interferon- and Tumour Necrosis Factor on African Swine Fever Virus Replication in Porcine Monocytes and Macrophages

Esparza

González

Viñuela

1988

Journal of General Virology

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SUMMARYBovine interferon-0ql (IFN-cti1) and porcine interferon-y (IFN-~) inhibited African swine fever virus replication in both porcine monocytes and alveolar macrophages. The most potent antiviral activity was observed with IFN-y-treated alveolar macrophages. The production of both a virulent (CC83) and a non-virulent (BA71) isolate of the virus was inhibited. Bovine turnout necrosis factor ~ did not show antiviral activity in either monocytes or alveolar macrophages. Rather, an increase of African swine fever virus production in tumour necrosis factor or-treated monocytes was found. An analysis of viral protein synthesis in IFN-~tI1-and IFN-~,-treated alveolar macrophages showed an inhibition of synthesis of some viral proteins. The inhibition of late proteins was very pronounced in IFN-~-treated cells, and it was probably a consequence of the inhibition of African swine fever virus DNA polymerase activity.

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“…On day 5, rpTNF-α was added to induce their maturation. Alveolar macrophages were collected by bronchoalveolar lavage as described by Carrascosa et al [9].…”

Section: Tissues and Cellsmentioning

confidence: 99%

Expression of toll-like receptor 2 (TLR2) in porcine leukocyte subsets and tissues

Álvarez

Revilla

Doménech³

et al. 2007

Vet. Res.

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-Toll-like receptors (TLR) are a group of pattern recognition molecules that play a crucial role in innate immunity. TLR2 recognises a variety of microbial components leading to the development of inflammatory and immune responses. To characterise the expression and functional properties of porcine TLR2 (pTLR2), we have raised a panel of monoclonal antibodies (mAb) against this molecule. Mouse 3T3 cell transfectants expressing pTLR2 were used for immunisation of mice. The specificity of these antibodies was confirmed by their reactivity with CHO cells transfected with pTLR2 but not with pTLR4 or with non-transfected cells. Using one of these mAbs, named 1H11, pTLR2 was found on cells of the innate immune system, including monocytes, macrophages, and granulocytes, but not on peripheral blood lymphocytes. Staining of tissue sections showed that pTLR2 is also expressed on epithelial cells lining the tracheobronchial and intestinal tracts, bile ducts in the liver and renal tubules, and on the basal layer of the epidermis. This distribution is consistent with a surveillance function at entry sites, allowing for early detection of microbial invasion.toll-like receptor / monoclonal antibody / monocyte / epithelial cell / pig

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Production and titration of African swine fever virus in porcine alveolar macrophages

Cited by 137 publications

References 12 publications

The Viral Protein A238L Inhibits TNF-α Expression through a CBP/p300 Transcriptional Coactivators Pathway

The Viral Protein A238L Inhibits TNF-α Expression through a CBP/p300 Transcriptional Coactivators Pathway

Effect of Interferon- , Interferon- and Tumour Necrosis Factor on African Swine Fever Virus Replication in Porcine Monocytes and Macrophages

Expression of toll-like receptor 2 (TLR2) in porcine leukocyte subsets and tissues

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