Generation of a porcine alveolar macrophage cell line for the growth of porcine reproductive and respiratory syndrome virus

Lee, Yoo Jin; Park, Choi‐Kyu; Nam, Eeuri; Kim, Seong‐Hee; Lee, O-Soo; Lee, Du Sik; Lee, Changhee

doi:10.1016/j.jviromet.2009.11.003

Cited by 87 publications

(58 citation statements)

References 27 publications

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“…We have shown that infection of our PBMMWs results in a productive infection similar to that observed in alveolar macrophages and bone marrow-derived dendritic cells (Costers et al, 2008;Lee et al, 2010). Since we observed an earlier peak of virus production (24 h versus 36-48 h), we chose a 24 h time point to assay phagosomal function.…”

Section: Discussionmentioning

confidence: 90%

“…The nature of alveolar macrophage activation is determined not only by the genetic lineage of the macrophages, but by the environment in which they are derived and survive: they are constantly exposed to inhaled dust, pollutants and microbes and as such have to maintain a constant 'deactivated' phenotype (du Manoir et al, 2002;Lambrecht, 2006), and thus can be primed in different ways depending on the environment in which the animal was raised. The immortalized PAM model developed by Lee et al (2010) is an excellent model with which to study PRRSV. Culturing PAM in vitro removes the risk of differential priming of macrophages and genetic and environmental differences in the macrophages.…”

Section: Discussionmentioning

confidence: 99%

“…PAMs are also primary cells which do not divide, and their functionality is heavily dependent on the age of the animal and the environment in which the animal lives (du Manoir et al, 2002). Lee et al (2010) developed an easy-to-culture cell line derived from PAM that stably express CD163, and are thus susceptible to PRRSV infection. We set out to add to this model using a primary cell model that was relatively easy to harvest, culture, manipulate and characterize.…”

Section: L929-conditioned Medium Drives Differentiation Of Macrophagementioning

confidence: 99%

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Infection of porcine bone marrow-derived macrophages by porcine respiratory and reproductive syndrome virus impairs phagosomal maturation

Chaudhuri

McKenna

Balce

et al. 2016

Journal of General Virology

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Porcine reproductive and respiratory syndrome virus (PRRSV), a positive-sense, ssRNA virus of the genus Arterivirus, is a devastating disease of swine worldwide. Key early targets of PRRSV infection in pigs include professional phagocytes in the lung, such as alveolar and interstitial macrophages and dendritic cells, the dysfunction of which is believed to be responsible for much of the associated mortality. In order to study the effect of virus infection on phagocyte function, the development of a robust, reproducible model would be advantageous. Given the limitations of current models, we set out to develop a porcine bone marrow-derived macrophage (PBMMW) cell model to study phagosomal maturation and function during PRRSV infection. Derivation of PBMMWs from marrow using cultured L929 fibroblast supernatant produced a homogeneous population of cells that exhibited macrophage-like morphology and proficiency in Fc-receptor-mediated phagocytosis and phagosomal maturation. PBMMWs were permissive to PRRSV infection, resulting in a productive infection that peaked at 24 h. Assessment of the effect of PRRSV infection on the properties of phagosomal maturation in PBMMWs revealed a significant decrease in phagosomal proteolysis and lowered production of reactive oxygen species, but no change in PBMMW viability, phagocytosis or the ability of phagosomes to acidify. In this study, we present a new model to investigate PRRSV infection of phagocytes, which demonstrates a significant effect on phagosomal maturation with the associated implications on proper macrophage function. This model can also be used to study the effect on the phagosomal microenvironment of infection by other viruses targeting porcine macrophages.

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Section: Discussionmentioning

confidence: 90%

Section: Discussionmentioning

confidence: 99%

Section: L929-conditioned Medium Drives Differentiation Of Macrophagementioning

confidence: 99%

See 1 more Smart Citation

Infection of porcine bone marrow-derived macrophages by porcine respiratory and reproductive syndrome virus impairs phagosomal maturation

Chaudhuri

McKenna

Balce

et al. 2016

Journal of General Virology

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“…The PEE cell line (passage 15), as well as the primary cells and PAM, was seeded on 24-well plates (1 × 10 5 cells/well), and cultivated for 24 h. After fixation with 4% paraformaldehyde and blocking with PBSM (5% defatted milk powder in PBS, pH7.4), indirect immunofluorescence was performed as previously described (Lee et al, 2010) using anti-CD34, CD151, CD163 and Sn polyclonal antibodies. For detection of the viral antigen, cells were infected with PRRSV strain VR2332 (MOI 0.1) and submitted to immunofluorescence 24 HPI using the mAb against PRRSV N protein.…”

Section: Immunofluorescencementioning

confidence: 99%

Generation and characterization of a porcine endometrial endothelial cell line susceptible to porcine reproductive and respiratory syndrome virus

Zhang

Xia

et al. 2013

Virus Research

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“…The retrovirus gene transfer system (Stratagene, USA) was applied to generate BHK cell lines constitutively expressing the recombinant ORF7 gene [2]. The selected cell clones (BHK-EU-ORF7 and BHK-NA-ORF7) in the presence of 800 µg/ml G418 (Invitrogen, USA) were initially subjected to PCR to identify the PRRSV ORF7 gene integration, followed by RT-PCR and nucleotide sequencing to determine N gene expression at the mRNA level.…”

mentioning

confidence: 99%

Development and characterization of stable cell lines constitutively expressing the porcine reproductive and respiratory syndrome virus nucleocapsid protein

et al. 2010

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Despite global efforts to control porcine reproductive and respiratory syndrome virus (PRRSV) infection, the virus continues to cause economic problems in the swine industry worldwide. In this study, we attempted to generate and characterize a panel of stable BHK cell lines that constitutively express the nucleocapsid (N) protein of type 1 or type 2 PRRSV. The established BHK cell lines were found to react well with N-specific antibodies as well as the hyperimmune serum of pigs raised against each genotype of PRRSV. Taken together, the data implicate a potential usefulness for the newly generated stable cell lines as a diagnostic reagent for PRRSV serology.

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Generation of a porcine alveolar macrophage cell line for the growth of porcine reproductive and respiratory syndrome virus

Cited by 87 publications

References 27 publications

Infection of porcine bone marrow-derived macrophages by porcine respiratory and reproductive syndrome virus impairs phagosomal maturation

Infection of porcine bone marrow-derived macrophages by porcine respiratory and reproductive syndrome virus impairs phagosomal maturation

Generation and characterization of a porcine endometrial endothelial cell line susceptible to porcine reproductive and respiratory syndrome virus

Development and characterization of stable cell lines constitutively expressing the porcine reproductive and respiratory syndrome virus nucleocapsid protein

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