Enhanced replication of porcine reproductive and respiratory syndrome (PRRS) virus in a homogeneous subpopulation of MA-104 cell line

Kim, H. S.; Kwang, Jimmy; Yoon, In Joong; Joo, Heesoo; Frey, M. L.

doi:10.1007/bf01313785

Cited by 573 publications

(367 citation statements)

References 12 publications

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“…The complete virus genomic sequence has been published (GenBank accession number PRU87392). Virus propagation and microinfectivity assays were performed on MARC-145 cells (National Veterinary Service Laboratory, Ames, IA, USA), a clone of the African monkey kidney cell line MA-104 that is considered highly permissive for PRRSV [23].…”

Section: Virus and Cellsmentioning

confidence: 99%

Effect of temperature and relative humidity on the stability of infectious porcine reproductive and respiratory syndrome virus in aerosols

et al. 2007

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-The objective of this experiment was to describe the stability of airborne infectious porcine reproductive and respiratory syndrome virus (PRRSV) as a function of temperature and relative humidity. A cloud of infectious PRRSV was aerosolized using 24-jet Collison nebulizer into a dynamic aerosol toroid (DAT) maintained at a specific temperature and relative humidity. The PRRSV cloud within the DAT was sampled repeatedly over time using SKC BioSampler impingers and the total viral RNA (RT-PCR) and concentration of infectious PRRSV (TCID 50 ) in the air samples was determined. As measured by quantitative RT-PCR, PRRSV RNA was stable under the conditions evaluated in this study. Thus, a comparison of viral RNA and Rhodamine B dye, a physical tracer, found no significant difference in the slopes of the lines. Titers of infectious virus were plotted by time and the half-life (T 1/2) of infectious PRRSV was calculated using linear regression analysis. An analysis of the results showed that aerosolized PRRSV was more stable at lower temperatures and/or lower relative humidity, but temperature had a greater effect on the T 1/2 of PRRSV than relative humidity. Based on these results, an equation was derived to predict the T 1/2 of infectious airborne PRRSV for any combination of environmental temperature and relative humidity.aerosol / virus stability / PRRSV / relative humidity / temperature

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Section: Virus and Cellsmentioning

confidence: 99%

Effect of temperature and relative humidity on the stability of infectious porcine reproductive and respiratory syndrome virus in aerosols

et al. 2007

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“…In vivo, PRRSV has a predilection for differentiated macrophages [18,44]. In vitro, primary porcine macrophages such as alveolar macrophages and the non-macrophage African green monkey kidney cell line MA-104, and cells derived from MA-104 (Marc-145 and CL-2621) support PRRSV infection [5,24,40,46,47]. Two PRRSV receptors on primary macrophages have already been identified.…”

Section: Introductionmentioning

confidence: 99%

Involvement of proteases in porcine reproductive and respiratory syndrome virus uncoating upon internalization in primary macrophages

2008

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-Porcine reproductive and respiratory syndrome virus (PRRSV) replicates in differentiated macrophages. In macrophages, heparan sulphate glycosaminoglycans mediate the initial PRRSV attachment and the receptor sialoadhesin mediates both PRRSV attachment and internalization into endosomes. Upon a pH drop, PRRSV is uncoated and its genome is released from the endosomes into the cytoplasm, which allows virus replication. However, expression of heparan sulphate and sialoadhesin in non-susceptible cells only allows virus internalization, but no virus uncoating and infection, indicating that other factors are involved. In the present study, it is shown that treatment of macrophages with serum (mainly the alpha-globulin fraction) inhibited PRRSV infection without affecting attachment and internalization. Because alpha-globulins contain several protease inhibitors, macrophages were treated with different protease inhibitors to investigate the involvement of proteases in PRRSV uncoating. Treatment of macrophages with broadly active inhibitors of serine or aspartic proteases, but not cysteine-or metallo-proteases, inhibited PRRSV uncoating and infection. Further investigation using specific inhibitors indicated that the aspartic protease cathepsin E is involved during PRRSV uncoating, but did not allow identification of the serine protease involved. The involvement of cathepsin E during PRRSV uncoating was confirmed by partial co-localization of internalized PRRSV with cathepsin E. Furthermore, cathepsin E expression increased with macrophage cultivation, which was positively correlated with an increased susceptibility to PRRSV infection. Together, these data show that, in macrophages, both the aspartic protease cathepsin E and an unidentified trypsin-like serine protease are involved in uncoating of internalized PRRSV and subsequent infection. porcine arterivirus / macrophage / virus entry / protease / receptor

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“…9 The first US PRRSV isolate was isolated on a continuous cell line, ATCC CL2621. 3 In addition to PAM cultures 9 and the CL2621 cell line, 3 the MARC 145 7 and ATCC CRL11171 8 cell lines also support PRRSV replication. Both US and European PRRSV isolates replicate well in PAM cultures, but the difficulties and expense in obtaining these cells limit their use.…”

mentioning

confidence: 99%

“…2,6 The variation in PRRSV propagation on various cell types has been reported, but differences due to medium or medium supplement changes have not been reported. 1,7 In this study, the growth of PRRSV isolates in CRL11171 a cell cultures maintained with either FBS or a serum replacement b was compared.To compare the replication of PRRSV with both supplements, CRL11171 cells were split into 2 populations, 1 maintained in Dulbecco's modified Eagle medium (DMEM) containing 10% FBS and the other maintained in DMEM supplemented with 10% serum replacement. After 3 passages in the respective supplemented medium, confluent monolayers of both cell populations were grown on 12-well plates.…”

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confidence: 99%

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Comparison of Porcine Reproductive and Respiratory Syndrome Virus Growth in Media Supplemented with Fetal Bovine Serum or a Serum Replacement

2001

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Abstract. Commercially available serum replacements are often used in cell culture as a cheaper and less variable substitute for fetal bovine serum (FBS). The growth of porcine reproductive and respiratory syndrome virus (PRRSV) isolates in CRL11171 cells maintained in a medium supplemented with FBS was compared with virus propagation in the same cell line maintained in the same medium with a serum replacement. The PRRSV replicated significantly better when the cell culture medium was supplemented with FBS. The results of this study have implications for the use of serum replacement-supplemented medium for PRRSV diagnosis by virus isolation.Porcine reproductive and respiratory syndrome virus (PRRSV) was first isolated in Europe on primary porcine alveolar macrophages (PAM) and was designated Lelystad virus. 9 The first US PRRSV isolate was isolated on a continuous cell line, ATCC CL2621. 3 In addition to PAM cultures 9 and the CL2621 cell line, 3 the MARC 145 7 and ATCC CRL11171 8 cell lines also support PRRSV replication. Both US and European PRRSV isolates replicate well in PAM cultures, but the difficulties and expense in obtaining these cells limit their use. Therefore, a continuous cell line is most often used to isolate and propagate PRRSV.Virus isolation is a commonly used method for the detection of PRRSV infection in many diagnostic laboratories. Clinical and diagnostic laboratories that maintain cell cultures for virus isolation can be affected by the cost and availability of fetal bovine serum (FBS). 6 Because FBS carries the risk of contamination with bovine viruses, batch-to-batch variation, and increased expense, a serum replacement is desirable, particularly in clinical and diagnostic settings. Commercially available serum replacements are often used as a substitute for FBS.Previous studies of serum replacements have shown variable results in their ability to support the growth of cell cultures and to replicate virus. Thus, the adaptability of cell lines to serum replacements and the capability of the cell line to propagate virus must be examined on an individual basis. 2,6 The variation in PRRSV propagation on various cell types has been reported, but differences due to medium or medium supplement changes have not been reported. 1,7 In this study, the growth of PRRSV isolates in CRL11171 a cell cultures maintained with either FBS or a serum replacement b was compared.To compare the replication of PRRSV with both supplements, CRL11171 cells were split into 2 populations, 1 maintained in Dulbecco's modified Eagle medium (DMEM) containing 10% FBS and the other maintained in DMEM supplemented with 10% serum replacement. After 3 passages in the respective supplemented medium, confluent monolayers of both cell populations were grown on 12-well plates. The cells showed no difference in growth rates or morphology regardless of the serum supplement used (data not shown). The CRL11171 monolayers were infected in triplicate with the 11th passage of VR-2385, a well-characterized virulent US PRRSV isolate, at...

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Enhanced replication of porcine reproductive and respiratory syndrome (PRRS) virus in a homogeneous subpopulation of MA-104 cell line

Cited by 573 publications

References 12 publications

Effect of temperature and relative humidity on the stability of infectious porcine reproductive and respiratory syndrome virus in aerosols

Effect of temperature and relative humidity on the stability of infectious porcine reproductive and respiratory syndrome virus in aerosols

Involvement of proteases in porcine reproductive and respiratory syndrome virus uncoating upon internalization in primary macrophages

Comparison of Porcine Reproductive and Respiratory Syndrome Virus Growth in Media Supplemented with Fetal Bovine Serum or a Serum Replacement

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