Isolation of Swine Infertility and Respiratory Syndrome Virus (Isolate ATCC VR-2332) in North America and Experimental Reproduction of the Disease in Gnotobiotic Pigs

Collins, James E.; Benfield, David A.; Christianson, William T.; Harris, Louis; Hennings, Jane C; Shaw, Daniel P.; Goyal, Sagar M.; McCullough, Sam; Morrison, Robert B.; Joo, Heesoo; Gorcyca, David E.; Chladek, Dan

doi:10.1177/104063879200400201

Cited by 707 publications

(447 citation statements)

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“…Now the disease occurs in most pig-producing countries and causes great economic loss to swine industry worldwide except for few countries including Switzerland, New Zealand and Australia [7,21]. The causative agent, PRRSV, is an enveloped, single-stranded, positive-sense RNA virus with an approximate diameter of 50-65 nm and belongs to the family Arteriviridae in the order of Nidovirales [8,31]. There are two distinct genotypes of PRRSV: genotype 1 (European) and genotype 2 (North American) which, share only about 60 % identity on the nucleotide level, while individual PRRSV isolates within genotypes varies approximately 20 % in nucleotide sequence [13,19].…”

Section: Introductionmentioning

confidence: 99%

Indian porcine reproductive and respiratory syndrome virus bears discontinuous deletion of 30 amino acids in nonstructural protein 2

et al. 2016

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Since its first outbreak in 2013, porcine reproductive and respiratory syndrome (PRRS) has established as an enzootic disease in pig population of Mizoram state, India. Our previous studies based on phylogenetic analysis of ORF5 and ORF7 gene sequences revealed close relationship of Indian PRRSV with the highly pathogenic variant of PRRSV (HP-PRRSV) of Chinese origin. Despite the control measures, second major outbreak of the disease was recorded in Aizawl district of Mizoram in 2015. The objective of the present study was to examine the origin of PRRSV isolates of 2015 outbreak, identification of deleted region in Nsp2 gene and determination of any genetic variation between 2013 and 2015 isolates of PRRSV. The outbreak was confirmed by the detection of PRRSVspecific antibodies in 57 out of 92 serum samples (61.96 %) and also by RT-PCR in 42 out of 42 necropsy samples (100 %). Nucleotide sequence analysis of Nsp2 coding region of Indian isolates and comparison with reference sequences revealed 90 nucleotides discontinuous deletion further establishes the closeness of Indian PRRSV to Chinese HP-PRRSV. Further, sequence and phylogenetic analysis of ORF5, ORF7 and Nsp2 genes of Indian PRRSV from both 2013 and 2015 revealed that the outbreaks were caused by two different strains of HP-PRRSV closely associated with the Chinese 10 HEB-3 isolate and 07QN isolates of Vietnam origin respectively. The present study confirms that the Indian PRRSV is a highly pathogenic variant of PRRSV and this study serves as the basis for developing practical and effective control measures against this disease.

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Section: Introductionmentioning

confidence: 99%

Indian porcine reproductive and respiratory syndrome virus bears discontinuous deletion of 30 amino acids in nonstructural protein 2

et al. 2016

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“…* Corresponding author: peter.delputte@UGent.be problems in pigs of all ages, in combination with other pathogens [7,8,21,28]. In vivo, PRRSV has a predilection for differentiated macrophages [18,44].…”

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.…”

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confidence: 99%

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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Isolation of Swine Infertility and Respiratory Syndrome Virus (Isolate ATCC VR-2332) in North America and Experimental Reproduction of the Disease in Gnotobiotic Pigs

Cited by 707 publications

References 10 publications

Indian porcine reproductive and respiratory syndrome virus bears discontinuous deletion of 30 amino acids in nonstructural protein 2

Indian porcine reproductive and respiratory syndrome virus bears discontinuous deletion of 30 amino acids in nonstructural protein 2

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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