Identification of porcine alveolar macrophage glycoproteins involved in infection of porcine respiratory and reproductive syndrome virus

Wissink, E. H. J.; Wijk, Erwin van; Pol, J.M.A.; Godeke, Gert-Jan; Rijn, Piet A. van; Rottier, Peter J. M.; Meulenberg, J.J.M.

doi:10.1007/s00705-002-0897-0

Cited by 18 publications

(15 citation statements)

References 21 publications

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“…Masking epitopes in the GP5 5Ј total ectodomain, which have been hypothesized to account for a delayed appearance of neutralizing antibodies to PRRSV, do not explain our results, since a neutralizing function was not required for the detection of anti-GP5 antibody responses (51). Neutralizing antibodies are believed to be directed to conserved sequences in the GP5 ectodomain (24,25,27,28,43,52,53,55,61,62,65,66). Our results indicate that anti-GP5 antibodies do not play a significant role in the resolution of viremia and suggest that their delayed appearance contributes to the establishment of persistent infection.…”

Section: Vol 82 2008 B-cell Response To Prrsv 365mentioning

confidence: 76%

Antigen-Specific B-Cell Responses to Porcine Reproductive and Respiratory Syndrome Virus Infection

Mulupuri

Zimmerman

Hermann

et al. 2008

J Virol

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Porcine reproductive and respiratory syndrome virus (PRRSV) causes an acute, viremic infection of 4 to 6 weeks, followed by a persistent infection lasting for several months. We characterized antibody and B-cell responses to viral proteins in acute and persistent infection to better understand the immunological basis of the prolonged infection. The humoral immune response to PRRSV was robust overall and varied among individual viral proteins, with the important exception of a delayed and relatively weak response to envelope glycoprotein 5 (GP5). Memory B cells were in secondary lymphoid organs, not in bone marrow or Peyer's patches, in contrast to the case for many mammalian species. Potent anti-PRRSV memory responses were elicited to recall antigen in vitro, even though a second infection did not increase the B-cell response in vivo, suggesting that productive reinfection does not occur in vivo. Antibody titers to several viral proteins decline over time, even though abundant antigen is known to be present in lymphoid tissues, possibly indicating ineffective antigen presentation. The appearance of antibodies to GP5 is delayed relative to the resolution of viremia, suggesting that anti-GP5 antibodies are not crucial for resolving viremia. Lastly, viral infection had no immunosuppressive effect on the humoral response to a second, unrelated antigen. Taking these data together, the active effector and memory B-cell responses to PRRSV are robust, and over time the humoral immune response to PRRSV is effective. However, the delayed response against GP5 early in infection may contribute to the prolonged acute infection and the establishment of persistence.

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Section: Vol 82 2008 B-cell Response To Prrsv 365mentioning

confidence: 76%

Antigen-Specific B-Cell Responses to Porcine Reproductive and Respiratory Syndrome Virus Infection

Mulupuri

Zimmerman

Hermann

et al. 2008

J Virol

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“…Specifically, previous studies demonstrated that the minor envelope glycoproteins GP2 and GP4 of PRRSV interacted with pCD163 (51,52). The glycosylation of the GP2 and GP4 proteins was also shown to be required for efficient PRRSV interaction with the receptor (53,54). However, later studies demonstrated that only PRRSV GP4 protein interacted with pCD163 (55), and the glycosylation of GP4 might not play a vital role in this interaction (55,56).…”

Section: Pcd163 Srcr5 Structure For Prrsv Infectionmentioning

confidence: 99%

The Crystal Structure of the Fifth Scavenger Receptor Cysteine-Rich Domain of Porcine CD163 Reveals an Important Residue Involved in Porcine Reproductive and Respiratory Syndrome Virus Infection

Jiang

Qiao

et al. 2017

J Virol

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Porcine reproductive and respiratory syndrome (PRRS) has become an economically critical factor in swine industry since its worldwide spread in the 1990s. Infection by its causative agent, PRRS virus (PRRSV), was proven to be mediated by an indispensable receptor, porcine CD163 (pCD163), and the fifth scavenger receptor cysteine-rich domain (SRCR5) is essential for virus infection. However, the structural details and specific residues of pCD163 SRCR5 involved in infection have not been defined yet. In this study, we prepared recombinant pCD163 SRCR5 in Drosophila melanogaster Schneider 2 (S2) cells and determined its crystal structure at a high resolution of 2.0 Å. This structure includes a markedly long loop region and shows a special electrostatic potential, and these are significantly different from those of other members of the scavenger receptor cysteine-rich superfamily (SRCR-SF). Subsequently, we carried out structure-based mutational studies to identify that the arginine residue at position 561 (Arg561) in the long loop region is important for PRRSV infection. Further, we showed Arg561 probably takes effect on the binding of pCD163 to PRRSV during virus invasion. Altogether the current work provides the first view of the CD163 SRCR domain, expands our knowledge of the invasion mechanism of PRRSV, and supports a molecular basis for prevention and control of the virus.IMPORTANCE PRRS has caused huge economic losses to pig farming. The syndrome is caused by PRRSV, and PRRSV infection has been shown to be mediated by host cell surface receptors. One of them, pCD163, is especially indispensable, and its SRCR5 domain has been further demonstrated to play a significant role in virus infection. However, its structural details and the residues involved in infection are unknown. In this study, we determined the crystal structure of pCD163 SRCR5 and then carried out site-directed mutational studies based on the crystal structure to elucidate which residue is important. Our work not only provides structural information on the CD163 SRCR domain for the first time but also indicates the molecular mechanism of PRRSV infection and lays a foundation for future applications in prevention and control of PRRS.KEYWORDS CD163, Drosophila S2 cells, PRRSV, SRCR5, crystal structure P orcine reproductive and respiratory syndrome (PRRS) was first reported in 1987 in the United States (1) and has become a critical economic factor in the swine industry since its worldwide spread in the 1990s (2, 3). PRRS is characterized by

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“…Monoclonal antibodies against macrophage cell surface N-glycosylated proteins (220 kDa and 150 kDa doublet) block or reduce infection of macrophages [48]. A monoclonal antibody against a cytoskeletal filament complex (vimentin, cytokeratin 8, cytokeratin 18, actin, and hair type II basic keratin) blocks PRRSV infection of Marc-145 cells [25], and CD151, a member of the tetraspanin superfamily, also plays a critical role in PRRSV infection of Marc-145 cells [38].…”

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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Identification of porcine alveolar macrophage glycoproteins involved in infection of porcine respiratory and reproductive syndrome virus

Cited by 18 publications

References 21 publications

Antigen-Specific B-Cell Responses to Porcine Reproductive and Respiratory Syndrome Virus Infection

Antigen-Specific B-Cell Responses to Porcine Reproductive and Respiratory Syndrome Virus Infection

The Crystal Structure of the Fifth Scavenger Receptor Cysteine-Rich Domain of Porcine CD163 Reveals an Important Residue Involved in Porcine Reproductive and Respiratory Syndrome Virus Infection

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

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