Seroneutralization of Porcine Reproductive and Respiratory Syndrome Virus Correlates with Antibody Response to the GP5Major Envelope Glycoprotein

Abstract Immunoglobulin G antibody response, measured as sample-to-positive (S/P) ratio, to Porcine Reproductive and Respiratory Syndrome virus (PRRSV) has been proposed as an indicator trait for improved reproductive performance in PRRSV-infected purebred sows and PRRSV-vaccinated crossbred gilts. In this study, we investigated the genetic correlations (rg) of S/P ratio following a PRRSV outbreak and PRRSV-vaccination with performance in non-exposed and PRRSV-exposed sows. PRRSV outbreak phase was defined based on previously described methodologies after the detection of typical clinical signs of PRRSV infection. 541 Landrace sows had S/P ratio measured at ~54 days after the beginning of the PRRSV outbreak (S/Poutbreak), and 906 Landrace x Large White naïve F1 gilts had S/P ratio measured at ~50 days after vaccination with a commercial modified live PRRSV vaccine (S/PVx). 711 and 428 Landrace sows had reproductive performance recorded before and during the PRRSV outbreak, respectively. 811 vaccinated F1 animals had farrowing performance for up to 3 parities. All animals were genotyped for ~28K SNPs. The estimate of rg of S/Poutbreakwith S/PVx was high (rg±SE = 0.72±0.18). Estimates of rg of S/Poutbreak with reproductive performance in F1 sows were low to moderate, ranging from 0.05±0.23 (number stillborn) to 0.30±0.20 (total number born). Estimates of rg of S/PVxwith reproductive performance in non-infected purebred sows were moderate and favorable with number born alive (0.50±0.23), but low (0 to -0.11±0.23) with litter mortality traits. Estimates of rg of S/PVx were moderate and negative (-0.47±0.18) with the number of mummies in PRRSV-infected purebred sows and low with other traits (-0.29±0.18 for total number born to 0.05±0.18 for number stillborn). These results indicate that selection for antibody response following a PRRSV outbreak collected in purebred sows and to PRRSV vaccination collected in commercial crossbred gilts may increase litter size of non-infected and PRRSV-exposed purebred and commercial crossbred sows.

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Fine Analysis of Humoral Antibody Response to Envelope Glycoprotein of SIV in Infected and Vaccinated Macaques

AIDS Research and Human Retroviruses ◽

10.1089/aid.1994.10.1295 ◽

1994 ◽

Vol 10 (10) ◽

pp. 1295-1304 ◽

Cited By ~ 16

Author(s):

P. SILVERA ◽

B. FLANAGAN ◽

K. KENT ◽

E. RUD ◽

C. POWELL ◽

...

Keyword(s):

Antibody Response ◽

Envelope Glycoprotein ◽

Humoral Antibody

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Identification of an immunodominant epitope in the C terminus of glycoprotein 5 of porcine reproductive and respiratory syndrome virus

Journal of General Virology ◽

10.1099/0022-1317-82-5-995 ◽

2001 ◽

Vol 82 (5) ◽

pp. 995-999 ◽

Cited By ~ 26

Author(s):

María Jose Rodriguez ◽

Javier Sarraseca ◽

Jesús Fominaya ◽

Elena Cortés ◽

Antonio Sanz ◽

...

Keyword(s):

Antibody Response ◽

Virus Neutralization ◽

Respiratory Syndrome Virus ◽

Deletion Mutants ◽

Immunodominant Epitope ◽

Diagnostic Potential ◽

C Terminus ◽

Transmembrane Regions ◽

Glycoprotein 5

Glycoprotein 5 (GP5) is the major glycoprotein of porcine reproductive and respiratory syndrome virus (PRRSV). Expression of GP5 has been improved by removing the transmembrane regions. Vectors were constructed encoding complete GP5 plus three mutants: GP5 ΔNs (residues 28–201), GP5[30–67] (residues 30–67) and GP5[30–201] (residues 30–67/130–201). The three deletion mutants were expressed at levels 20–30 times higher than complete GP5. GP5[30–201] was well recognized in ELISA or immunoblotting by a collection of pig sera. All the fragments were tested for the generation of MAbs, but only the polyhistidine-tagged fragment GP5[30–201]H elicited an antibody response sufficient to produce MAbs. The two MAbs were positive for PRRSV in ELISA and immunoblotting, but negative for virus neutralization. MAb 4BE12 reacted with residues 130–170 and MAb 3AH9 recognized residues 170–201. This region was recognized strongly in immunoblotting by a collection of infected-pig sera. These results indicate diagnostic potential for this epitope.

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Induction of auto-anti-idiotypic antibodies specific for antibodies to matrix and envelope glycoprotein from pigs experimentally infected with porcine reproductive and respiratory syndrome virus

Veterinary Immunology and Immunopathology ◽

10.1016/j.vetimm.2004.03.007 ◽

2004 ◽

Vol 101 (1-2) ◽

pp. 49-59 ◽

Cited By ~ 6

Author(s):

En-Min Zhou ◽

Alfonso Clavijo ◽

Zhengjun Jiang ◽

Mehrdad Ameri-Mahabadi ◽

Jeffrey J Zimmerman

Keyword(s):

Envelope Glycoprotein ◽

Respiratory Syndrome Virus

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The administration of diets contaminated with low to intermediate doses of deoxynivalenol and supplemented with antioxidants and binding agents slightly affects the growth, antioxidant status and vaccine response in weanling pigs

Journal of Animal Science ◽

10.1093/jas/skab238 ◽

2021 ◽

Author(s):

Luca Lo Verso ◽

Kristina Dumont ◽

Martin Lessard ◽

Karoline Lauzon ◽

Chantale Provost ◽

...

Keyword(s):

Antibody Response ◽

Antioxidant Status ◽

Block Design ◽

Basal Diet ◽

Respiratory Syndrome Virus ◽

Vaccine Response ◽

Weanling Pigs ◽

Tocopherol Concentration ◽

Reducing Ability ◽

The Impact

Abstract This study aimed to evaluate the impact of grading levels of deoxynivalenol (DON) in the diet of weaned pigs, as well as the effects of a supplementation with antioxidants (AOX), hydrated sodium calcium aluminosilicates (HSCAS) and their combination on the growth, antioxidant status, immune and vaccine response against the porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circovirus 2 (PCV2). At weaning, 336 piglets were allocated to six dietary treatments according to a randomized complete block design. Treatments were as follows: basal diet (CTRL); basal diet containing DON at 1.2 mg/kg (DON1.2); basal diet containing DON at 2.4 mg/kg (DON2.4); DON2.4 diet + a mix of AOX which included vitamins A and E at 20,000 IU and 200 IU/kg feed respectively, selenized yeast at 0.3 mg/kg and a grape seed extracts at 100 mg/kg feed (DON2.4+AOX); DON2.4 diet + modified HSCAS at 1 g/kg (DON2.4+HSCAS); DON2.4+AOX+HSCAS. Pigs were vaccinated against PRRSV and PCV2 at 7 d; at 0, 14 and 35 d growth performance were recorded, and blood samples were collected in order to evaluate the oxidative status, inflammatory blood markers, lymphocyte blastogenic response and vaccine antibody response. Increasing intake of DON resulted in a quadratic effect at 35 d in the lymphocyte proliferative response to Concanavalin A and PCV2 as well as in the anti-PRRSV antibody response, whereas the catalase activity decreased in DON2.4 pigs compared to the CTRL and DON1.2 groups (P ≤ 0.05). Compared to the DON2.4 diet, the AOX supplementation slightly reduced G:F ratio (P = 0.026) and increased the ferric reducing ability of plasma as well as α-tocopherol concentration (P < 0.05), whereas the association AOX+HSCAS increased the anti-PRRSV IgG (P < 0.05). Furthermore, the HSCAS supplement reduced haptoglobin levels in serum at 14 d compared to the DON2.4 group; however, its concentration decreased in all the experimental treatments from 14 d to 35 d and particularly in the DON2.4+AOX pigs, whereas a different trend was evidenced in the DON2.4+HSCAS group, where over the same period haptoglobin concentration increased (P < 0.05). Overall, our results show that the addition of AOX and HSCAS in the diet may alleviate the negative effects due to DON contamination on the antioxidant status and immune response of vaccinated weanling pigs.

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Variations in the major envelope glycoprotein GP5 of Czech strains of porcine reproductive and respiratory syndrome virus

Journal of General Virology ◽

10.1099/0022-1317-81-10-2497 ◽

2000 ◽

Vol 81 (10) ◽

pp. 2497-2502 ◽

Cited By ~ 38

Author(s):

Stanislav Indik ◽

Lubomír Valíček ◽

Dieter Klein ◽

Jana Klánová

Keyword(s):

Amino Acid ◽

Nucleotide Sequence ◽

Phylogenetic Tree ◽

Envelope Glycoprotein ◽

Point Mutations ◽

Nucleotide Sequences ◽

Respiratory Syndrome Virus

The major envelope glycoprotein genes (ORF5) of seven Czech isolates of porcine reproductive and respiratory syndrome virus (PRRSV) were amplified and their nucleotide sequences were determined. ORF5 displayed nucleotide and amino acid identities of 87·5–100% and 87·6–100%, respectively, among the isolates. In a phylogenetic tree, all European isolates were grouped in a genotype distinct from that of reference American strains (VR-2332, IAF-Klop). Among the European isolates, two different clades were identified. Two Czech isolates (V-501 and V-503) and Italian strain PRRSV 2156 fell into one clade. The remaining European strains comprised the second clade. Surprisingly, two separately clustered strains (V-501 and V-516) were isolated from the same herd. Additionally, the possible effect of in vitro cultivation on the nucleotide sequence was analysed. Nine point mutations in the ORF5 region resulted from 152 in vitro passages of the V-502 isolate in MARC-145 cells.

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Closing and Opening Holes in the Glycan Shield of HIV-1 Envelope Glycoprotein SOSIP Trimers Can Redirect the Neutralizing Antibody Response to the Newly Unmasked Epitopes

Journal of Virology ◽

10.1128/jvi.01656-18 ◽

2018 ◽

Vol 93 (4) ◽

Cited By ~ 12

Author(s):

Rajesh P. Ringe ◽

Pavel Pugach ◽

Christopher A. Cottrell ◽

Celia C. LaBranche ◽

Gemma E. Seabright ◽

...

Keyword(s):

Antibody Response ◽

Viral Entry ◽

Envelope Glycoprotein ◽

Neutralizing Antibodies ◽

Neutralizing Antibody ◽

Tier 2 ◽

Protein Surfaces ◽

Opening And Closing ◽

Glycan Site ◽

Hiv 1

ABSTRACTIn HIV-1 vaccine research, native-like, soluble envelope glycoprotein SOSIP trimers are widely used for immunizing animals. The epitopes of autologous neutralizing antibodies (NAbs) induced by the BG505 and B41 SOSIP trimers in rabbits and macaques have been mapped to a few holes in the glycan shields that cover most of the protein surfaces. For BG505 trimers, the dominant autologous NAb epitope in rabbits involves residues that line a cavity caused by the absence of a glycan at residue 241. Here, we blocked this epitope in BG505 SOSIPv4.1 trimer immunogens by knocking in an N-linked glycan at residue 241. We then opened holes elsewhere on the trimer by knocking out single N-linked glycans at residues 197, 234, 276, 332, and 355 and found that NAb responses induced by the 241-glycan-bearing BG505 trimers were frequently redirected to the newly opened sites. The strongest evidence for redirection of the NAb response to neoepitopes, through the opening and closing of glycan holes, was obtained from trimer immunogen groups with the highest occupancy of the N241 site. We also attempted to knock in the N289-glycan to block the sole autologous NAb epitope on the B41 SOSIP.v4.1 trimer. Although a retrospective analysis showed that the new N289-glycan site was substantially underoccupied, we found some evidence for redirection of the NAb response to a neoepitope when this site was knocked in and the N356-glycan site knocked out. In neither study, however, was redirection associated with increased neutralization of heterologous tier 2 viruses.IMPORTANCEEngineered SOSIP trimers mimic envelope-glycoprotein spikes, which stud the surface of HIV-1 particles and mediate viral entry into cells. When used for immunizing test animals, they elicit antibodies that neutralize resistant sequence-matched HIV-1 isolates. These neutralizing antibodies recognize epitopes in holes in the glycan shield that covers the trimer. Here, we added glycans to block the most immunogenic neutralization epitopes on BG505 and B41 SOSIP trimers. In addition, we removed selected other glycans to open new holes that might expose new immunogenic epitopes. We immunized rabbits with the various glycan-modified trimers and then dissected the specificities of the antibody responses. Thus, in principle, the antibody response might be diverted from one site to a more cross-reactive one, which would help in the induction of broadly neutralizing antibodies by HIV-1 vaccines based on envelope glycoproteins.

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Porcine Reproductive and Respiratory Syndrome Virus Promotes SLA-DR-Mediated Antigen Presentation of Nonstructural Proteins To Evoke a Nonneutralizing Antibody Response In Vivo

Journal of Virology ◽

10.1128/jvi.01423-20 ◽

2020 ◽

Vol 94 (21) ◽

Author(s):

Chunyan Wu ◽

Bingjun Shi ◽

Di Yang ◽

Kun Zhang ◽

Jie Li ◽

...

Keyword(s):

Antibody Response ◽

Antigen Presentation ◽

Wild Strain ◽

Effective Vaccine ◽

Respiratory Syndrome Virus ◽

Nonstructural Proteins ◽

Hyperimmune Serum ◽

Rapid Induction

ABSTRACT The humoral immune response against porcine reproductive and respiratory syndrome virus (PRRSV) infection is characterized by a rapid induction of nonneutralizing antibodies (non-NAbs) against nonstructural proteins (NSPs). Here, we systematically investigated the potential mechanism for the induction of PRRSV NSP-specific non-NAbs. Our data suggested that PRRSV NSP-specific antibodies appeared within 10 days after PRRSV infection in vivo. In the in vitro model, functional upregulation of swine leukocyte antigen (SLA)-DR was observed in bone marrow-derived dendritic cells (BMDCs) and porcine alveolar macrophages (PAMs), whereas remarkable inhibition at the mRNA level was observed after infection by both PRRSV-1 and PRRSV-2 isolates. Notably, the inconsistency in SLA-DR expression between the mRNA and protein levels resulted from deubiquitination of SLA-DR via the ovarian tumor (OTU) domain of PRRSV NSP2, which inhibited ubiquitin-mediated degradation. Moreover, mass spectrometry-based immunopeptidome analysis identified immunopeptides originating from multiple PRRSV NSPs within SLA-DR of PRRSV-infected BMDCs. Meanwhile, these PRRSV NSP-derived immunopeptides could be specifically recognized by serum from PRRSV-infected piglets. Notably, certain NSP-derived immunopeptides characterized in vitro could be identified from PAMs or hilar lymph nodes from PRRSV-infected piglets. More importantly, an in vitro neutralizing assay indicated that serum antibodies against NSP immunopeptides were unable to neutralize PRRSV in vitro. Conversely, certain structural protein (SP)-derived immunopeptides were identified and could be recognize by pig hyperimmune serum against PRRSV, which further indicates that the NSP-derived antibody response is nonprotective in vivo. In conclusion, our data suggested that PRRSV infection interferes with major histocompatibility complex class II (MHC-II) molecule-mediated antigen presentation in antigen-presenting cells (APCs) via promoting SLA-DR expression to present immunopeptides from PRRSV NSPs, which contributes to the induction of non-NAbs in vivo. IMPORTANCE PRRSV has haunted the swine industry for over 30 years since its emergence. Besides the limited efficacy of PRRSV modified live vaccines (MLVs) against heterogeneous PRRSV isolates, rapid induction of nonneutralizing antibodies (non-NAbs) against PRRSV NSPs after MLV immunization or wild-strain infection is one of the reasons why development of an effective vaccine has been hampered. By using in vitro-generated BMDCs as models to understand the antigen presentation process of PRRSV, we obtained data indicating that PRRSV infection of BMDCs promotes functional SLA-DR upregulation to present PRRSV NSP-derived immunopeptides for evoking a non-NAb response in vivo. Our work not only uncovered a novel mechanism for interference in host antigen presentation by PRRSV but also revealed a novel insight for understanding the rapid production of nonneutralizing antibodies against PRRSV NSPs, which may have benefit for developing an effective vaccine against PRRSV in the future.

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N-Linked Glycosylation of GP5 of Porcine Reproductive and Respiratory Syndrome Virus Is Critically Important for Virus ReplicationIn Vivo

Journal of Virology ◽

10.1128/jvi.07067-11 ◽

2012 ◽

Vol 86 (18) ◽

pp. 9941-9951 ◽

Cited By ~ 40

Author(s):

Zuzhang Wei ◽

Tao Lin ◽

Lichang Sun ◽

Yanhua Li ◽

Xiaoming Wang ◽

...

Keyword(s):

Antibody Response ◽

Cultured Cells ◽

Neutralizing Antibody ◽

Respiratory Syndrome Virus ◽

Virus Infectivity ◽

Serum Samples ◽

Virus Life Cycle ◽

Glycosylation Sites ◽

High Level

It has been proposed that the N-linked glycan addition at certain sites in GP5 of porcine reproductive and respiratory syndrome virus (PRRSV) is important for production of infectious viruses and viral infectivity. However, such specific N-linked glycosylation sites do not exist in some field PRRSV isolates. This implies that the existence of GP5-associated glycanper seis not vital to the virus life cycle. In this study, we found that mutation of individual glycosylation sites at N30, N35, N44, and N51 in GP5 did not affect virus infectivity in cultured cells. However, the mutants carrying multiple mutations at N-linked glycosylation sites in GP5 had significantly reduced virus yields compared with the wild-type (wt) virus. As a result, no viremia and antibody response were detected in piglets that were injected with a mutant without all N-linked glycans in GP5. These results suggest that the N-linked glycosylation of GP5 is critically important for virus replicationin vivo. The study also showed that removal of N44-linked glycan from GP5 increased the sensitivity of mutant virus to convalescent-phase serum samples but did not elicit a high-level neutralizing antibody response to wt PRRSV. The results obtained from the present study have made significant contributions to better understanding the importance of glycosylation of GP5 in the biology of PRRSV.

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