A type-specific serological test to distinguish antibodies to equine herpesviruses 4 and 1

Crabb, Brendan S.; MacPherson, C. M.; Reubel, G. H.; Browning, Glenn F.; Studdert, M. J.; Drummer, Heidi E.

doi:10.1007/bf01309860

Cited by 93 publications

(65 citation statements)

References 18 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Another important feature of gG is the induction of a strong Ab response in response to infection (10,34). Consequently, one has to consider the effect of Abs directed against gG for its ability to properly function as a vCKBP.…”

Section: Discussionmentioning

confidence: 99%

Herpesvirus Chemokine-Binding Glycoprotein G (gG) Efficiently Inhibits Neutrophil Chemotaxis In Vitro and In Vivo

Walle

May

Sukhumavasi

et al. 2007

The Journal of Immunology

View full text Add to dashboard Cite

Glycoprotein G (gG) of alphaherpesviruses has been described to function as a viral chemokine-binding protein (vCKBP). More recently, mutant viruses devoid of gG have been shown to result in increased virulence, but it remained unclear whether the potential of gG to serve as a vCKBP is responsible for this observation. In this study, we used equine herpesvirus type 1 (EHV-1) as a model to study the pathophysiological importance of vCKBP activity. First, in vitro chemotaxis assays studying migration of immune cells, an important function of chemokines, were established. In such assays, supernatants of EHV-1-infected cells significantly inhibited IL-8-induced chemotaxis of equine neutrophils. Identification of gG as the responsible vCKBP was achieved by repeating similar experiments with supernatants from cells infected with a gG-negative mutant, which were unable to alter IL-8-induced equine neutrophil migration. Furthermore, rEHV-1 gG was able to significantly reduce neutrophil migration, establishing gG as a bona fide vCKBP. Second, and importantly, in vivo analyses in a murine model of EHV-1 infection showed that neutrophil migration in the target organ lung was significantly reduced in the presence of gG. In summary, we demonstrate for the first time that EHV-1 gG not only binds to chemokines but is also capable of inhibiting their chemotactic function both in vitro and in vivo, thereby contributing to viral pathogenesis and virulence.

show abstract

Section: Discussionmentioning

confidence: 99%

Herpesvirus Chemokine-Binding Glycoprotein G (gG) Efficiently Inhibits Neutrophil Chemotaxis In Vitro and In Vivo

Walle

May

Sukhumavasi

et al. 2007

The Journal of Immunology

View full text Add to dashboard Cite

show abstract

“…9,14 The pGEX-4T expression system has been used by several investigators to develop ELISAs for the detection of antibodies targeted against viral proteins carrying major antigenic determinants. 5,7,9,30 In the present study, several porcine sera were screened for the presence of antibodies against ORFs 3, 4, 5, and 7 products of PRRSV. The purposes of the study were 1) to determine the antigenicity of those proteins (especially for ORFs 3 and 4) and 2) to confirm that antibodies to the GP 5 are indeed responsible for VN in natural PRRSV infection.…”

Section: Discussionmentioning

confidence: 99%

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

et al. 1999

View full text Add to dashboard Cite

Abstract. To determine the structural protein of the porcine reproductive and respiratory syndrome virus (PRRSV) involved in the production of neutralizing antibodies following clinical infection, correlation was studied between virus neutralization capability of convalescent pig sera and antibody response to the open reading frames (ORFs) 3-, 4-, 5-, and 7-encoded proteins GP 3 , GP 4 , GP 5 , and N, respectively. Individual virus genes were cloned into the pGEX-4T-1 vector, and the recombinant viral proteins were expressed in Escherichia coli fused to the glutathione S-transferase (GST) protein. The resulting GST-ORF3, GST-ORF4, GST-ORF5, and GST-ORF7 recombinant fusion proteins were purified by electroelution and used as antigens for serologic testing by indirect enzyme-linked immunosorbent assay and western immunoblotting. The overall antibody (IgG and IgM) titers to PRRSV of pooled convalescent pig sera were first determined by indirect immunofluorescence, and then sera with specific IgG titers Ͼ1:1,024 were tested for their specific virus neutralization activity and reactivity to individual recombinant fusion proteins. Except for the early immune response (as revealed by the presence of specific IgM), neutralizing titers were correlated with anti-GP 5 titers but not with anti-GP 3 and anti-GP 4 titers. The correlation between virus neutralization and anti-GP 5 titers was significant (r ϭ 0.811, P Յ 0.001).Porcine reproductive and respiratory syndrome virus (PRRSV) is an economically important pig pathogen structurally and genomically related to the equine arteritis virus (EAV), the murine lactate dehydrogenase-elevating virus, and the simian hemorrhagic fever virus. 20,28 These viruses have been recently classified in the genus Arterivirus, family Arteriviridae in the order Nidovirales, which also comprises members of the family Coronaviridae. 3 The genome of the PRRSV is a positive single-stranded polyadenylated RNA molecule approximately 15 kb in length containing 8 open reading frames (ORFs), which are transcribed in the infected cells as a nested set of subgenomic mRNAs. 8,20 The enveloped viral particles contain 3 major structural proteins, a glycosylated envelope protein of 25 kD (GP 5 ), an unglycosylated membrane (M) protein of 19 kD, and a nucleocapsid (N) protein of 14 kD, encoded by ORFs 5, 6, and 7, respectively. 16,17,21 The ORFs 2, 3, and 4 of the Lelystad virus, the European prototype of PRRSV, encode for putative membrane-associated glycoproteins, GP 2 , GP 3 , and GP 4 , respectively, with respective molecular masses of 30, 45, and 31 kD. 21 Kinetics of development of antiviral and neutralizing antibodies to PRRSV following clinical infection in pigs have been previously studied. 15,24 The deter- mination of the protein(s) responsible for the induction of neutralizing antibodies is a key element to consider for the development of an efficient recombinant vaccine. Several reports clearly established that the ectodomain of the G L glycoprotein of EAV, the counterpart of the GP 5 of PRRSV, co...

show abstract

“…ELISA was carried out as described previously (Crabb et al, 1995), with the exception that wells were coated with 1 mg ml 21 purified ERAV.393/76 and that bound rabbit antibodies were detected using a 1 : 1000 dilution of . Purified fusion proteins were separated by 12 % SDS-PAGE under reducing conditions and stained with Coomassie blue or transferred to PVDF membranes (Immobilon, Millipore) and probed with ERAV.393/76 rabbit antisera diluted 1 : 1000 and horse ERAV antisera diluted 1 : 250 (horses C and G).…”

mentioning

confidence: 99%

Mapping epitopes in equine rhinitis A virus VP1 recognized by antibodies elicited in response to infection of the natural host

Stevenson

Hartley

Huang

et al. 2003

Journal of General Virology

View full text Add to dashboard Cite

Equine rhinitis A virus (ERAV) is an important respiratory pathogen of horses and is of additional interest because of its close relationship and common classification with foot-and-mouth disease virus (FMDV). As is the case with FMDV, the VP1 capsid protein of ERAV has been shown to be a target of neutralizing antibodies. In FMDV VP1, such antibodies commonly recognize linear epitopes present in the bG-bH loop region. To map linear B cell epitopes in ERAV VP1, overlapping fragments spanning its length were expressed in Escherichia coli as glutathione S-transferase (GST) fusion proteins. These fusion proteins were tested for reactivity with sera from ERAV-infected horses and with polyclonal sera from ERAV-immunized rabbits and mice. Regions at the N-and C-termini as well as the bE-bF and the bG-bH loop regions contained B cell epitopes that elicited antibodies in the natural host. GST fusion proteins of these regions also elicited antibodies following immunization of rabbits and mice, which, in general, strongly recognized native ERAV VP1 but which were non-neutralizing. It is concluded that the N-terminal region of ERAV VP1, in particular, contains strong B cell epitopes.Equine rhinitis A virus (ERAV) is an important respiratory pathogen of horses. Disease is characterized by fever (41±0?5˚C) and clinical signs that include nasal discharge, coughing, anorexia, pharyngitis and lymphadenitis (Plummer, 1962). ERAV is classified with foot-and-mouth disease virus (FMDV), albeit as a separate cluster, in the genus Aphthovirus, family Picornaviridae. ERAV shares many physico-chemical and biological properties with FMDV and the genome structures and sequences of the two viruses are also similar (Li et al., 1996;Newman et al., 1973;Plummer, 1963;Studdert & Gleeson, 1978;Wutz et al., 1996).Picornavirus capsid proteins VP1, VP2 and VP3 share structural homology and are composed of wedge-shaped, eight-stranded, b-barrels, which differ in the size and conformation of the connecting loops between their strands and the extensions of their N-and C-termini (Rueckert, 2001). The amino acid sequences of the loops that connect the b-strands and the N-and C-terminal regions that extend from the b-barrel domain give each picornavirus its distinct morphology and antigenicity (Mateu, 1995;Rueckert, 2001). For example, the surface loops of VP1, VP2 and VP3 of poliovirus type 1 (PV-1) and human rhinovirus type 14 capsid structures protrude from the virion surface to form the receptor-binding canyon, as well as the major antigenic sites of these viruses. In comparison, the VP1 protein within the relatively smooth surface of FMDV particles contains a linear peptide, the bG-bH loop, which is both the dominant neutralization epitope and is also involved in receptor binding to various integrins via an RGD motif (Berinstein et al., 1995;Danen et al., 1995;Jackson et al., 2000;Mateu et al., 1995;Stanway, 1990;Strohmaier et al., 1982).In marked contrast to FMDV and human rhinoviruses, the predicted amino acid sequence of ERAV P1 and, in particu...

show abstract

A type-specific serological test to distinguish antibodies to equine herpesviruses 4 and 1

Cited by 93 publications

References 18 publications

Herpesvirus Chemokine-Binding Glycoprotein G (gG) Efficiently Inhibits Neutrophil Chemotaxis In Vitro and In Vivo

Herpesvirus Chemokine-Binding Glycoprotein G (gG) Efficiently Inhibits Neutrophil Chemotaxis In Vitro and In Vivo

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

Mapping epitopes in equine rhinitis A virus VP1 recognized by antibodies elicited in response to infection of the natural host

Contact Info

Product

Resources

About

A type-specific serological test to distinguish antibodies to equine herpesviruses 4 and 1

Cited by 93 publications

References 18 publications

Herpesvirus Chemokine-Binding Glycoprotein G (gG) Efficiently Inhibits Neutrophil Chemotaxis In Vitro and In Vivo

Herpesvirus Chemokine-Binding Glycoprotein G (gG) Efficiently Inhibits Neutrophil Chemotaxis In Vitro and In Vivo

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

Mapping epitopes in equine rhinitis A virus VP1 recognized by antibodies elicited in response to infection of the natural host

Contact Info

Product

Resources

About

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