Immunoaffinity purification and characterization of the envelope protein E1 of hog cholera virus

Wensvoort, G.; Boonstra, J.; Bodzinga, B. G.

doi:10.1099/0022-1317-71-3-531

Cited by 64 publications

(36 citation statements)

References 21 publications

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“…Thus, the nine cysteines in the C-terminal half of E1 are not essential for the conformation and accessibility of epitopes on the Nterminal half of El. It has been shown that E1 forms homodimers as well as heterodimers with E3 (gp31) and that cysteines are involved in this dimerization process (Wensvoort et al, 1990;Thiel et al, 1991). Here, we showed that the N-terminally located cysteines are probably involved in the antigenic structure of El.…”

Section: Discussionmentioning

confidence: 99%

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Epitope mapping of envelope glycoprotein E1 of hog cholera virus strain Brescia

Rijn

Gennip

Meijer

et al. 1993

Journal of General Virology

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Four antigenic domains (A, B, C and D) on envelope glycoprotein E1 (gp51-54) of hog cholera virus strain Brescia have been specified by using 13 monoclonal antibodies (MAbs) that recognize non-conserved and conserved epitopes. It was shown that the non-conserved epitopes map to the N-terminal half of E1 by analysis of chimeric E1 proteins of strains Brescia and C. Conserved epitopes, however, could not be mapped using this approach. Here we describe mapping of both conserved and non-conserved epitopes on E1 by the use of an extensive set of single and double deletion mutants of E1 of strain Brescia. Deletion mutants were transiently expressed in COS1 cells and analysed by immunostaining with the 13 MAbs directed against strain Brescia and four MAbs directed against strain C. All MAbs bound to the N-terminal half of El, i.e. amino acids 690 to 866 encoded by the sequence of strain Brescia. Domain B and one epitope in domain C are located between residues 690 and 773. Other epitopes in domain C are located on an extended region, i.e. between residues 690 and 800. Conserved epitopes of domain A are mapped between residues 766 and 866, whereas the only non-conserved epitope in this domain is located between residues 766 and 813. Domain D, represented by one MAb, is located in the same region as this nonconserved epitope of domain A, i.e. between residues 766 and 800. The results suggest the presence of two distinct antigenic units on El, one consisting of domains B and C and the other consisting of domain A.

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Section: Discussionmentioning

confidence: 99%

“…The major antigenic protein of HCV is envelope glycoprotein E1 (Wensvoort et aL, 1990). The immune response against E1 alone is sufficient for protection, because vaccination with a recombinant pseudorabies virus (PRV) which expresses the E1 gene of HCV strain Brescia protects swine against hog cholera (van Zijl et aL, 1991).…”

Section: Introductionmentioning

confidence: 99%

Epitope mapping of envelope glycoprotein E1 of hog cholera virus strain Brescia

Rijn

Gennip

Meijer

et al. 1993

Journal of General Virology

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“…The nomenclature of the pestivirus proteins used in this report will be proposed to the International Committee on Taxonomy of Viruses by the Flaviviridae Study Group. The nomenclature of the pestivirus envelope proteins used by the CSFV research group in Lelystad in previous publications (Hulst et al, 1993(Hulst et al, , 1994Moormann et al, 1990 ;van Zijl et al, 1991 ;Wensvoort, 1989 ;Wensvoort et al, 1986Wensvoort et al, a, b, 1990 …”

Section: Detection Of Ementioning

confidence: 99%

“…Although all three glycoproteins are associated with the virus envelope , up to now only antibodies against the two largest glycoproteins have been detected in animals infected with pestiviruses (Kwang et al, 1992 ;Terpstra & Wensvoort, 1988). The most immunodominant protein present in the virus envelope of pestiviruses is E2 (Donis & Dubovi, 1987 ;Wensvoort et al, 1990). When E2 is used as a subunit vaccine, it elicits high levels of neutralizing antibodies and protects pigs from classical swine fever (Hulst et al, 1993 ;van Zijl et al, 1991).…”

Section: Introductionmentioning

confidence: 99%

Inhibition of pestivirus infection in cell culture by envelope proteins E(rns) and E2 of classical swine fever virus: E(rns) and E2 interact with different receptors.

Hulst¹,

Moormann²

1997

Journal of General Virology

125

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Pure preparations of envelope glycoproteins E rns and E2 of classical swine fever virus (CSFV) synthesized in insect cells were used to study infection of porcine and bovine cells with the pestiviruses CSFV and bovine viral diarrhoea virus (BVDV). Almost 100 % inhibition of infection of porcine kidney cells with CSFV was produced by 100 µg/ml E rns . After removal of the virus no E rns was needed in the overlay medium (growth medium) to maintain this level of inhibition. In contrast, 100 % inhibition of infection of porcine kidney cells with CSFV by 10 µg/ml E2 was only achieved when E2 was added to the overlay medium. When E2 was omitted, a maximum of 50 % inhibition was achieved. This indicated that after the virus and E2 were removed from the cells, infection still occurred, by virus

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“…The surface structure of pestivirus virions is composed of three glycoproteins, E rns , E1, and E2 (46). E2 is present as a homodimer and as an E2-E1 heterodimer (46,51). The amino acid C terminus of E2 (and probably of E1 as well) functions as a membrane-spanning domain (14) and anchors these E2-E1 and E2-E2 dimers in the viral lipid membrane.…”

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confidence: 99%

Interaction of Classical Swine Fever Virus with Membrane-Associated Heparan Sulfate: Role for Virus Replication In Vivo and Virulence

Hulst¹,

Gennip²,

Vlot³

et al. 2001

J Virol

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Passage of native classical swine fever virus (CSFV) in cultured swine kidney cells (SK6 cells) selects virus variants that attach to the surface of cells by interaction with membrane-associated heparan sulfate (HS).Animal experiments indicated that this adaptive Ser-to-Arg mutation had no effect on the virulence of CSFV. Analysis of viruses reisolated from pigs infected with these recombinant viruses indicated that replication in vivo introduced no mutations in the genes of the envelope proteins E rns , E1, and E2. However, the blood of one of the three pigs infected with the S-RT virus contained also a low level of virus particles that, when grown under a methylcellulose overlay, produced relative large plaques, characteristic of an HS-independent virus. Sequence analysis of such a large-plaque phenotype showed that Arg 476 was mutated back to Ser 476 . Removal of HS from the cell surface and addition of heparin to the medium inhibited infection of cultured (SK6) and primary swine kidney cells with S-ST virus reisolated from pigs by about 70% whereas infection with the administered S-ST recombinant virus produced in SK6 cells was not affected. Furthermore, E rns S-ST protein, produced in insect cells, could bind to immobilized heparin and to HS chains on the surface of SK6 cells. These results indicated that S-ST virus generated in pigs is able to infect cells by an HS-dependent mechanism. Binding of concanavalin A (ConA) to virus particles stimulated the infection of SK6 cells with S-ST virus produced in these cells by12-fold; in contrast, ConA stimulated infection with S-ST virus generated in pigs no more than 3-fold. This suggests that the surface properties of S-ST virus reisolated from pigs are distinct from those of S-ST virus produced in cell culture. We postulate that due to these surface properties, in vivo-generated CSFV is able to infect cells by an HS-dependent mechanism. Infection studies with the HS-dependent S-RT virus, however, indicated that interaction with HS did not mediate infection of lung macrophages, indicating that alternative receptors are also involved in the attachment of CSFV to cells.

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Immunoaffinity purification and characterization of the envelope protein E1 of hog cholera virus

Cited by 64 publications

References 21 publications

Epitope mapping of envelope glycoprotein E1 of hog cholera virus strain Brescia

Epitope mapping of envelope glycoprotein E1 of hog cholera virus strain Brescia

Inhibition of pestivirus infection in cell culture by envelope proteins E(rns) and E2 of classical swine fever virus: E(rns) and E2 interact with different receptors.

Interaction of Classical Swine Fever Virus with Membrane-Associated Heparan Sulfate: Role for Virus Replication In Vivo and Virulence

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