Structural and functional analysis of orthopoxvirus epitopes with neutralizing monoclonal antibodies

Czerny, Claus-Peter; Mahnel, H.

doi:10.1099/0022-1317-71-10-2341

Cited by 72 publications

(57 citation statements)

References 44 publications

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“…For EEV, the difference in infectious titer between the deletion mutant and the controls after infection at 0.01 PFU/cell was approximately 100-fold by 48 and 72 h p.i., and a large difference remained even at 96 h p.i. To determine the proportion of this extracellular virus that was EEV, rather than IMV that had been released from cells at these late times p.i., virus in the supernatant was incubated with mouse MAb 5B4/2F2 directed against the IMV surface protein encoded by the A27L gene and that neutralizes IMV infectivity (10). Under these conditions the remaining infectivity of v⌬F12L, representing EEV, was still approximately 100-fold lower than that of wild-type or revertant viruses.…”

Section: Resultsmentioning

confidence: 99%

Vaccinia Virus F12L Protein Is Required for Actin Tail Formation, Normal Plaque Size, and Virulence

Zhang

Wilcock

Smith

2000

J Virol

109

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Vaccinia virus gene F12L is shown to encode a 65-kDa protein that is synthesized early and late during infection and that is not modified by glycosylation. Computational sequence comparison revealed that related proteins are encoded by all sequenced chordopoxviruses. A virus deletion mutant lacking the F12L gene (v⌬F12L) and a revertant virus with the F12L gene reinserted into the deletion mutant (vF12L-rev) were constructed and analyzed. A version of the F12L gene with a C-terminal amino acid tag derived from the influenza virus hemagglutinin and that is recognized by a monoclonal antibody was also inserted into the F12L locus of v⌬F12L. Loss of the F12L protein reduced the formation of IMV 2-fold, but there was a dramatic (99.5%) reduction in actin tail formation, and the levels of cell-associated enveloped virus and extracellular enveloped virus were reduced 8-to 11-fold and 7-fold, respectively. Consistent with the lack of actin tail formation, v⌬F12L produced a very small plaque. The v⌬F12L virus was severely attenuated in vivo, such that a dose of v⌬F12L 10,000-fold greater than the dose of wild-type virus that induced severe disease was unable to induce disease in mice infected intranasally.

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Section: Resultsmentioning

confidence: 99%

Vaccinia Virus F12L Protein Is Required for Actin Tail Formation, Normal Plaque Size, and Virulence

Zhang

Wilcock

Smith

2000

J Virol

109

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“…and clarified by centrifugation. After appropriate dilution, any contaminating IMV infectivity was neutralized by addition of monoclonal antibody (MAb) 5B4\2F2 (final dilution of 1\2560) against the 14 kDa fusion protein (A27L gene product) of IMV (Czerny & Mahnel, 1990). In the conditions used in this study, MAb 5B4\2F2 neutralized 93 % of purified IMV.…”

Section: Methodsmentioning

confidence: 99%

Intracellular and extracellular vaccinia virions enter cells by different mechanisms.

Vanderplasschen

Hollinshead

Smith

1998

Journal of General Virology

149

179

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Vaccinia virus (VV) produces two antigenically distinct infectious virions, intracellular mature virus (IMV) and extracellular enveloped virus (EEV). Structurally, EEV consists of an IMV with an additional outer membrane containing proteins that are absent from IMV. EEV is important for virus dissemination both in vitro and in vivo. Studies of EEV entry have been hampered by having two infectious virions and by the rupture of the EEV outer membrane in the majority of EEV virions during their purification. To overcome these problems, we have developed a novel approach to study VV entry that is based on confocal microscopy and does not require EEV purification. This assay relies on immunofluorescent staining and detection of individual, intracellular,

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“…Reactivity exists with antigenic sites localized on the 14-kd fusion protein, the 32-kd adsorption protein, and the A-type inclusion body (ATI) protein of OPV encoded by the vaccinia virus open reading frames (ORF) A27L, D8L, and A25L. 7,10 In this assay, the monkey isolates proved to be identical to key group I, consisting of vaccinia virus and CPV strains. To determine the species within the genus OPV, isolates of this special group were further differentiated by D8L polymerase chain reaction (PCR) analysis and cycle sequencing of this ORF.…”

mentioning

confidence: 99%

Fatal Poxvirus Outbreak in a Colony of New World Monkeys

Mätz‐Rensing¹,

Ellerbrok²,

Ehlers³

et al. 2006

Vet Pathol

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Abstract. An epizootic infection was observed in a colony of 80 New World monkeys consisting of various species including a group of marmosets and Saguinus species. During the summer and autumn of 2002, 30 animals died of unknown diseases. Six animals were sent to the German Primate Center for investigation of the cause of death. A complete pathologic and histologic investigation was carried out. The animals exhibited erosive-ulcerative lesions of the oral mucous membranes. Advanced stages of the disease were characterised by hemorrhagic lesions on the skin distributed randomly over the body, but principally on the face, scrotal region, soles, and palms. Electron microscopy revealed virus particles with orthopox-like morphology within intracytoplasmic inclusions in epithelial cells. The DNA samples from various tissues were analyzed by use of a set of orthopox virus-specific, real-time polymerase chain reaction assays. Amplification products were sequenced to define the virus more precisely. Sequencing confirmed the presence of an orthopox virus. Sequence data indicated that all six animals were infected with the same virus. Propagation of the virus on Vero cells resulted in a rapidly progressive cytopathogenic effect. Preliminary phylogenetic analyses of two genes revealed closest homology to cowpox viruses. The origin of this poxvirus outbreak remains unexplained, and the strain and genus of the virus need to be determined in detail.

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Structural and functional analysis of orthopoxvirus epitopes with neutralizing monoclonal antibodies

Cited by 72 publications

References 44 publications

Vaccinia Virus F12L Protein Is Required for Actin Tail Formation, Normal Plaque Size, and Virulence

Vaccinia Virus F12L Protein Is Required for Actin Tail Formation, Normal Plaque Size, and Virulence

Intracellular and extracellular vaccinia virions enter cells by different mechanisms.

Fatal Poxvirus Outbreak in a Colony of New World Monkeys

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