Expression of Aleutian mink disease parvovirus capsid proteins by a recombinant vaccinia virus: Self-assembly of capsid proteins into particles

D. L. Clemens, J. B. Wolfinbarger, S. Mori, B. D. Berry, S. F. Hayes, M. E. Bloom

Research output: Contribution to journalArticle

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Abstract

A portion of a cDNA clone containing coding sequences for both structural proteins (VP1 and VP2) of Aleutian mink disease parvovirus (ADV) was inserted into recombinant vaccinia viruses, VV:ADSP. Immunohistochemical staining of VV:ADSP-infected cells revealed that the ADV antigen was readily detected and localized in the nuclei of infected cells. Analysis of VV:ADSP-infected cell lysates indicated that both VP1 and VP2 were produced and comigrated with authentic VP1 and VP2 from ADV-infected Crandell feline kidney cells. These results suggested, therefore, that both VP1 and VP2 were synthesized from a single cloned transcript. CsCl density gradient centrifugation of partially purified VV:ADSP-infected cell lysates indicated that the majority of the antigen was located in a fraction with a density near 1.33 g/ml, indicative of empty ADV particles. Subsequent electron microscopic examination revealed the presence of 27-nm icosahedral virion-like structures at the same density, suggesting that the proteins self-assembled into empty virions. Furthermore, sera from eight of eight mice inoculated with VV:ADSP contained ADV-specific antibodies and two of these eight serum samples had neutralizing activity, indicating that the particles produced in VV:ADSP-infected cells were immunogenic. Finally, when lysates from VV:ADSP-infected cells were compared with standard ADV antigens in counterimmunoelectrophoresis assays, a similar pattern of specific reactivity was observed for sera from normal and infected mink.

Original languageEnglish (US)
Pages (from-to)3077-3085
Number of pages9
JournalJournal of virology
Volume66
Issue number5
StatePublished - Jan 1 1992

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Aleutian Mink Disease Virus
Aleutian mink disease virus
Virus Assembly
Vaccinia virus
Capsid Proteins
coat proteins
antigens
virion
cells
Antigens
Virion
counterimmunoelectrophoresis
Serum
Counterimmunoelectrophoresis
mink
structural proteins
Mink
cell nucleus
kidney cells
Density Gradient Centrifugation

ASJC Scopus subject areas

  • Microbiology
  • Immunology
  • Insect Science
  • Virology

Cite this

Expression of Aleutian mink disease parvovirus capsid proteins by a recombinant vaccinia virus : Self-assembly of capsid proteins into particles. / Clemens, D. L.; Wolfinbarger, J. B.; Mori, S.; Berry, B. D.; Hayes, S. F.; Bloom, M. E.

In: Journal of virology, Vol. 66, No. 5, 01.01.1992, p. 3077-3085.

Research output: Contribution to journalArticle

Clemens, D. L. ; Wolfinbarger, J. B. ; Mori, S. ; Berry, B. D. ; Hayes, S. F. ; Bloom, M. E. / Expression of Aleutian mink disease parvovirus capsid proteins by a recombinant vaccinia virus : Self-assembly of capsid proteins into particles. In: Journal of virology. 1992 ; Vol. 66, No. 5. pp. 3077-3085.
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