Factors affecting amplification of BK virus episomal vectors in human cells

M. P. Grossi, A. Caputo, L. Paolini, P. G. Balboni, G. Gerna, M. Pagnani, A. Corallini, G. Barbanti-Brodano

Research output: Contribution to journalArticle

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Abstract

Analysis of factors determining replication of BK virus (BKV) episomal vectors in human cells showed that vector copy number was related to the level of BKV T antigen expression. T antigen was synthesized efficiently, as assessed by indirect immunofluorescence, in vector-transfected primary embryonic fibroblasts undergoing neoplastic transformation. Surprisingly, transfected continuous cell lines (143B, HeLa and KB), kept under biochemical selection or tested in transient assays, produced negligible amounts or no T antigen, revealed only by a sensitive ELISA test, suggesting that in these cells vector amplification was under the control of cellular factors. Presence or absence of BKV late region sequences, BKV strain, orientation of the inserted genes and presence or absence of selection were not relevant for vector replication. Type of biochemical selection, however, was important, since BKV vectors containing the thymidine kinase gene replicated better than those containing the neo gene. Despite great variability, vector copy number increased in transfected clones of adenovirus 5-transformed 293 cells, in the absence of immunofluorescence detectable T antigen. These cells express adenovirus immediate early proteins E 1 A and E 1 B which may directly or indirectly activate BKV origin of replication.

Original languageEnglish
Pages (from-to)249-259
Number of pages11
JournalArchives of Virology
Volume99
Issue number3-4
DOIs
Publication statusPublished - Sep 1988

Fingerprint

BK Virus
Viral Tumor Antigens
Adenovirus Early Proteins
Immediate-Early Proteins
Genes
Replication Origin
Thymidine Kinase
Indirect Fluorescent Antibody Technique
Virus Replication
Adenoviridae
Statistical Factor Analysis
Fluorescent Antibody Technique
Clone Cells
Fibroblasts
Enzyme-Linked Immunosorbent Assay
Cell Line

ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology
  • Genetics

Cite this

Grossi, M. P., Caputo, A., Paolini, L., Balboni, P. G., Gerna, G., Pagnani, M., ... Barbanti-Brodano, G. (1988). Factors affecting amplification of BK virus episomal vectors in human cells. Archives of Virology, 99(3-4), 249-259. https://doi.org/10.1007/BF01311074

Factors affecting amplification of BK virus episomal vectors in human cells. / Grossi, M. P.; Caputo, A.; Paolini, L.; Balboni, P. G.; Gerna, G.; Pagnani, M.; Corallini, A.; Barbanti-Brodano, G.

In: Archives of Virology, Vol. 99, No. 3-4, 09.1988, p. 249-259.

Research output: Contribution to journalArticle

Grossi, MP, Caputo, A, Paolini, L, Balboni, PG, Gerna, G, Pagnani, M, Corallini, A & Barbanti-Brodano, G 1988, 'Factors affecting amplification of BK virus episomal vectors in human cells', Archives of Virology, vol. 99, no. 3-4, pp. 249-259. https://doi.org/10.1007/BF01311074
Grossi, M. P. ; Caputo, A. ; Paolini, L. ; Balboni, P. G. ; Gerna, G. ; Pagnani, M. ; Corallini, A. ; Barbanti-Brodano, G. / Factors affecting amplification of BK virus episomal vectors in human cells. In: Archives of Virology. 1988 ; Vol. 99, No. 3-4. pp. 249-259.
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