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Evaluation of IRES-mediated, cell-type-specific cytotoxicity of poliovirus using a colorimetric cell proliferation assay

  1. Author:
    Yang, X. Y.
    Chen, E. Y.
    Jiang, H. G.
    Muszynski, K.
    Harris, R. D.
    Giardina, S. L.
    Gromeier, M.
    Mitra, G.
    Soman, G.

  2. Author Address

    Yang, Xiaoyi, Chen, Eying, Jiang, Hengguang, Harris, Raymond D.; Giardina, Steven L.; Mitra, Gautam, Soman, Gopalan] SAIC Frederick Inc, Bioanalyt Dev Lab, Biopharmaceut Dev Program, NCI Frederick, Frederick, MD 21702 USA. [Muszynski, Karen] Natl Canc Inst, Biol Resource Branch, Frederick, MD 21702 USA. [Gromeier, Matthias] Duke Univ, Med Ctr, Div Neurosurg, Dept Surg, Durham, NC 27710 USA.
    1. Year: 2009
    2. Date: Jan
  2. Journal: Journal of Virological Methods
    1. 155
    2. 1
    3. Pages: 44-54
  4. Type of Article: Article
  1. Abstract:

    PVS-RIPO is it recombinant oncolytic poliovirus designed for clinical application to target CD155 expressing malignant gliomas and other malignant diseases. PVS-RIPO does not replicate in healthy neurons and is therefore non-pathogenic in rodent and non-human primate models of poliomyelitis. A tetrazolium salt dye-based cellular assay was developed and qualified to define the cytotoxicity of virus preparations on susceptible cells and to explore the target cell specificity of PVS-RIPO. In this assay, PVS-RIPO inhibited proliferation of U87-MG astrocytoma cells in a dose-dependent manner. However, HEK293 cells were much less susceptible to cell killing by PVS-RIPO. In contrast, the Sabin type 1 live attenuated poliovirus vaccine strain (PV(1)S) was cytotoxic to both HEK293 and U87-MG cells. The correlation between expression of CD155 and cytotoxicity was also explored using six different cell lines. There was little or no expression of CD155 and PVS-RIPO-induced cytotoxicity in Jurkat and Daudi cells. HEK293 was the only cell line tested that showed CD155 expression and resistance to PVS-RIPO cytotoxicity. The results indicate that differential cytotoxicity measured by the colorimetric assay can be used to evaluate the cytotoxicity and cell-type specificity of recombinant strains of poliovirus and to demonstrate lot to lot consistency during the manufacture of viruses intended for clinical use. (c) 2008 Elsevier B.V. All rights reserved.

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  2. DOI: 10.1016/j.jviromet.2008.09.020
  3. PMID: 18951922
  4. PMCID: PMC2650828

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