Artikel
Lack of toxicity of oncolytic Parvovirus H-1 in untransformed adult human brain cells
Parvovirus H-1: Onkoselektivität im humanen Hirngewebe
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Autoren
Veröffentlicht: | 4. Mai 2005 |
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Gliederung
Text
Objective
We already proved the efficient oncolytic effect of Parvovirus H-1 in human glioblastoma cell lines as well as in short-term cultures of glioblastomas and one gliosarcoma. For a variety of human cells but not for normal glial cells, Parvovirus H-1 is known to show limited-to-no cytocidal activity. In this study we report about the effects of H-1 virus infection on non-transformed human glial short-term cultures as well as on adult human organotypic brain cultures.
Methods
Specimens of adult human brain were obtained during epilepsy surgery. Isolation, culture and identification of glial cells was performed following the protocol of Brewer et al. 2001. Organotypic brain cultures were obtained following the method of Jung et al. 2001. Monolayers of glial cell cultures were inoculated with H-1 virus. Growth curves were recorded from infected and mock-treated cells. A qualitative analysis of the biosynthesis of all viral macromolecular components was performed. Organotypic cultures were inoculated with H-1 virus. Infected and mock-treated cultures were harvested on day 1,2,3,4,5 after infection and fixed in 4% PFA for histological examination.
Results
We were able to detect a synthesis of all viral macromolecular components in glial short-term cultures but there was no efficient cytolytic effect of H-1 virus infection. Growth curves of infected and mock-treated cultures showed no differences at a multiplicity of infection (MOI) of 1 plaque forming unit (pfu)/cell. At a MOI of 10 pfu/cell the infected cultures showed growth arrest in comparison to their mock-treated counterparts, but no altered morphology or cell lysis, maintaining an equal number of cells on day 1,3 and 6 after infection compared to the initial cell count. The histological examination of the organotypic cultures did not show signs of tissue damage on day 1,2,3,4 and 5 after infection with H-1 virus.
Conclusions
Infection of adult glial short-term cultures with parvovirus H-1 leads to viral replication but not to cytotoxic effects. Furthermore, infection of organotypic adult human brain cultures did not reveal any signs of tissue damage. This data supports the oncoselectivity of Parvovirus H-1, making it a promising candidate for oncolytic viral therapy of gliomas.