| Author | Oliveira, Edson R. A. de | |
| Author | Lima, Bruna M. M. P. | |
| Author | Moura, Wlamir C. de | |
| Author | Nogueira, Ana Cristina M. de A. | |
| Access date | 2016-01-05T16:21:47Z | |
| Available date | 2016-01-05T16:21:47Z | |
| Document date | 2013 | |
| Citation | OLIVEIRA, Edson R. A. de; et al. Reduction of cell viability induced by IFN-alpha generates impaired data on antiviral assay using Hep-2C cells. Journal of Immunological Methods, v. 400–401, p.97–105, Dec. 2013. | pt_BR |
| ISSN | 0022-1759 | |
| URI | https://www.arca.fiocruz.br/handle/icict/12478 | |
| Language | eng | pt_BR |
| Publisher | Elsevier | pt_BR |
| Rights | restricted access | |
| Title | Reduction of cell viability induced by IFN-alpha generates impaired data on antiviral assay using Hep-2C cells | pt_BR |
| Type | Article | |
| DOI | 10.1016/j.jim.2013.10.011 | |
| Abstract | Type I interferons (IFNs) exert an array of important biological functions on the innate immune
response and has become a useful tool in the treatment of various diseases. An increasing
demand in the usage of recombinant IFNs, mainly due to the treatment of chronic hepatitis C
infection, augmented the need of quality control for this biopharmaceutical. A traditional
bioassay for IFN potency assessment is the cytopathic effect reduction antiviral assay where a
given cell line is preserved by IFN from a lytic virus activity using the cell viability as a frequent
measure of end point. However, type I IFNs induce other biological effects such as cell-cycle
arrest and apoptosis that can influence directly on viability of many cell lines. Here, we
standardized a cytopathic effect reduction antiviral assay using Hep-2C cell/mengovirus
combination and studied a possible impact of cell viability variations caused by IFN-alpha 2b on
responses generated on the antiviral assay. Using the four-parameter logistic model, we
observed less correlation and less linearity on antiviral assay when responses from IFN-alpha 2b
1000 IU/ml were considered in the analysis. Cell viability tests with MTT revealed a clear cell
growth inhibition of Hep-2C cells under stimulation with IFN-alpha 2b. Flow cytometric
cell-cycle analysis and apoptosis assessment showed an increase of S + G2 phase and higher
levels of apoptotic cells after treatment with IFN-alpha 2b 1000 IU/ml under our standardized
antiviral assay procedure. Considering our studied dose range, we also observed strong STAT1
activation on Hep-2C cells after stimulation with the higher doses of IFN-alpha 2b. Our findings
showed that the reduction of cell viability driven by IFN-alpha can cause a negative impact on
antiviral assays. We assume that the cell death induction and the cell growth inhibition effect of
IFNs should also be considered while employing antiviral assay protocols in a quality control
routine and emphasizes the importance of new approaches for IFN potency determination. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Biotecnologia e Fisiologia de Infecções Virais. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Nacional de Controle de Qualidade em Saúde. Programa de Pós-Graduação em Vigilância Sanitária. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Nacional de Controle de Qualidade em Saúde. Departamento de Imunologia. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Biotecnologia e Fisiologia de Infecções Virais. Rio de Janeiro, RJ, Brasil / Fundação Oswaldo Cruz. Instituto Nacional de Controle de Qualidade em Saúde. Programa de Pós-Graduação em Vigilância Sanitária. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Subject | IFN-alpha | pt_BR |
| Subject | Antiviral assay | pt_BR |
| Subject | Cell viability | pt_BR |
| Subject | Hep-2C | pt_BR |
| DeCS | Interferons | pt_BR |
| DeCS | Sobrevivência Celular | pt_BR |
