Abstract
Background: It is well known that Newcastle Disease Virus (NDV) AF2240 has antitumor activity against breast cancer cell lines. Several studies have thought this activity depend on direct viral oncolysis, which may not be the only factor that play a role in the antitumor effect of the virus. However, the mechanism underlying its antitumor effect is largely unresolved. In line with this observation, we postulated that NDV AF2240 may induce an in vitro production of Nitric Oxide (NO) and Tumor Necrosis Factor (TNF-α) in RAW 264.7 macrophages for cytotoxic activity on breast cancer cells. Methods: NDV AF2240 were subjected to different experimental conditions (Live, UV-inactivated and Heatinactivated), different Heamagglutination (HA) titer of the virus were co-cultured with RAW 264.7 cells to get the cell-free supernatant. Levels of TNF-α, and NO production were measured by ELISA and Griess assay respectively. MTT assay was later used to determine their cytotoxicity on MDA-MB-231 breast cancer cell line in vitro. Results: NDV was found to be an excellent inducer in RAW 264.7 macrophage cells of TNF-α (P<0.05) and NO production (P<0.05) when compared with the negative control. The supernatant containing NO and TNF-α was also found to be cytotoxic against MDA-MB-231 cells (P<0.05) when compared with the negative control, indicating that viral replication was not necessary for their production, because UV-inactivated NDV for 15 minutes was almost as good as lived NDV. However, heat inactivated NDV did not induce TNF-α and NO production. Conclusion: This suggests the relevance of TNF-α and NO in the indirect antitumor effect of NDV AF2240, and also shown that mare contact between the macrophages and the NDV is enough to induce TNF-α and NO production in macrophages.
Highlights
Newcastle disease virus (NDV) has recently been given more attention because of its antineoplastic and immunostimulatory properties [1,2]
In order to investigate whether Newcastle Disease Virus (NDV) AF2240 is capable of inducing TNF-α production, enzyme linked immunosorbent assay (ELISA) was performed on the supernatant obtained from RAW 264.7 macrophages cells, which have been cultured in the presence of live NDV, UVinactivated NDV, heat inactivated NDV and in medium alone to serve as a control
NDV induces Nitric Oxide (NO) production in RAW 264.7 macrophages in vitro, after incubating the RAW macrophages in the presence of live NDV, the level of NO in the supernatant obtained was found to be increasing with increased in HA titre (Figure 2)
Summary
Newcastle disease virus (NDV) has recently been given more attention because of its antineoplastic and immunostimulatory properties [1,2]. NDV is a contagious and deadly viral disease of birds that causes large economic loses to poultry industries worldwide [5]. It is non-segmented, single stranded, negative sense RNA virus belongs to Avulavirus in the Paramyxoviridae family [6]. Several studies have thought this activity depend on direct viral oncolysis, which may not be the only factor that play a role in the antitumor effect of the virus. The mechanism underlying its antitumor effect is largely unresolved In line with this observation, we postulated that NDV AF2240 may induce an in vitro production of Nitric Oxide (NO) and Tumor Necrosis Factor (TNF-α) in RAW 264.7 macrophages for cytotoxic activity on breast cancer cells
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