Induction of Nitric Oxide and TNF-Α in Newcastle Disease Virus (NDV) AF2240 Infected RAW 264.7 Macrophages and their Cytotoxic Activity on MDA-MB-231 Breast Cancer Cell Line

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.

Regression of solid breast tumours in mice by Newcastle disease virus is associated with production of apoptosis related-cytokines

“…Due to its unique mechanism of action and its ability to impact on many types of human cancer, NDV has been proposed as a potent anti-cancer agent. Many strains of NDV were found to exhibit anti-cancer activity, for instance, MTH-68/H and AF2240 exhibited its anti-tumour effect via induction of nitric oxide synthesis from macrophages [40, 41]. In addition, systemic therapy with NDV strain PV701 showed a promising result as an important therapy for patients with a solid tumour [42].…”

Antitumor and immunostimulatory activities of a genotype V recombinant attenuated veterinary Newcastle disease virus vaccine

“…Previous studies in vitro using the lytic NDV strain 73-T showed that human PBMC upregulate the expression of TNF-α in supernatant after 24 h of stimulation (18,42). Other reports have demonstrated the capability of NDV to stimulate the production of this cytokine, although it was in murine macrophages (19,43). In patients with advanced solid tumors, the administration of a single or multiple doses of the lentogenic NDV PV701 induced the rejection of the tumors with an increased amount in serum of pro-inflammatory cytokines, such as TNF-α (44).…”

Study of Newcastle Disease Virus as an Immunostimulatory Agent for Anticancer Effect

“…12,13 Other studies by Zeng et al and Ahmad et alhave carried out inactivation at UV lamp (245 nm, 2 mW/cm 2 for 30 mins and 15 mins respectively. 14,15 However, our methodology of UV inactivation at 254 nm, power 220µW/cm 2 at distance of 10 cm for 30 mins served the purpose as well as hemagglutinin activity was intact.…”