Induction of Interferon-α and Tumor Necrosis Factor-Related Apoptosis-Inducing Ligand in Human Blood Mononuclear Cells by Hemagglutinin-Neuraminidase but Not F Protein of Newcastle Disease Virus

Abstract

To determine molecular viral components which can induce innate immune responses in human peripheral blood mononuclear cells (PBMC), we investigated the anti-neoplastic agent Newcastle disease virus (NDV) and its two spike proteins hemagglutinin-neuraminidase (HN) and fusion protein (F). NDV was an excellent inducer in PBMC of IFN-alpha production and capable of inducing upregulation of plasma membrane expression of tumor necrosis factor related apoptosis inducing ligand (TRAIL). Viral replication was not required for these responses because NDV inactivated for 5 min by UV was as good as live NDV. NDV-modified and paraformaldehyde-fixed BHK cells could also trigger IFN-alpha and TRAIL induction, indicating that contacts of responder cells with NDV-modified cell surfaces are sufficient to induce these activities in PBMC. Antibodies against HN but not F were able to block these responses. Finally we could show that HN but not F induced IFN-alpha and TRAIL in PBMC. This was possible through the use of respective gene transfectants generated with the help of Semliki Forest virus (SFV) replicase-based DNA recombinant expression systems. Upon contact with BHK cells expressing HN but not F at their cell surface, human PBMC produced IFN-alpha and some cells, including monocytes and T lymphocytes, upregulated cell surface TRAIL expression.

A tumor vaccine containing anti‐CD3 and anti‐CD28 bispecific antibodies triggers strong and durable antitumor activity in human lymphocytes

“…3,4 Memory DTH responses could be reactivated with ATV-NDV vaccine many times, 5 thus indicating that no depletion of antitumor reactive T cells had occurred. We have postulated that the addition of danger signals through doublestranded RNA (dsRNA) 32 and cell surface-expressed HN proteins 10 can overcome T-cell unresponsiveness. It also possibly prevents activation-induced cell death (AICD) or unresponsiveness.…”

“…4 Infection of tumor cells with Newcastle Disease Virus (NDV) was shown to introduce into the tumor cells danger signals 6 by way of double-stranded RNA 7,8 and T-cell costimulatory activity 9 via the viral cell surface-expressed hemagglutinin-neuraminidase (HN) protein. 10 NDV can infect tumor cells efficiently, leading to high levels of HN expression on proliferating and nonproliferating cells because it replicates in the cytoplasm and does not integrate into the DNA. Further advantages of NDV are its broad applicability and its tumor-selective replication properties.…”

Recombinant Newcastle disease virus (NDV) with inserted gene coding for GM-CSF as a new vector for cancer immunogene therapy

“…36 Also a combination of IFN-a and GM-CSF secreting tumor vaccine was shown to significantly enhance the potency of the immunotherapeutic effect. 37 The expression of the HN protein of NDV at the cell surface of the ATV-NDV tumor vaccine 5 as well as the replication of NDV within these cells may be of central importance for the induction of IFN-a at the vaccination site and consequently for the observed antitumor response in clinical studies based on NDV tumor vaccine. 16 Here, we show that a tumor vaccine obtained by irradiation and infection with rec(GM-CSF) induces a stronger IFN-a response in human monocytes and PDCs than a vaccine infected with the virus without incorporated foreign gene.…”

“…2 Three cellular mechanisms have been proposed for its anti-neoplastic activities: (i) oncolysis: 3 oncolytic strains may simply kill tumor cells directly. (ii) Provision of danger signals: 4,5 replication of NDV occurs in the tumor cells' cytoplasm. It is associated with the production of single-and doublestranded viral RNA.…”

Adenovirus-mediated gene transfer of pathogen-associated molecular patterns for cancer immunotherapy

“…25 Immunostimulatory properties of the hemagglutininneuraminidase (HN) of the Newcastle disease virus (NDV) have been demonstrated in vitro. 26,27 Mouse fibroblasts transfected with HN or infected with NDV have been shown to stimulate specific T-cell proliferation with greater efficacy than untreated fibroblasts. 26 A major effect of NDV infection is the induction of IFN-a/b secretion, which can be mediated either through TLRs or by RIG-I-dependent signaling.…”