Long-Term and Memory Immune Responses in Mice against Newcastle Disease Virus-Like Particles Containing Respiratory Syncytial Virus Glycoprotein Ectodomains

ABSTRACT Respiratory syncytial virus (RSV) is the leading cause of serious respiratory infections in children as well as a serious cause of disease in elderly and immunosuppressed populations. There are no licensed vaccines available to prevent RSV disease. We have developed a virus-like particle (VLP) vaccine candidate for protection from RSV. The VLP is composed of the NP and M proteins of Newcastle disease virus (NDV) and a chimeric protein containing the cytoplasmic and transmembrane domains of the NDV HN protein and the ectodomain of the human RSV G protein (H/G). Immunization of mice with 10 or 40 μg total VLP-H/G protein by intraperitoneal or intramuscular inoculation stimulated antibody responses to G protein which were as good as or better than those stimulated by comparable amounts of UV-inactivated RSV. Immunization of mice with two doses or even a single dose of these particles resulted in the complete protection of mice from RSV replication in the lungs. Immunization with these particles induced neutralizing antibodies with modest titers. Upon RSV challenge of VLP-H/G-immunized mice, no enhanced pathology in the lungs was observed, although lungs of mice immunized in parallel with formalin-inactivated RSV (FI-RSV) showed the significant pathology that has previously been documented after immunization with FI-RSV. Thus, the VLP-H/G candidate vaccine was immunogenic in BALB/c mice and prevented replication of RSV in murine lungs, with no evidence of immunopathology. These data support further development of virus-like particle vaccine candidates for protection against RSV.

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Protection against Respiratory Syncytial Virus by a Recombinant Newcastle Disease Virus Vector

Journal of Virology ◽

10.1128/jvi.80.3.1130-1139.2006 ◽

2006 ◽

Vol 80 (3) ◽

pp. 1130-1139 ◽

Cited By ~ 88

Author(s):

Luis Martinez-Sobrido ◽

Negin Gitiban ◽

Ana Fernandez-Sesma ◽

Jerome Cros ◽

Sara E. Mertz ◽

...

Keyword(s):

Respiratory Syncytial Virus ◽

Dendritic Cell ◽

Newcastle Disease Virus ◽

Disease Virus ◽

Newcastle Disease ◽

Dendritic Cell Maturation ◽

Cell Maturation ◽

Viral Vaccine ◽

Fusion Glycoprotein ◽

Syncytial Virus

ABSTRACT Respiratory syncytial virus (RSV) is a major cause of severe lower respiratory tract disease in infants and the elderly, but no safe and effective RSV vaccine is yet available. For reasons that are not well understood, RSV is only weakly immunogenic, and reinfection occurs throughout life. This has complicated the search for an effective live attenuated viral vaccine, and past trials with inactivated virus preparations have led to enhanced immunopathology following natural infection. We have tested the hypothesis that weak stimulation of innate immunity by RSV correlates with ineffective adaptive responses by asking whether expression of the fusion glycoprotein of RSV by Newcastle disease virus (NDV) would stimulate a more robust immune response to RSV than primary RSV infection. NDV is a potent inducer of both alpha/beta interferon (IFN-α/β) production and dendritic cell maturation, while RSV is not. When a recombinant NDV expressing the RSV fusion glycoprotein was administered to BALB/c mice, they were protected from RSV challenge, and this protection correlated with a robust anti-F CD8+ T-cell response. The effectiveness of this vaccine construct reflects the differential abilities of NDV and RSV to promote dendritic cell maturation and is retained even in the absence of a functional IFN-α/β receptor.

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Assembly and Biological and Immunological Properties of Newcastle Disease Virus-Like Particles

Journal of Virology ◽

10.1128/jvi.01931-09 ◽

2010 ◽

Vol 84 (9) ◽

pp. 4513-4523 ◽

Cited By ~ 47

Author(s):

Lori W. McGinnes ◽

Homer Pantua ◽

Jason P. Laliberte ◽

Kathryn A. Gravel ◽

Surbhi Jain ◽

...

Keyword(s):

Immune Responses ◽

Newcastle Disease Virus ◽

Disease Virus ◽

Newcastle Disease ◽

Intracellular Cytokine Staining ◽

Nipah Virus ◽

Vaccine Virus ◽

Enzyme Linked Immunosorbent Assay ◽

Virus Like Particles ◽

Red Blood Cell Membranes

ABSTRACT Virus-like particles (VLPs) released from avian cells expressing the Newcastle disease virus (NDV) strain AV proteins NP, M, HN (hemagglutinin-neuraminidase), and F were characterized. The VLP-associated HN and F glycoproteins directed the attachment of VLPs to cell surfaces and fusion of VLP membranes with red blood cell membranes, indicating that they were assembled into VLPs in an authentic conformation. These particles were quantitatively prepared and used as an immunogen, without adjuvant, in BALB/c mice. The resulting immune responses, detected by enzyme-linked immunosorbent assay (ELISA), virus neutralization, and intracellular cytokine staining, were comparable to the responses to equivalent amounts of inactivated NDV vaccine virus. HN and F proteins from another strain of NDV, strain B1, could be incorporated into these VLPs. Foreign peptides were incorporated into these VLPs when fused to the NP or HN protein. The ectodomain of a foreign glycoprotein, the Nipah virus G protein, fused to the NDV HN protein cytoplasmic and transmembrane domains was incorporated into ND VLPs. Thus, ND VLPs are a potential NDV vaccine candidate. They may also serve as a platform to construct vaccines for other pathogens.

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Optimization of Human Immunodeficiency Virus Gag Expression by Newcastle Disease Virus Vectors for the Induction of Potent Immune Responses

Journal of Virology ◽

10.1128/jvi.01443-08 ◽

2008 ◽

Vol 83 (2) ◽

pp. 584-597 ◽

Cited By ~ 61

Author(s):

Elena Carnero ◽

Wenjing Li ◽

Antonio V. Borderia ◽

Bruno Moltedo ◽

Thomas Moran ◽

...

Keyword(s):

Immune Response ◽

Human Immunodeficiency Virus ◽

Immune Responses ◽

Newcastle Disease Virus ◽

Disease Virus ◽

Newcastle Disease ◽

Insertion Site ◽

Gag Protein ◽

Immunodeficiency Virus

ABSTRACT One attractive strategy for the development of a human immunodeficiency virus (HIV) vaccine is the use of viral vectors with a proven safety profile and an absence of preexisting immunity in humans, such as Newcastle disease virus (NDV). Several NDV vaccine vectors have been generated, and their immunogenicities have been investigated with different animal models. However, a systematic study to evaluate the optimal insertion site of the foreign antigens into NDV that results in enhanced immune responses specific to the antigen has not yet been conducted. In this article, we describe the ability of NDV expressing HIV Gag to generate a Gag-specific immune response in mice. We also have determined the optimal insertion site into the NDV genome by generating recombinant NDV-HIVGag viruses in which HIV gag was located at different transcriptional positions throughout the NDV viral genome. All recombinant viruses were viable, grew to similar titers in embryonated chicken eggs, and expressed Gag in a stable manner. Our in vivo experiments revealed that higher HIV Gag protein expression positively correlates with an enhanced CD8+ T-cell-mediated immune response and protective immunity against challenge with vaccinia virus expressing HIV Gag. We also inserted a codon-optimized version of HIV gag in the described best location, between the P and M genes. Virus expressing the codon-optimized version of HIV gag induced a higher expression of the protein and an enhanced immune response against HIV Gag in mice. These results indicate that strategies directed toward increasing antigen expression by NDV result in enhanced immunogenicity and vaccine efficacy.

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Mucosal Immunization with Newcastle Disease Virus Vector Coexpressing HIV-1 Env and Gag Proteins Elicits Potent Serum, Mucosal, and Cellular Immune Responses That Protect against Vaccinia Virus Env and Gag Challenges

mBio ◽

10.1128/mbio.01005-15 ◽

2015 ◽

Vol 6 (4) ◽

Cited By ~ 21

Author(s):

Sunil K. Khattar ◽

Vinoth Manoharan ◽

Bikash Bhattarai ◽

Celia C. LaBranche ◽

David C. Montefiori ◽

...

Keyword(s):

Immune Responses ◽

Newcastle Disease Virus ◽

Disease Virus ◽

Newcastle Disease ◽

Effective Vaccine ◽

Gag Proteins ◽

Humoral Immune ◽

Vaccinia Viruses ◽

Mucosal Immune Responses ◽

Hiv 1

ABSTRACT Newcastle disease virus (NDV) avirulent strain LaSota was used to coexpress gp160 Env and p55 Gag from a single vector to enhance both Env-specific and Gag-specific immune responses. The optimal transcription position for both Env and Gag genes in the NDV genome was determined by generating recombinant NDV (rNDV)-Env-Gag (gp160 located between the P and M genes and Gag between the HN and L genes), rNDV-Gag-Env (Gag located between the P and M genes and gp160 between the HN and L genes), rNDV-Env/Gag (gp160 followed by Gag located between the P and M genes), and rNDV-Gag/Env (Gag followed by gp160 located between the P and M genes). All the recombinant viruses replicated at levels similar to those seen with parental NDV in embryonated chicken eggs and in chicken fibroblast cells. Both gp160 and Gag proteins were expressed at high levels in cell culture, with gp160 found to be incorporated into the envelope of NDV. The Gag and Env proteins expressed by all the recombinants except rNDV-Env-Gag self-assembled into human immunodeficiency virus type 1 (HIV-1) virus-like particles (VLPs). Immunization of guinea pigs by the intranasal route with these rNDVs produced long-lasting Env- and Gag-specific humoral immune responses. The Env-specific humoral and mucosal immune responses and Gag-specific humoral immune responses were higher in rNDV-Gag/Env and rNDV-Env/Gag than in the other recombinants. rNDV-Gag/Env and rNDV-Env/Gag were also more efficient in inducing cellular as well as protective immune responses to challenge with vaccinia viruses expressing HIV-1 Env and Gag in mice. These results suggest that vaccination with a single rNDV coexpressing Env and Gag represents a promising strategy to enhance immunogenicity and protective efficacy against HIV. IMPORTANCE A safe and effective vaccine that can induce both systemic and mucosal immune responses is needed to control HIV-1. In this study, we showed that coexpression of Env and Gag proteins of HIV-1 performed using a single Newcastle disease virus (NDV) vector led to the formation of HIV-1 virus-like particles (VLPs). Immunization of guinea pigs with recombinant NDVs (rNDVs) elicited potent long-lasting systemic and mucosal immune responses to HIV. Additionally, the rNDVs were efficient in inducing cellular immune responses to HIV and protective immunity to challenge with vaccinia viruses expressing HIV Env and Gag in mice. These results suggest that the use of a single NDV expressing Env and Gag proteins simultaneously is a novel strategy to develop a safe and effective vaccine against HIV.

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Evaluation of Newcastle disease virus mediated dendritic cell activation and cross‐priming tumor‐specific immune responses ex vivo

International Journal of Cancer ◽

10.1002/ijc.32694 ◽

2019 ◽

Vol 146 (2) ◽

pp. 531-541 ◽

Cited By ~ 1

Author(s):

Qi Xu ◽

Udaya S. Rangaswamy ◽

Weijia Wang ◽

Scott H. Robbins ◽

James Harper ◽

...

Keyword(s):

Dendritic Cell ◽

Immune Responses ◽

Newcastle Disease Virus ◽

Disease Virus ◽

Newcastle Disease ◽

Cell Activation ◽

Ex Vivo ◽

Dendritic Cell Activation

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Newcastle Disease Virus‐Like Particles: Preparation, Purification, Quantification, and Incorporation of Foreign Glycoproteins

Current Protocols in Microbiology ◽

10.1002/9780471729259.mc1802s30 ◽

2013 ◽

Vol 30 (1) ◽

Cited By ~ 8

Author(s):

Lori W. McGinnes ◽

Trudy G. Morrison

Keyword(s):

Newcastle Disease Virus ◽

Disease Virus ◽

Newcastle Disease ◽

Virus Like Particles

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Requirements for the Assembly and Release of Newcastle Disease Virus-Like Particles

Journal of Virology ◽

10.1128/jvi.00726-06 ◽

2006 ◽

Vol 80 (22) ◽

pp. 11062-11073 ◽

Cited By ~ 104

Author(s):

Homer D. Pantua ◽

Lori W. McGinnes ◽

Mark E. Peeples ◽

Trudy G. Morrison

Keyword(s):

Newcastle Disease Virus ◽

Disease Virus ◽

Newcastle Disease ◽

Plasma Membranes ◽

Matrix Protein ◽

Viral Proteins ◽

M Protein ◽

Virion Assembly ◽

Virus Like Particles ◽

Necessary And Sufficient

ABSTRACT Paramyxoviruses, such as Newcastle disease virus (NDV), assemble in and bud from plasma membranes of infected cells. To explore the role of each of the NDV structural proteins in virion assembly and release, virus-like particles (VLPs) released from avian cells expressing all possible combinations of the nucleoprotein (NP), membrane or matrix protein (M), an uncleaved fusion protein (F-K115Q), and hemagglutinin-neuraminidase (HN) protein were characterized for densities, protein content, and efficiencies of release. Coexpression of all four proteins resulted in the release of VLPs with densities and efficiencies of release (1.18 to 1.16 g/cm3 and 83.8% ± 1.1%, respectively) similar to those of authentic virions. Expression of M protein alone, but not NP, F-K115Q, or HN protein individually, resulted in efficient VLP release, and expression of all different combinations of proteins in the absence of M protein did not result in particle release. Expression of any combination of proteins that included M protein yielded VLPs, although with different densities and efficiencies of release. To address the roles of NP, F, and HN proteins in VLP assembly, the interactions of proteins in VLPs formed with different combinations of viral proteins were characterized by coimmunoprecipitation. The colocalization of M protein with cell surface F and HN proteins in cells expressing all combinations of viral proteins was characterized. Taken together, the results show that M protein is necessary and sufficient for NDV budding. Furthermore, they suggest that M-HN and M-NP interactions are responsible for incorporation of HN and NP proteins into VLPs and that F protein is incorporated indirectly due to interactions with NP and HN protein.

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