scholarly journals Recombinant Adenovirus Expressing Vesicular Stomatitis Virus G Proteins induce both humoral and cell-medicated immune responses in Mice and Goats

2020 ◽  
Author(s):  
Xiaojuan Xue ◽  
Zhaorong Yu ◽  
Hongyan Jin ◽  
Lin Liang ◽  
Jiayang Li ◽  
...  
Keyword(s):  
Immune Responses ◽  
G Proteins ◽  
Vesicular Stomatitis Virus ◽  
Immune Cell ◽  
Recombinant Adenovirus ◽  
Negative Control ◽  
Direct Immunofluorescence ◽  
Post Inoculation ◽  
293 Cells ◽  
Vesicular Stomatitis

Abstract Background: With a human type 5 replication-defective adenovirus expression vector, we constructed the three recombinant adenoviruses (rAd) and expressed the Vesicular Stomatitis Virus (VSV) Indiana serotype glycoprotein (VSV-IN-G), VSV New Jersey serotype glycoprotein (VSV-NJ-G), and the G fusion protein [two serotypes G (VSV-IN-G-NJ-G)]. Three rAds were named rAd-IN, rAd-NJ, and rAd-IN-NJ. The three rAds were inoculated into AAV-293 cells, and the AAV-293 cells were serially propagated to 20 generations until the virus titers were stable, then TCID50 was determined. In direct immunofluorescence and western blot were used for detecting the expression of the target proteins and lymphocyte proliferation test was used for immune cell numbers. Results: The results showed that G proteins we expressed with good reactogenicity. The rAds were used to subcutaneously inoculate mice three times with 2-week intervals, and goats two times with 3-week intervals, respectively. On 0, 2, 4, and 6 weeks of post-inoculation for the mice and 0, 3, 6, 9, and 12 weeks for goats, their sera were collected and NT antibodies were determined. The results showed that the rAds could induce the production of VSV antibodies in the mice, and VSV NT antibodies in the goats. The antibody levels were 1:16 to 1: 32 in mice, and 1:32 to 1: 64 in the goats. The rAds induced strong immune lymphocyte proliferations in mice and goats, which was significantly higher than those of the negative control groups. Conclusion: The three rAds expressed VSV-G proteins at high levels, and induced humoral and cellular immune responses in both mice and goats, which laid a foundation for further studies of the recombinant adenovirus vaccines expressing VSV glycoprotein.

scholarly journals Recombinant adenovirus expressing vesicular stomatitis virus G proteins induce both humoral and cell-mediated immune responses in mice and goats

2021 ◽  
Vol 17 (1) ◽  
Author(s):  
Xiaojuan Xue ◽  
Zhaorong Yu ◽  
Hongyan Jin ◽  
Lin Liang ◽  
Jiayang Li ◽  
...  
Keyword(s):  
Immune Responses ◽  
G Proteins ◽  
Vesicular Stomatitis Virus ◽  
Neutralizing Antibodies ◽  
Recombinant Adenovirus ◽  
Neutralizing Antibody ◽  
Negative Control ◽  
Antibody Titers ◽  
Vesicular Stomatitis ◽  
Cellular Immune

Abstract Background Vesicular stomatitis (VS) is an acute, highly contagious and economically important zoonotic disease caused by the vesicular stomatitis virus (VSV). There is a need for effective and safe stable recombinant vaccine for the control of the disease. The human type 5 replication-defective adenovirus expression vector is a good way to construct recombinant vaccines. Results Three recombinant adenoviruses (rAd) were successfully constructed that expressed the VSV Indiana serotype glycoprotein (VSV-IN-G), VSV New Jersey serotype glycoprotein (VSV-NJ-G), and the G fusion protein (both serotypes of G [VSV-IN-G-NJ-G]) with potentiality to induce protective immunity. G proteins were successfully expressed with good immunogenicity. The rAds could induce the production of VSV antibodies in mice, and VSV neutralizing antibodies in goats, respectively. The neutralizing antibody titers could reach 1:32 in mice and 1:64 in goats. The rAds induced strong lymphocyte proliferation in mice and goats, which was significantly higher compared to the negative control groups. Conclusions The three rAds constructed in the study expressed VSV-G proteins and induced both humoral and cellular immune responses in mice and goats. These results lay the foundation for further studies on the use of rAds in vaccines expressing VSV-G.

scholarly journals Recombinant Adenovirus Expressing Vesicular Stomatitis Virus G Proteins Induce both Humoral and Cell-mediated Immune Responses in Mice and Goats

2021 ◽  
Author(s):  
Xiaojuan Xue ◽  
Zhaorong Yu ◽  
Hongyan Jin ◽  
Lin Liang ◽  
Jiayang Li ◽  
...  
Keyword(s):  
Immune Responses ◽  
G Proteins ◽  
Vesicular Stomatitis Virus ◽  
Neutralizing Antibodies ◽  
Recombinant Adenovirus ◽  
Neutralizing Antibody ◽  
Negative Control ◽  
Antibody Titers ◽  
Vesicular Stomatitis ◽  
Cellular Immune

Abstract Background: Vesicular stomatitis (VS) is an acute, highly contagious and economically important zoonotic disease caused by the vesicular stomatitis virus (VSV). There is a need for effective and safe stable recombinant vaccine for the control of the disease. The human type 5 replication-defective adenovirus expression vector is a good way to construct recombinant vaccines.Results: Three recombinant adenoviruses (rAd) were successfully constructed that expressed the VSV Indiana serotype glycoprotein (VSV-IN-G), VSV New Jersey serotype glycoprotein (VSV-NJ-G), and the G fusion protein (both serotypes of G [VSV-IN-G-NJ-G]) with potentiality to induce protective immunity. G proteins were successfully expressed with good immunogenicity. The rAds could induce the production of VSV antibodies in mice, and VSV neutralizing antibodies in goats, respectively. The neutralizing antibody titers could reach 1:32 in mice and 1:64 in goats. The rAds induced strong lymphocyte proliferation in mice and goats, which was significantly higher compared to the negative control groups. Conclusions: The three rAds constructed in the study expressed VSV-G proteins and induced both humoral and cellular immune responses in mice and goats. These results lay the foundation for further studies on the use of rAds in vaccines expressing VSV-G.

scholarly journals Characterisation of Antibody Interactions with the G Protein of Vesicular Stomatitis Virus Indiana Strain and Other Vesiculovirus G Proteins

2018 ◽  
Author(s):  
Altar M Munis ◽  
Maha Tijani ◽  
Mark Hassall ◽  
Giada Mattiuzzo ◽  
Mary K Collins ◽  
...  
Keyword(s):  
G Protein ◽  
G Proteins ◽  
Vesicular Stomatitis Virus ◽  
Conformational Changes ◽  
Lentiviral Vectors ◽  
Medicinal Products ◽  
Vaccine Vectors ◽  
Receptor Binding Site ◽  
Advanced Therapy ◽  
Vesicular Stomatitis

ABSTRACTVesicular stomatitis virus Indiana strain G protein (VSVind.G) is the most commonly used envelope glycoprotein to pseudotype lentiviral vectors (LV) for experimental and clinical applications. Recently, G proteins derived from other vesiculoviruses (VesG), for example Cocal virus, have been proposed as alternative LV envelopes with possible advantages compared to VSVind.G. Well-characterised antibodies that recognise VesG will be useful for vesiculovirus research, development of G protein-containing advanced therapy medicinal products (ATMPs), and deployment of VSVind-based vaccine vectors. Here we show that one commercially available monoclonal antibody, 8G5F11, binds to and neutralises G proteins from three strains of VSV as well as Cocal, and Maraba viruses, whereas the other commercially available monoclonal anti-VSVind.G antibody, IE9F9, binds to and neutralises only VSVind.G. Using a combination of G protein chimeras and site-directed mutations, we mapped the binding epitopes of IE9F9 and 8G5F11 on VSVind.G. IE9F9 binds close to the receptor binding site and competes with soluble low-density lipoprotein receptor (LDLR) for binding to VSVind.G, explaining its mechanism of neutralisation. In contrast, 8G5F11 binds close to a region known to undergo conformational changes when the G protein moves to its post-fusion structure, and we propose that 8G5F11 cross-neutralises VesGs by inhibiting this.IMPORTANCEVSVind.G is currently regarded as the gold-standard envelope to pseudotype lentiviral vectors. However, recently other G proteins derived from vesiculoviruses have been proposed as alternative envelopes. Here, we investigated two anti-VSVind.G monoclonal antibodies for their ability to cross-react with other vesiculovirus G proteins, and identified the epitopes they recognise, and explored the mechanisms behind their neutralisation activity. Understanding how cross-neutralising antibodies interact with other G proteins may be of interest in the context of host-pathogen interaction and co-evolution as well as providing the opportunity to modify the G proteins and improve G protein-containing medicinal products and vaccine vectors.

scholarly journals Recombinant Vesicular Stomatitis Virus Vectors Expressing Herpes Simplex Virus Type 2 gD Elicit Robust CD4+ Th1 Immune Responses and Are Protective in Mouse and Guinea Pig Models of Vaginal Challenge

2006 ◽  
Vol 80 (9) ◽  
pp. 4447-4457 ◽  
Author(s):  
Robert J. Natuk ◽  
David Cooper ◽  
Min Guo ◽  
Priscilla Calderon ◽  
Kevin J. Wright ◽  
...  
Keyword(s):  
Herpes Simplex ◽  
Immune Responses ◽  
Vesicular Stomatitis Virus ◽  
Guinea Pigs ◽  
Wild Type ◽  
Humoral Immune ◽  
Humoral Immune Responses ◽  
Vesicular Stomatitis ◽  
Simplex Virus

ABSTRACT Recombinant vesicular stomatitis virus (rVSV) vectors offer an attractive approach for the induction of robust cellular and humoral immune responses directed against human pathogen target antigens. We evaluated rVSV vectors expressing full-length glycoprotein D (gD) from herpes simplex virus type 2 (HSV-2) in mice and guinea pigs for immunogenicity and protective efficacy against genital challenge with wild-type HSV-2. Robust Th1-polarized anti-gD immune responses were demonstrated in the murine model as measured by induction of gD-specific cytotoxic T lymphocytes and increased gamma interferon expression. The isotype makeup of the serum anti-gD immunoglobulin G (IgG) response was consistent with the presence of a Th1-CD4+ anti-gD response, characterized by a high IgG2a/IgG1 IgG subclass ratio. Functional anti-HSV-2 neutralizing serum antibody responses were readily demonstrated in both guinea pigs and mice that had been immunized with rVSV-gD vaccines. Furthermore, guinea pigs and mice were prophylactically protected from genital challenge with high doses of wild-type HSV-2. In addition, guinea pigs were highly protected against the establishment of latent infection as evidenced by low or absent HSV-2 genome copies in dorsal root ganglia after virus challenge. In summary, rVSV-gD vectors were successfully used to elicit potent anti-gD Th1-like cellular and humoral immune responses that were protective against HSV-2 disease in guinea pigs and mice.

scholarly journals H-2Kk and vesicular stomatitis virus G proteins are not extensively associated in reconstituted membranes recognized by T cells.

1982 ◽  
Vol 79 (5) ◽  
pp. 1506-1510 ◽  
Author(s):  
G. S. Cartwright ◽  
L. M. Smith ◽  
E. W. Heinzelmann ◽  
M. J. Ruebush ◽  
J. W. Parce ◽  
...  
Keyword(s):  
T Cells ◽  
G Proteins ◽  
Vesicular Stomatitis Virus ◽  
Vesicular Stomatitis ◽  
Reconstituted Membranes

scholarly journals A methyltransferase-defective VSV-based SARS-CoV-2 vaccine candidate provides complete protection against SARS-CoV-2 infection in hamsters

2021 ◽  
Author(s):  
Mijia Lu ◽  
Yuexiu Zhang ◽  
Piyush Dravid ◽  
Anzhong Li ◽  
Cong Zeng ◽  
...  
Keyword(s):  
Cell Culture ◽  
Immune Responses ◽  
Vesicular Stomatitis Virus ◽  
Vaccine Candidate ◽  
Viral Mrna ◽  
S Protein ◽  
Vaccine Candidates ◽  
Syrian Golden Hamsters ◽  
Vesicular Stomatitis ◽  
Immunocompromised Mice

The current pandemic of coronavirus disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has led to dramatic economic and health burdens. Although the worldwide SARS-CoV-2 vaccination campaign has begun, exploration of other vaccine candidates is needed due to the uncertainties of the current approved vaccines such as durability of protection, cross-protection against variant strains, and costs of long-term production, and storage. In this study, we developed a methyltransferase-defective recombinant vesicular stomatitis virus (mtdVSV)-based SARS-CoV-2 vaccine candidate. We generated mtdVSVs expressing SARS-CoV-2 full-length spike (S), S1, or its receptor binding domain (RBD). All these recombinant viruses grew to high titers in mammalian cells despite high attenuation in cell culture. SARS-CoV-2 S protein and its truncations were highly expressed by the mtdVSV vector. These mtdVSV-based vaccine candidates were completely attenuated in both immunocompetent and immunocompromised mice. Among these constructs, mtdVSV-S induced high levels of SARS-CoV-2 specific neutralizing antibodies (NAbs) and Th1-biased T cell immune responses in mice. Syrian golden hamsters immunized with mtdVSV-S triggered SARS-CoV-2 specific NAbs that were higher than convalescent plasma from convalescent COVID-19 patients. In addition, hamsters immunized with mtdVSV-S were completely protected against SARS-CoV-2 replication in lung and nasal turbinate tissues, cytokine storm, and lung pathology. Collectively, our data demonstrate that mtdVSV expressing SARS-CoV-2 S protein is a safe and highly efficacious vaccine candidate against SARS-CoV-2 infection. Significance Viral mRNA cap methyltransferase (MTase) is essential for mRNA stability, protein translation, and innate immune evasion. Thus, viral mRNA cap MTase activity is a novel target for development of live attenuated or live vectored vaccine candidates. Here, we developed a panel of MTase-defective recombinant recombinant vesicular stomatitis virus (mtdVSV)-based SARS-CoV-2 vaccine candidates expressing full-length S, S1, or several versions of the RBD. These mtdVSV-based vaccine candidates grew to high titers in cell culture and were completely attenuated in both immunocompetent and immunocompromised mice. Among these vaccine candidates, mtdVSV-S induces high levels of SARS-CoV-2 specific neutralizing antibody (Nabs) and Th1-biased immune responses in mice. Syrian golden hamsters immunized with mtdVSV-S triggered SARS-CoV-2 specific NAbs that were higher than convalescent plasma from COVID-19 recovered patients. Furthermore, hamsters immunized with mtdVSV-S were completely protected against SARS-CoV-2 challenge. Thus, mtdVSV is a safe and highly effective vector to deliver SARS-CoV-2 vaccine.

scholarly journals Posttranslational folding of vesicular stomatitis virus G protein in the ER: involvement of noncovalent and covalent complexes.

1993 ◽  
Vol 120 (3) ◽  
pp. 647-655 ◽  
Author(s):  
A de Silva ◽  
I Braakman ◽  
A Helenius
Keyword(s):  
G Protein ◽  
G Proteins ◽  
Vesicular Stomatitis Virus ◽  
Chain Termination ◽  
Metabolic Energy ◽  
Folding Pathway ◽  
Protein Subunits ◽  
Late Step ◽  
Vesicular Stomatitis ◽  
Transient Intermediates

In this study, we show that posttranslational folding of Vesicular Stomatitis virus G protein subunits can involve noncovalent, multimeric complexes as transient intermediates. The complexes are heterogeneous in size (4-21S20,W), contain several G glycopolypeptides, and are associated with BiP/GRP78. The newly synthesized, partially intrachain disulfide-bonded G proteins enter these complexes immediately after chain termination, and are released 1-4 min later as fully oxidized, trimerization-competent monomers. These monomers are properly folded, judging by their binding of conformation-specific mAbs. When the G protein is translated in the presence of DTT, it remains reduced, largely unfolded and aggregated in the ER, but it can fold successfully when the DTT is removed. In this case, contrary to normal folding, the aggregates become transiently disulfide cross-linked. We also demonstrated that the fidelity of the folding process is dependent on metabolic energy. Finally, we established that the G protein of the folding mutant of the Vesicular Stomatitis virus, ts045, is blocked at a relatively late step in the folding pathway and remains associated with oligomeric, BiP/GRP78-containing folding complexes.

Induction of protective immune responses against a lethal Zika virus challenge post-vaccination with a dual serotype of recombinant vesicular stomatitis virus carrying the genetically modified Zika virus E protein gene

2021 ◽  
Vol 102 (4) ◽  
Author(s):  
Jung Ah Choi ◽  
Kunyu Wu ◽  
Gyoung Nyoun Kim ◽  
Nasrin Saeedian ◽  
Seung Han Seon ◽  
...  
Keyword(s):  
Immune Responses ◽  
Vesicular Stomatitis Virus ◽  
Zika Virus ◽  
Protein Gene ◽  
Transmembrane Domain ◽  
Genetically Modified ◽  
Lethal Dose ◽  
Effective Vaccine ◽  
E Protein ◽  
Vesicular Stomatitis

The development of a vaccine to prevent Zika virus (ZIKV) infection has been one of the priorities in infectious disease research in recent years. There have been numerous attempts to develop an effective vaccine against ZIKV. It is imperative to choose the safest and the most effective ZIKV vaccine from all candidate vaccines to control this infection globally. We have employed a dual serotype of prime-boost recombinant vesicular stomatitis virus (VSV) vaccine strategy, to develop a ZIKV vaccine candidate, using a type 1 IFN-receptor knock-out (Ifnar −/−) mouse model for challenge studies. Prime vaccination with an attenuated recombinant VSV Indiana serotype (rVSVInd) carrying a genetically modified ZIKV envelope (E) protein gene followed by boost vaccination with attenuated recombinant VSV New Jersey serotype (rVSVNJ) carrying the same E gene induced robust adaptive immune responses. In particular, rVSV carrying the ZIKV E gene with the honeybee melittin signal peptide (msp) at the N terminus and VSV G protein transmembrane domain and cytoplasmic tail (Gtc) at the C terminus of the E gene induced strong protective immune responses. This vaccine regimen induced highly potent neutralizing antibodies and T cell responses in the absence of an adjuvant and protected Ifnar -/- mice from a lethal dose of the ZIKV challenge.

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