Chimeric virus-like particles of universal antigen epitopes of coronavirus and phage Qβ coat protein trigger the production of neutralizing antibodies

Background: Virus-like particles (VLPs) are non-genetic multimeric nanoparticles synthesized through in vitro or in vivo self-assembly of one or more viral structural proteins. Immunogenicity and safety of VLPs make them ideal candidates for vaccine development and efficient nanocarriers for foreign antigens or adjuvants to activate the immune system. Aims: The present study aimed to design and synthesize a chimeric VLP vaccine of the phage Qbeta (Qβ) coat protein presenting the universal epitope of the coronavirus. Methods: The RNA phage Qβ coat protein was designed and synthesized, denoted as Qbeta. The CoV epitope, a universal epitope of coronavirus, was inserted into the C-terminal of Qbeta using genetic recombination, which was designated as Qbeta-CoV. The N-terminal of Qbeta-CoV was successively inserted into the TEV restriction site using mCherry red fluorescent label and modified affinity-purified histidine label 6xHE, which was denoted as HE-Qbeta-CoV. Isopropyl β-D-1-thiogalactopyranoside (IPTG) assessment revealed the expression of Qbeta, Qbeta-CoV, and HE-Qbeta-CoV in the BL21 (DE3) cells. The fusion protein was purified by salting out using ammonium sulfate and affinity chromatography. The morphology of particles was observed using electron microscopy. The female BALB/C mice were immunized intraperitoneally with the Qbeta-CoV and HE-Qbeta-CoV chimeric VLPs vaccines. Their sera were collected for the detection of antibody level and antibody titer using ELISA. The serum is used for the neutralization test of the three viruses of MHV, PEDV, and PDCoV. Results: The results revealed that the fusion proteins Qbeta, Qbeta-CoV, and HE-Qbeta-CoV could all obtain successful expression. Particles with high purity were obtained after purification; the chimeric particles of Qbeta-CoV and HE-Qbeta-CoV were found to be similar to Qbeta particles in morphology and formed chimeric VLPs. In addition, two chimeric VLP vaccines induced specific antibody responses in mice, and the antibodies showed certain neutralizing activity. Conclusion: The successful construction of the chimeric VLPs of the phage Qβ coat protein presenting the universal epitope of coronavirus provides a vaccine form with potential clinical applications for the treatment of coronavirus disease.

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Characteristics of Artificial Virus-like Particles Assembled in vitro from Potato Virus X Coat Protein and Foreign Viral RNAs

Acta Naturae ◽

10.32607/20758251-2011-3-3-40-46 ◽

2011 ◽

Vol 3 (3) ◽

pp. 40-46 ◽

Cited By ~ 8

Author(s):

M V Arkhipenko ◽

E K Petrova ◽

N A Nikitin ◽

A D Protopopova ◽

E V Dubrovin ◽

...

Keyword(s):

Coat Protein ◽

Potato Virus ◽

Potato Virus X ◽

Viral Rnas ◽

Virus Like Particles ◽

Artificial Virus

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Effects of Size and Surface Properties of Nanodiamonds on the Immunogenicity of Plant-Based H5 Protein of A/H5N1 Virus in Mice

Nanomaterials ◽

10.3390/nano11061597 ◽

2021 ◽

Vol 11 (6) ◽

pp. 1597

Author(s):

Thuong Thi Ho ◽

Van Thi Pham ◽

Tra Thi Nguyen ◽

Vy Thai Trinh ◽

Tram Vi ◽

...

Keyword(s):

High Pressure ◽

Neutralizing Antibodies ◽

H5n1 Virus ◽

Vaccine Development ◽

Particle Characterization ◽

Innovative Strategy ◽

Specific Igg ◽

Positively Charged

Nanodiamond (ND) has recently emerged as a potential nanomaterial for nanovaccine development. Here, a plant-based haemagglutinin protein (H5.c2) of A/H5N1 virus was conjugated with detonation NDs (DND) of 3.7 nm in diameter (ND4), and high-pressure and high-temperature (HPHT) oxidative NDs of ~40–70 nm (ND40) and ~100–250 nm (ND100) in diameter. Our results revealed that the surface charge, but not the size of NDs, is crucial to the protein conjugation, as well as the in vitro and in vivo behaviors of H5.c2:ND conjugates. Positively charged ND4 does not effectively form stable conjugates with H5.c2, and has no impact on the immunogenicity of the protein both in vitro and in vivo. In contrast, the negatively oxidized NDs (ND40 and ND100) are excellent protein antigen carriers. When compared to free H5.c2, H5.c2:ND40, and H5.c2:ND100 conjugates are highly immunogenic with hemagglutination titers that are both 16 times higher than that of the free H5.c2 protein. Notably, H5.c2:ND40 and H5.c2:ND100 conjugates induce over 3-folds stronger production of both H5.c2-specific-IgG and neutralizing antibodies against A/H5N1 than free H5.c2 in mice. These findings support the innovative strategy of using negatively oxidized ND particles as novel antigen carriers for vaccine development, while also highlighting the importance of particle characterization before use.

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Conjugation of an antibody Fv fragment to a virus coat protein: cell-specific targeting of recombinant polyoma-virus-like particles

Biochemical Journal ◽

10.1042/bj3560867 ◽

2001 ◽

Vol 356 (3) ◽

pp. 867-873 ◽

Cited By ~ 20

Author(s):

Kay STUBENRAUCH ◽

Stefan GLEITER ◽

Ulrich BRINKMANN ◽

Rainer RUDOLPH ◽

Hauke LILIE

Keyword(s):

Coat Protein ◽

Mammalian Cells ◽

Recombinant Antibody ◽

Specific Antigen ◽

Antibody Fragment ◽

Fusion Peptides ◽

Virus Like Particles ◽

Vector Systems ◽

Virus Coat

The development of cell-type-specific delivery systems is highly desirable for gene-therapeutic applications. Current virus-based vector systems show broad cell specificity, which results in the need to restrict the natural tropism of these viral systems. Here we demonstrate that tumour-cell-specific virus-like particles can be functionally assembled in vitro from recombinant viral coat protein expressed in Escherichia coli. The insertion of a negatively charged peptide in the HI loop of polyoma VP1 interferes with the binding of VP1 to the natural recognition site on mammalian cells and also serves as an adapter for the coupling of antibody fragments that contain complementary charged fusion peptides. A recombinant antibody fragment of the tumour-specific anti-(Lewis Y) antibody B3 could be coupled to the mutant VP1 by engineered polyionic peptides and an additional disulphide bond. With this system an entirely recombinant cell-specific delivery system assembled in vitro could be generated that transfers genes preferentially to cells presenting the tumour-specific antigen on the cell surface.

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A Novel MVA-Based HIV Vaccine Candidate (MVA-gp145-GPN) Co-Expressing Clade C Membrane-Bound Trimeric gp145 Env and Gag-Induced Virus-Like Particles (VLPs) Triggered Broad and Multifunctional HIV-1-Specific T Cell and Antibody Responses

Viruses ◽

10.3390/v11020160 ◽

2019 ◽

Vol 11 (2) ◽

pp. 160 ◽

Cited By ~ 4

Author(s):

Beatriz Perdiguero ◽

Cristina Sánchez-Corzo ◽

Carlos Sorzano ◽

Lidia Saiz ◽

Pilar Mediavilla ◽

...

Keyword(s):

Neutralizing Antibodies ◽

Vaccine Candidate ◽

Hiv Vaccine ◽

Replication Capacity ◽

Virus Like Particles ◽

Modified Vaccinia Virus Ankara ◽

Clade C ◽

Membrane Bound ◽

Hiv 1

The development of an effective Human Immunodeficiency Virus (HIV) vaccine that is able to stimulate both the humoral and cellular HIV-1-specific immune responses remains a major priority challenge. In this study, we described the generation and preclinical evaluation of single and double modified vaccinia virus Ankara (MVA)-based candidates expressing the HIV-1 clade C membrane-bound gp145(ZM96) trimeric protein and/or the Gag(ZM96)-Pol-Nef(CN54) (GPN) polyprotein that was processed to form Gag-induced virus-like particles (VLPs). In vitro characterization of MVA recombinants revealed the stable integration of HIV-1 genes without affecting its replication capacity. In cells that were infected with Env-expressing viruses, the gp145 protein was inserted into the plasma membrane exposing critical epitopes that were recognized by broadly neutralizing antibodies (bNAbs), whereas Gag-induced VLPs were released from cells that were infected with GPN-expressing viruses. VLP particles as well as purified MVA virions contain Env and Gag visualized by immunoelectron microscopy and western-blot of fractions that were obtained after detergent treatments of purified virus particles. In BALB/c mice, homologous MVA-gp145-GPN prime/boost regimen induced broad and polyfunctional Env- and Gag-specific CD4 T cells and antigen-specific T follicular helper (Tfh) and Germinal Center (GC) B cells, which correlated with robust HIV-1-specific humoral responses. Overall, these results support the consideration of MVA-gp145-GPN vector as a potential vaccine candidate against HIV-1.

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Mucosal but Not Parenteral Immunization with Purified Human Papillomavirus Type 16 Virus-Like Particles Induces Neutralizing Titers of Antibodies throughout the Estrous Cycle of Mice

Journal of Virology ◽

10.1128/jvi.73.11.9609-9613.1999 ◽

1999 ◽

Vol 73 (11) ◽

pp. 9609-9613 ◽

Cited By ~ 57

Author(s):

Denise Nardelli-Haefliger ◽

Richard Roden ◽

Carole Balmelli ◽

Alexandra Potts ◽

John Schiller ◽

...

Keyword(s):

Human Papillomavirus ◽

Estrous Cycle ◽

Neutralizing Antibodies ◽

Female Genital Tract ◽

Neutralization Assay ◽

Enzyme Linked Immunosorbent Assay ◽

Virus Like Particles ◽

Human Papillomavirus Type 16 ◽

Parenteral Immunization

ABSTRACT We have recently shown that nasal immunization of anesthetized mice with human papillomavirus type 16 (HPV16) virus-like particles (VLPs) is highly effective at inducing both neutralizing immunoglobulin A (IgA) and IgG in genital secretions, while parenteral immunization induced only neutralizing IgG. Our data also demonstrated that both isotypes are similarly neutralizing according to an in vitro pseudotyped neutralization assay. However, it is known that various amounts of IgA and IgG are produced in genital secretions along the estrous cycle. Therefore, we have investigated how this variation influences the amount of HPV16 neutralizing antibodies induced after immunization with VLPs. We have compared parenteral and nasal protocols of vaccination with daily samplings of genital secretions of mice. Enzyme-linked immunosorbent assay analysis showed that total IgA and IgG inversely varied along the estrous cycle, with the largest amounts of IgA in proestrus-estrus and the largest amount of IgG in diestrus. This resulted in HPV16 neutralizing titers of IgG only being achieved during diestrus upon parenteral immunization. In contrast, nasal vaccination induced neutralizing titers of IgA plus IgG throughout the estrous cycle, as confirmed by in vitro pseudotyped neutralization assays. Our data suggest that mucosal immunization might be more efficient than parenteral immunization at inducing continuous protection of the female genital tract.

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Cellular and Humoral Immunogenicity of a Candidate DNA Vaccine Expressing SARS-CoV-2 Spike Subunit 1

Vaccines ◽

10.3390/vaccines9080852 ◽

2021 ◽

Vol 9 (8) ◽

pp. 852

Author(s):

Khalid A. Alluhaybi ◽

Rahaf H. Alharbi ◽

Rowa Y. Alhabbab ◽

Najwa D. Aljehani ◽

Sawsan S. Alamri ◽

...

Keyword(s):

Immune Responses ◽

Dna Vaccine ◽

Neutralizing Antibodies ◽

Vaccine Development ◽

Cell Responses ◽

Cellular Immune Responses ◽

Specific Igg ◽

Cellular Immune ◽

Different Doses

The urgent need for effective, safe and equitably accessible vaccines to tackle the ongoing spread of COVID-19 led researchers to generate vaccine candidates targeting varieties of immunogens of SARS-CoV-2. Because of its crucial role in mediating binding and entry to host cell and its proven safety profile, the subunit 1 (S1) of the spike protein represents an attractive immunogen for vaccine development. Here, we developed and assessed the immunogenicity of a DNA vaccine encoding the SARS-CoV-2 S1. Following in vitro confirmation and characterization, the humoral and cellular immune responses of our vaccine candidate (pVAX-S1) was evaluated in BALB/c mice using two different doses, 25 µg and 50 µg. Our data showed high levels of SARS-CoV-2 specific IgG and neutralizing antibodies in mice immunized with three doses of pVAX-S1. Analysis of the induced IgG subclasses showed a Th1-polarized immune response, as demonstrated by the significant elevation of spike-specific IgG2a and IgG2b, compared to IgG1. Furthermore, we found that the immunization of mice with three doses of 50 µg of pVAX-S1 could elicit significant memory CD4+ and CD8+ T cell responses. Taken together, our data indicate that pVAX-S1 is immunogenic and safe in mice and is worthy of further preclinical and clinical evaluation.

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Current Overviews on COVID-19 Management Strategies

Current Pharmaceutical Biotechnology ◽

10.2174/1389201022666210509022313 ◽

2021 ◽

Vol 22 ◽

Author(s):

Pankaj Kumar Rai ◽

Zeba Mueed ◽

Abhiroop Chowdhury ◽

Ravi Deval ◽

Dinesh Kumar ◽

...

Keyword(s):

Neutralizing Antibodies ◽

Vaccine Development ◽

Management Strategies ◽

Symptomatic Treatment ◽

Diagnostic Methods ◽

Antibody Detection ◽

Modes Of Action ◽

Prospective Control ◽

Wide Range

The coronavirus pandemic has hit the world lately and caused acute respiratory syndrome in humans. The causative agent of the disease was soon brought to focus by scientists as SARS-CoV-2 and later called a novel coronavirus by the general public. Due to the severity and rapid spread of the disease, WHO classifies the COVID-19 pandemic as the 6th public health emergency even after taking efforts like worldwide quarantine and restrictions. Since only symptomatic treatment is available, the best way to control the spread of the virus is by taking preventive measures. Various types of antigen/antibody detection kits and diagnostic methods are available for the diagnosis of COVID-19 patients. In recent years, various phytochemicals and repurposing drugs are showing a broad range of anti-viral activities with different modes of action have been identified. Repurposing drugs such as arbidol, hydroxychloroquine, chloroquine, lopinavir, favipiravir, remdesivir, hexamethylene amiloride, and dexamethasone, tocilizumab, interferon-β, neutralizing antibodies exhibit in vitro anti-coronaviral properties by inhibiting multiple processes in the virus life cycle. Various research groups are involved in drug trials and vaccine development. Plant-based anti-viral compounds such as baicalin, calanolides, curcumin, oxymatrine, matrine, and resveratrol exhibit different modes of action against a wide range of positive/negative sense-RNA/DNA virus, and future researches need to be conducted to ascertain their role, use in managing SARS-CoV-2. Thus, this article is an attempt to review the current understanding of COVID-19 acute respiratory disease and summarize its clinical features with their prospective control and various aspects of the therapeutic approach.

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Generation of Neutralizing Human Monoclonal Antibodies against Parvovirus B19 Proteins

Journal of Virology ◽

10.1128/jvi.73.3.1974-1979.1999 ◽

1999 ◽

Vol 73 (3) ◽

pp. 1974-1979 ◽

Cited By ~ 103

Author(s):

Andreas Gigler ◽

Simone Dorsch ◽

Andrea Hemauer ◽

Constance Williams ◽

Sonnie Kim ◽

...

Keyword(s):

Monoclonal Antibodies ◽

Capsid Protein ◽

Neutralizing Antibodies ◽

Vaccine Development ◽

Nonstructural Protein ◽

Parvovirus B19 ◽

Human Mabs ◽

Unique Region ◽

Neutralizing Activity

ABSTRACT Infections caused by human parvovirus B19 are known to be controlled mainly by neutralizing antibodies. To analyze the immune reaction against parvovirus B19 proteins, four cell lines secreting human immunoglobulin G monoclonal antibodies (MAbs) were generated from two healthy donors and one human immunodeficiency virus type 1-seropositive individual with high serum titers against parvovirus. One MAb is specific for nonstructural protein NS1 (MAb 1424), two MAbs are specific for the unique region of minor capsid protein VP1 (MAbs 1418-1 and 1418-16), and one MAb is directed to major capsid protein VP2 (MAb 860-55D). Two MAbs, 1418-1 and 1418-16, which were generated from the same individual have identity in the cDNA sequences encoding the variable domains, with the exception of four base pairs resulting in only one amino acid change in the light chain. The NS1- and VP1-specific MAbs interact with linear epitopes, whereas the recognized epitope in VP2 is conformational. The MAbs specific for the structural proteins display strong virus-neutralizing activity. The VP1- and VP2-specific MAbs have the capacity to neutralize 50% of infectious parvovirus B19 in vitro at 0.08 and 0.73 μg/ml, respectively, demonstrating the importance of such antibodies in the clearance of B19 viremia. The NS1-specific MAb mediated weak neutralizing activity and required 47.7 μg/ml for 50% neutralization. The human MAbs with potent neutralizing activity could be used for immunotherapy of chronically B19 virus-infected individuals and acutely infected pregnant women. Furthermore, the knowledge gained regarding epitopes which induce strongly neutralizing antibodies may be important for vaccine development.

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Preparation of functional IgY that potently neutralizes HIV-1 in TZM-bl Cell line

10.1101/2021.03.04.431083 ◽

2021 ◽

Author(s):

Jinchen Wei ◽

Yanqun Zhang ◽

Yonglian Zeng ◽

Yang Yang ◽

Ronggan Liang ◽

...

Keyword(s):

Cell Line ◽

Neutralizing Antibodies ◽

Evaluation System ◽

Salting Out ◽

Novel Technology ◽

Public Health Challenges ◽

Hiv Neutralization ◽

Hiv 1

ABSTRACTAIDS caused by HIV is one of the most serious public health challenges in the world. As we all know, Antiretroviral therapy (ART) is the most effective way to treat AIDS so far, however, forthe reasons of drug resistance, side effects, compliance, economy, limited its using widely. On the other hand, AIDS cannot be completely cured by ART. While the characterization of bnAbs (broadly neutralizing antibodies) in potent HIV neutralization provides considerable insight into HIV curing, it also can be used for passive immunotherapy or combination with ART for HIV-1 treatment. Here we report a novel technology to produce an neutralized activity bnAbs named HIV-1-IgY, which was extracted from the immunized Chicken egg by pNL4-3 virus antigens, and further purified using Water dilution and Salting out method. The specificity, titer and neutralizing activity of HIV-1-IgY was analyzed by Western blotting, ELISA and TZM-bl cell line evaluation system respectively. The results showed that theHIV-1-IgY has high neutralized activity to HIV in vitro; nearly 90% of HIV-1 were neutralized at 1.89μM in TZM-blsystem, which indicated that IgY may be a source of antibodies for AIDS prevention and treatment. Despite its needs to further consider and evaluate neutralized activity in-vivo and the potential mechanisms, Our data showed that we obtained an HIV-1-IgY which could effectively neutralize HIV-l IIIB virus in vitro.

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A Novel Recombinant Virus-Like Particles Displaying B and T Cell Epitopes of Japanese Encephalitis Virus Offers Protective Immunity in Mice and Guinea Pigs

Vaccines ◽

10.3390/vaccines9090980 ◽

2021 ◽

Vol 9 (9) ◽

pp. 980

Author(s):

Muhammad Naveed Anwar ◽

Chunying Jiang ◽

Di Di ◽

Junjie Zhang ◽

Shuang Guo ◽

...

Keyword(s):

Japanese Encephalitis Virus ◽

Japanese Encephalitis ◽

Guinea Pigs ◽

Neutralizing Antibodies ◽

Protective Immunity ◽

Vaccine Development ◽

Amino Acid Sequences ◽

Encephalitis Virus ◽

Potential Candidate ◽

Virus Like Particles

Virus-like particles (VLPs) are non-replicative vectors for the delivery of heterologous epitopes and are considered one of the most potent inducers of cellular and humoral immune responses in mice and guinea pigs. In the present study, VLP-JEVe was constructed by the insertion of six Japanese encephalitis virus (JEV) envelope protein epitopes into different surface loop regions of PPV VP2 by the substitution of specific amino acid sequences without altering the assembly of the virus; subsequently, the protective efficacy of this VLP-JEVe was evaluated against JEV challenge in mice and guinea pigs. Mice immunized with the VLP-JEVe antigen developed high titers of neutralizing antibodies and 100% protection against lethal JEV challenge. The neutralizing and hemagglutination inhibition (HI) antibody responses were also induced in guinea pigs vaccinated with VLP-JEVe. In addition, immunization with VLP-JEVe in mice induced effective neutralizing antibodies and protective immunity against PPV (porcine parvovirus) challenge in guinea pigs. These studies suggest that VLP-JEVe produced as described here could be a potential candidate for vaccine development.

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