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  1. Article ; Online: Construction of a recombinant avipoxvirus expressing the env gene of Zika virus as a novel putative preventive vaccine.

    Zanotto, Carlo / Paolini, Francesca / Radaelli, Antonia / De Giuli Morghen, Carlo

    Virology journal

    2021  Volume 18, Issue 1, Page(s) 50

    Abstract: Background: Zika virus (ZIKV) has been declared a public health emergency that requires development of an effective vaccine, as it might represent an international threat.: Methods: Here, two novel DNA-based (pVAXzenv) and fowlpox-based (FPzenv) ... ...

    Abstract Background: Zika virus (ZIKV) has been declared a public health emergency that requires development of an effective vaccine, as it might represent an international threat.
    Methods: Here, two novel DNA-based (pVAXzenv) and fowlpox-based (FPzenv) recombinant putative vaccine candidates were constructed that contained the cPrME genes of ZIKV. The env gene inserted into the fowlpox vector was verified for correct transgene expression by Western blotting and by immunofluorescence in different cell lines. The production of virus-like particles as a result of env gene expression was also demonstrated by electron microscopy. BALB/c mice were immunosuppressed with dexamethasone and immunized following a prime-boost strategy in a heterologous protocol where pVAXzenv was followed by FPzenv, to evaluate the immunogenicity of the Env protein. The mice underwent a challenge with an epidemic ZIKV after the last boost.
    Results: These data show that the ZIKV Env protein was correctly expressed in both normal human lung fibroblasts (MRC-5 cells) and green monkey kidney (Vero) cells infected with FPzenv, and that the transgene expression lasted for more than 2 weeks. After mucosal administration of FPzenv, the immunized mice showed specific and significantly higher humoral responses compared to the control mice. However, virus neutralizing antibodies were not detected using plaque reduction assays.
    Conclusions: Although BALB/c mice appear to be an adequate model for ZIKV infection, as it mimics the natural mild infection in human beings, inadequate immune suppression seemed to occur by dexamethasone and different immune suppression strategies should be applied before challenge to reveal any protection of the mice.
    MeSH term(s) Animals ; Antibodies, Neutralizing ; Antibodies, Viral ; Avipoxvirus ; Chlorocebus aethiops ; Dexamethasone ; Fibroblasts ; Gene Products, env ; Genes, env ; Humans ; Immunity, Humoral ; Mice ; Mice, Inbred BALB C ; Vaccines, Synthetic/genetics ; Vero Cells ; Viral Vaccines/genetics ; Zika Virus/genetics ; Zika Virus Infection/prevention & control
    Chemical Substances Antibodies, Neutralizing ; Antibodies, Viral ; Gene Products, env ; Vaccines, Synthetic ; Viral Vaccines ; Dexamethasone (7S5I7G3JQL)
    Language English
    Publishing date 2021-03-04
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2160640-7
    ISSN 1743-422X ; 1743-422X
    ISSN (online) 1743-422X
    ISSN 1743-422X
    DOI 10.1186/s12985-021-01519-x
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article ; Online: Rapid detection of vaccinia virus using biofunctionalized fiber-optic ball-tip biosensors.

    Rakhimbekova, Aida / Kudaibergenov, Baizak / Seitkamal, Kuanysh / Bellone, Aurora / Dauletova, Ayazhan / Sypabekova, Marzhan / Olivero, Massimo / Perrone, Guido / Radaelli, Antonia / Zanotto, Carlo / De Giuli Morghen, Carlo / Vangelista, Luca / Tosi, Daniele

    Scientific reports

    2023  Volume 13, Issue 1, Page(s) 17470

    Abstract: In this work, we present the development and biofunctionalization of a fiber-optic ball-resonator biosensor for the real-time detection of vaccinia poxvirus. We fabricated several ball-tip resonators, functionalized through a silanization process to ... ...

    Abstract In this work, we present the development and biofunctionalization of a fiber-optic ball-resonator biosensor for the real-time detection of vaccinia poxvirus. We fabricated several ball-tip resonators, functionalized through a silanization process to immobilize two bioreceptors: the monoclonal anti-L1R antibody targeting the L1R protein, and the polyclonal rabbit serum antibodies targeting the whole vaccinia virus (VV) pathogen. Experimental measurements were carried out to detect VV in concentrations from 10
    MeSH term(s) Animals ; Humans ; Rabbits ; Vaccinia virus ; Vaccinia ; Poxviridae ; Fiber Optic Technology ; Biosensing Techniques
    Language English
    Publishing date 2023-10-14
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2615211-3
    ISSN 2045-2322 ; 2045-2322
    ISSN (online) 2045-2322
    ISSN 2045-2322
    DOI 10.1038/s41598-023-44926-6
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  3. Article: Computational Design and Development of Benzodioxane-Benzamides as Potent Inhibitors of FtsZ by Exploring the Hydrophobic Subpocket.

    Straniero, Valentina / Sebastián-Pérez, Victor / Suigo, Lorenzo / Margolin, William / Casiraghi, Andrea / Hrast, Martina / Zanotto, Carlo / Zdovc, Irena / Radaelli, Antonia / Valoti, Ermanno

    Antibiotics (Basel, Switzerland)

    2021  Volume 10, Issue 4

    Abstract: Multidrug ... ...

    Abstract Multidrug resistant
    Language English
    Publishing date 2021-04-15
    Publishing country Switzerland
    Document type Journal Article
    ZDB-ID 2681345-2
    ISSN 2079-6382
    ISSN 2079-6382
    DOI 10.3390/antibiotics10040442
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  4. Article ; Online: Fowlpoxvirus recombinants coding for the CIITA gene increase the expression of endogenous MHC-II and Fowlpox Gag/Pro and Env SIV transgenes.

    Bissa, Massimiliano / Forlani, Greta / Zanotto, Carlo / Tosi, Giovanna / De Giuli Morghen, Carlo / Accolla, Roberto S / Radaelli, Antonia

    PloS one

    2018  Volume 13, Issue 1, Page(s) e0190869

    Abstract: A complete eradication of an HIV infection has never been achieved by vaccination and the search for new immunogens that can induce long-lasting protective responses is ongoing. Avipoxvirus recombinants are host-restricted for replication to avian ... ...

    Abstract A complete eradication of an HIV infection has never been achieved by vaccination and the search for new immunogens that can induce long-lasting protective responses is ongoing. Avipoxvirus recombinants are host-restricted for replication to avian species and they do not have the undesired side effects induced by vaccinia recombinants. In particular, Fowlpox (FP) recombinants can express transgenes over long periods and can induce protective immunity in mammals, mainly due to CD4-dependent CD8+ T cells. In this context, the class II transactivator (CIITA) has a pivotal role in triggering the adaptive immune response through induction of the expression of class-II major histocompatibility complex molecule (MHC-II), that can present antigens to CD4+ T helper cells. Here, we report on construction of novel FPgp and FPenv recombinants that express the highly immunogenic SIV Gag-pro and Env structural antigens. Several FP-based recombinants, with single or dual genes, were also developed that express CIITA, driven from H6 or SP promoters. These recombinants were used to infect CEF and Vero cells in vitro and determine transgene expression, which was evaluated by real-time PCR and Western blotting. Subcellular localisation of the different proteins was evaluated by confocal microscopy, whereas HLA-DR or MHC-II expression was measured by flow cytometry. Fowlpox recombinants were also used to infect syngeneic T/SA tumour cells, then injected into Balb/c mice to elicit MHC-II immune response and define the presentation of the SIV transgene products in the presence or absence of FPCIITA. Antibodies to Env were measured by ELISA. Our data show that the H6 promoter was more efficient than SP to drive CIITA expression and that CIITA can enhance the levels of the gag/pro and env gene products only when infection is performed by FP single recombinants. Also, CIITA expression is higher when carried by FP single recombinants than when combined with FPgp or FPenv constructs and can induce HLA-DR cell surface expression. However, in-vivo experiments did not show any significant increase in the humoral response. As CIITA already proved to elicit immunogenicity by improving antigen presentation, further in-vivo experiments should be performed to increase the immune responses. The use of prime/boost immunisation protocols and the oral administration route of the recombinants may enhance the immunogenicity of Env peptides presented by MHC-II and provide CD4+ T-cell stimulation.
    MeSH term(s) AIDS Vaccines/genetics ; AIDS Vaccines/immunology ; Animals ; Blotting, Western ; CD4-Positive T-Lymphocytes/immunology ; CD8-Positive T-Lymphocytes/immunology ; Cell Line ; Chick Embryo ; Enzyme-Linked Immunosorbent Assay ; Genes, Viral ; HIV Infections/immunology ; HIV Infections/prevention & control ; Humans ; Major Histocompatibility Complex/genetics ; Mice ; Mice, Inbred BALB C ; Microscopy, Confocal ; Nuclear Proteins/genetics ; Poxviridae/genetics ; Promoter Regions, Genetic ; Real-Time Polymerase Chain Reaction ; Recombination, Genetic ; Simian Immunodeficiency Virus/genetics ; Trans-Activators/genetics ; Transgenes
    Chemical Substances AIDS Vaccines ; MHC class II transactivator protein ; Nuclear Proteins ; Trans-Activators
    Language English
    Publishing date 2018
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ISSN 1932-6203
    ISSN (online) 1932-6203
    DOI 10.1371/journal.pone.0190869
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  5. Article: Benzamide Derivatives Targeting the Cell Division Protein FtsZ: Modifications of the Linker and the Benzodioxane Scaffold and Their Effects on Antimicrobial Activity.

    Straniero, Valentina / Suigo, Lorenzo / Casiraghi, Andrea / Sebastián-Pérez, Victor / Hrast, Martina / Zanotto, Carlo / Zdovc, Irena / De Giuli Morghen, Carlo / Radaelli, Antonia / Valoti, Ermanno

    Antibiotics (Basel, Switzerland)

    2020  Volume 9, Issue 4

    Abstract: Filamentous temperature-sensitive Z (FtsZ) is a prokaryotic protein with an essential role in the bacterial cell division process. It is widely conserved and expressed in both Gram-positive and Gram-negative strains. In the last decade, several research ... ...

    Abstract Filamentous temperature-sensitive Z (FtsZ) is a prokaryotic protein with an essential role in the bacterial cell division process. It is widely conserved and expressed in both Gram-positive and Gram-negative strains. In the last decade, several research groups have pointed out molecules able to target FtsZ in
    Language English
    Publishing date 2020-04-04
    Publishing country Switzerland
    Document type Journal Article
    ZDB-ID 2681345-2
    ISSN 2079-6382
    ISSN 2079-6382
    DOI 10.3390/antibiotics9040160
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  6. Article ; Online: Fowlpox virus recombinants expressing HPV-16 E6 and E7 oncogenes for the therapy of cervical carcinoma elicit humoral and cell-mediated responses in rabbits

    Pacchioni Sole / Pozzi Eleana / Radaelli Antonia / Zanotto Carlo / Morghen Carlo

    Journal of Translational Medicine, Vol 8, Iss 1, p

    2010  Volume 40

    Abstract: Abstract Background Around half million new cases of cervical cancer arise each year, making the development of an effective therapeutic vaccine against HPV a high priority. As the E6 and E7 oncoproteins are expressed in all HPV-16 tumour cells, vaccines ...

    Abstract Abstract Background Around half million new cases of cervical cancer arise each year, making the development of an effective therapeutic vaccine against HPV a high priority. As the E6 and E7 oncoproteins are expressed in all HPV-16 tumour cells, vaccines expressing these proteins might clear an already established tumour and support the treatment of HPV-related precancerous lesions. Methods Three different immunisation regimens were tested in a pre-clinical trial in rabbits to evaluate the humoral and cell-mediated responses of a putative HPV-16 vaccine. Fowlpoxvirus (FP) recombinants separately expressing the HPV-16 E6 (FP E6 ) and E7 (FP E7 ) transgenes were used for priming, followed by E7 protein boosting. Results All of the protocols were effective in eliciting a high antibody response. This was also confirmed by interleukin-4 production, which increased after simultaneous priming with both FP E6 and FP E7 and after E7 protein boost. A cell-mediated immune response was also detected in most of the animals. Conclusion These results establish a preliminary profile for the therapy with the combined use of avipox recombinants, which may represent safer immunogens than vaccinia-based vectors in immuno-compromised individuals, as they express the transgenes in most mammalian cells in the absence of a productive replication.
    Keywords Medicine ; R
    Subject code 616 ; 610
    Language English
    Publishing date 2010-04-01T00:00:00Z
    Publisher BMC
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  7. Article ; Online: 2,6-Difluorobenzamide Inhibitors of Bacterial Cell Division Protein FtsZ: Design, Synthesis, and Structure-Activity Relationships.

    Straniero, Valentina / Zanotto, Carlo / Straniero, Letizia / Casiraghi, Andrea / Duga, Stefano / Radaelli, Antonia / De Giuli Morghen, Carlo / Valoti, Ermanno

    ChemMedChem

    2017  Volume 12, Issue 16, Page(s) 1303–1318

    Abstract: A wide variety of drug-resistant microorganisms are continuously emerging, restricting the therapeutic options for common bacterial infections. Antimicrobial agents that were originally potent are now no longer helpful, due to their weak or null activity ...

    Abstract A wide variety of drug-resistant microorganisms are continuously emerging, restricting the therapeutic options for common bacterial infections. Antimicrobial agents that were originally potent are now no longer helpful, due to their weak or null activity toward these antibiotic-resistant bacteria. In addition, none of the recently approved antibiotics affect innovative targets, resulting in a need for novel drugs with innovative antibacterial mechanisms of action. The essential cell division protein filamentous temperature-sensitive Z (FtsZ) has emerged as a possible target, thanks to its ubiquitous expression and its homology to eukaryotic β-tubulin. In the latest years, several compounds were shown to interact with this prokaryotic protein and selectively inhibit bacterial cell division. Recently, our research group developed interesting derivatives displaying good antibacterial activities against methicillin-resistant Staphylococcus aureus, as well as vancomycin-resistant Enterococcus faecalis and Mycobacterium tuberculosis. The aim of the present study was to summarize the structure-activity relationships of differently substituted heterocycles, linked by a methylenoxy bridge to the 2,6-difluorobenzamide, and to validate FtsZ as the real target of this class of antimicrobials.
    MeSH term(s) Anti-Bacterial Agents/chemical synthesis ; Anti-Bacterial Agents/chemistry ; Anti-Bacterial Agents/pharmacology ; Bacteria/drug effects ; Bacteria/metabolism ; Bacterial Proteins/antagonists & inhibitors ; Bacterial Proteins/metabolism ; Benzamides/chemical synthesis ; Benzamides/chemistry ; Benzamides/pharmacology ; Cell Line ; Cell Survival/drug effects ; Cytoskeletal Proteins/antagonists & inhibitors ; Cytoskeletal Proteins/metabolism ; Drug Design ; Enterococcus faecalis/drug effects ; Enterococcus faecalis/metabolism ; Humans ; Methicillin-Resistant Staphylococcus aureus/drug effects ; Methicillin-Resistant Staphylococcus aureus/metabolism ; Microbial Sensitivity Tests ; Mycobacterium tuberculosis/drug effects ; Mycobacterium tuberculosis/metabolism ; Structure-Activity Relationship
    Chemical Substances Anti-Bacterial Agents ; Bacterial Proteins ; Benzamides ; Cytoskeletal Proteins ; FtsZ protein, Bacteria ; 2,6-difluorobenzamide (SJ2RN214MK)
    Language English
    Publishing date 2017-08-22
    Publishing country Germany
    Document type Journal Article
    ZDB-ID 2218496-X
    ISSN 1860-7187 ; 1860-7179
    ISSN (online) 1860-7187
    ISSN 1860-7179
    DOI 10.1002/cmdc.201700201
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  8. Article ; Online: Benzodioxane-Benzamides as Antibacterial Agents: Computational and SAR Studies to Evaluate the Influence of the 7-Substitution in FtsZ Interaction.

    Straniero, Valentina / Sebastián-Pérez, Victor / Hrast, Martina / Zanotto, Carlo / Casiraghi, Andrea / Suigo, Lorenzo / Zdovc, Irena / Radaelli, Antonia / De Giuli Morghen, Carlo / Valoti, Ermanno

    ChemMedChem

    2019  Volume 15, Issue 2, Page(s) 195–209

    Abstract: FtsZ is a crucial prokaryotic protein involved in bacterial cell replication. It recently arose as a promising target in the search for antimicrobial agents able to fight antimicrobial resistance. In this work, going on with our structure-activity ... ...

    Abstract FtsZ is a crucial prokaryotic protein involved in bacterial cell replication. It recently arose as a promising target in the search for antimicrobial agents able to fight antimicrobial resistance. In this work, going on with our structure-activity relationship (SAR) study, we developed variously 7-substituted 1,4-benzodioxane compounds, linked to the 2,6-difluorobenzamide by a methylenoxy bridge. Compounds exhibit promising antibacterial activities not only against multidrug-resistant Staphylococcus aureus, but also on mutated Escherichia coli strains, thus enlarging their spectrum of action toward Gram-negative bacteria as well. Computational studies elucidated, through a validated FtsZ binding protocol, the structural features of new promising derivatives as FtsZ inhibitors.
    MeSH term(s) Anti-Bacterial Agents/chemical synthesis ; Anti-Bacterial Agents/chemistry ; Anti-Bacterial Agents/pharmacology ; Bacterial Proteins/antagonists & inhibitors ; Bacterial Proteins/metabolism ; Benzamides/chemistry ; Benzamides/pharmacology ; Benzodioxoles/chemistry ; Benzodioxoles/pharmacology ; Cytoskeletal Proteins/antagonists & inhibitors ; Cytoskeletal Proteins/metabolism ; Dose-Response Relationship, Drug ; Drug Resistance, Multiple, Bacterial/drug effects ; Escherichia coli/drug effects ; Microbial Sensitivity Tests ; Molecular Docking Simulation ; Molecular Structure ; Staphylococcus aureus/drug effects ; Structure-Activity Relationship
    Chemical Substances Anti-Bacterial Agents ; Bacterial Proteins ; Benzamides ; Benzodioxoles ; Cytoskeletal Proteins ; FtsZ protein, Bacteria
    Language English
    Publishing date 2019-12-11
    Publishing country Germany
    Document type Journal Article
    ZDB-ID 2218496-X
    ISSN 1860-7187 ; 1860-7179
    ISSN (online) 1860-7187
    ISSN 1860-7179
    DOI 10.1002/cmdc.201900537
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  9. Article ; Online: Construction and characterisation of a recombinant fowlpox virus that expresses the human papilloma virus L1 protein

    Zanotto Carlo / Pozzi Eleana / Pacchioni Sole / Bissa Massimiliano / De Giuli Morghen Carlo / Radaelli Antonia

    Journal of Translational Medicine, Vol 9, Iss 1, p

    2011  Volume 190

    Abstract: Abstract Background Human papilloma virus (HPV)-16 is the most prevalent high-risk mucosal genotype. Virus-like-particle (VLP)-based immunogens developed recently have proven to be successful as prophylactic HPV vaccines, but are still too expensive for ... ...

    Abstract Abstract Background Human papilloma virus (HPV)-16 is the most prevalent high-risk mucosal genotype. Virus-like-particle (VLP)-based immunogens developed recently have proven to be successful as prophylactic HPV vaccines, but are still too expensive for developing countries. Although vaccinia viruses expressing the HPV-16 L1 protein (HPV-L1) have been studied, fowlpox-based recombinants represent efficient and safer vectors for immunocompromised hosts due to their ability to elicit a complete immune response and their natural host-range restriction to avian species. Methods A new fowlpox virus recombinant encoding HPV-L1 (FP L1 ) was engineered and evaluated for the correct expression of HPV-L1 in vitro , using RT-PCR, immunoprecipitation, Western blotting, electron microscopy, immunofluorescence, and real-time PCR assays. Results The FP L1 recombinant correctly expresses HPV-L1 in mammalian cells, which are non-permissive for the replication of this vector. Conclusion This FP L1 recombinant represents an appropriate immunogen for expression of HPV-L1 in human cells. The final aim is to develop a safe, immunogenic, and less expensive prophylactic vaccine against HPV.
    Keywords Medicine ; R
    Subject code 570
    Language English
    Publishing date 2011-11-01T00:00:00Z
    Publisher BMC
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  10. Article ; Online: Prime-boost therapeutic vaccination in mice with DNA/DNA or DNA/Fowlpox virus recombinants expressing the Human Papilloma Virus type 16 E6 and E7 mutated proteins fused to the coat protein of Potato virus X.

    Illiano, Elena / Bissa, Massimiliano / Paolini, Francesca / Zanotto, Carlo / De Giuli Morghen, Carlo / Franconi, Rosella / Radaelli, Antonia / Venuti, Aldo

    Virus research

    2016  Volume 225, Page(s) 82–90

    Abstract: The therapeutic antitumor potency of a prime-boost vaccination strategy was explored, based on the mutated, nontransforming forms of the E6 ( ... ...

    Abstract The therapeutic antitumor potency of a prime-boost vaccination strategy was explored, based on the mutated, nontransforming forms of the E6 (E6
    Language English
    Publishing date 2016-10-02
    Publishing country Netherlands
    Document type Journal Article
    ZDB-ID 605780-9
    ISSN 1872-7492 ; 0168-1702
    ISSN (online) 1872-7492
    ISSN 0168-1702
    DOI 10.1016/j.virusres.2016.09.011
    Database MEDical Literature Analysis and Retrieval System OnLINE

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