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Microorganisms
Special Issues
Phage Display—Perspectives and Translational Applications
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Phage Display—Perspectives and Translational Applications

A special issue of Microorganisms (ISSN 2076-2607).

Deadline for manuscript submissions: closed (31 December 2020) | Viewed by 17874

Special Issue Editor

Dr. Piergiuseppe De Berardinis

E-Mail Website
Guest Editor
Institute of Biochemistry and Cell Biology, CNR, Naples, Italy
Interests: immunological studies for the formulation of new immunotherapeutic strategies and innovative research products; T lymphocytes; immune response; phage display

Special Issue Information

Dear Colleagues,

Thirty-four years after the first description of phage display technology, George Smith, its inventor, has been awarded the Nobel prize in Chemistry. In these intervening years, we have observed an explosion in the application of this technology, which is generating renewed interests for translational usage. Filamentous bacteriophages combine a particulate nature with adjuvant properties and thus may represent an ideal nanoparticle for medical applications. In recent years, it has been proposed for drug targeting and has attracted great interest for its potential immunotherapeutic formulations. However, much research is needed before bacteriophage can be realized as a device ready off the shelf for human therapy or prophylaxis and such usage faces substantial challenges. Most of the research has been performed so far in academic laboratories, with little emphasis on patenting. Moreover, a major obstacle to the therapeutic use of bacteriophages is their characteristic of being “living” organisms, able to interact with the endogenous microbiome, with unpredictable outcomes at this stage of knowledge.

The aim of this Special Issue is to update recent knowledge on the use of filamentous bacteriophages and provide a platform for scientists working in the field.

This proposed issue is open to receive manuscripts on the following topics:

(1) Basic insights: phage display vehicles, expression, screening, and strategies for affinity evolution

(2) Practicalities, patenting, and safety issues

(3) Use for drug targeting, diagnostic strategies, and immunotherapeutic formulations

Dr. Piergiuseppe De Berardinis
Guest Editor

Manuscript Submission Information

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. Microorganisms is an international peer-reviewed open access monthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 2700 CHF (Swiss Francs). Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

  • Filamentous bacteriophage
  • Phage display
  • Drug delivery
  • Cell targeting
  • Nanotechnology
  • Inoviridae
  • Immunotherapeutic strategies
  • Bacteriophage adjuvanticity

Published Papers (5 papers)

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15 pages, 2421 KiB  
Article
Phage Display Detection of Mimotopes that Are Shared Epitopes of Clinically and Epidemiologically Relevant Enterobacteria
by Armando Navarro, Delia Licona-Moreno, Alejandro Monsalvo-Reyes, Ulises Hernández-Chiñas and Carlos A. Eslava-Campos
Microorganisms 2020, 8(5), 780; https://doi.org/10.3390/microorganisms8050780 - 22 May 2020
Cited by 3 | Viewed by 2800
Abstract
Background: Escherichia coli and Salmonella are etiologic agents of intestinal infections. A previous study showed the presence of shared epitopes between lipopolysaccharides (LPSs) of E. coli O157 and Salmonella. Aim: Using phage display, the aim of this study is to identify mimotopes [...] Read more.
Background: Escherichia coli and Salmonella are etiologic agents of intestinal infections. A previous study showed the presence of shared epitopes between lipopolysaccharides (LPSs) of E. coli O157 and Salmonella. Aim: Using phage display, the aim of this study is to identify mimotopes of shared epitopes in different enterobacterial LPSs. Methods: We use anti-LPS IgG from E. coli O157 and Salmonella to select peptide mimotopes of the M13 phage. The amino acid sequence of the mimotopes is used to synthesize peptides, which are in turn used to immunize rabbits. The antibody response of the resulting sera against the LPSs and synthetic peptides (SPs) is analyzed by ELISA and by Western blot assays, indicating that LPS sites are recognized by the same antibody. In a complementary test, the reactions of human serum samples obtained from the general population against the SPs and LPSs are also analyzed. Results: From the last biopanning phase, sixty phagotopes are selected. The analysis of the peptide mimotope amino acid sequences shows that in 4 of them the S/N/A/PF motif is a common sequence. Antibodies from the sera of immunized rabbits with SP287/3, SP459/1, SP308/3, and SP073/14 react against both their own peptide and the different LPSs. The Western blot test shows a sera reaction against both the lateral chains and the cores of the LPSs. The analysis of the human sera shows a response against the SPs and LPSs. Conclusion: The designed synthetic peptides are mimotopes of LPS epitopes of Salmonella and E. coli that possess immunogenic capacity. These mimotopes could be considered for use in the design of vaccines against both enterobacteria. Full article
(This article belongs to the Special Issue Phage Display—Perspectives and Translational Applications)
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14 pages, 3361 KiB  
Article
Recombinant Filamentous Bacteriophages Encapsulated in Biodegradable Polymeric Microparticles for Stimulation of Innate and Adaptive Immune Responses
by Rezvan Jamaledin, Rossella Sartorius, Concetta Di Natale, Raffaele Vecchione, Piergiuseppe De Berardinis and Paolo Antonio Netti
Microorganisms 2020, 8(5), 650; https://doi.org/10.3390/microorganisms8050650 - 29 Apr 2020
Cited by 30 | Viewed by 3828
Abstract
Escherichia coli filamentous bacteriophages (M13, f1, or fd) have attracted tremendous attention from vaccinologists as a promising immunogenic carrier and vaccine delivery vehicle with vast possible applications in the development of vaccines. The use of fd bacteriophage as an antigen delivery system is [...] Read more.
Escherichia coli filamentous bacteriophages (M13, f1, or fd) have attracted tremendous attention from vaccinologists as a promising immunogenic carrier and vaccine delivery vehicle with vast possible applications in the development of vaccines. The use of fd bacteriophage as an antigen delivery system is based on a modification of bacteriophage display technology. In particular, it is designed to express multiple copies of exogenous peptides (or polypeptides) covalently linked to viral capsid proteins. This study for the first time proposes the use of microparticles (MPs) made of poly (lactic-co-glycolic acid) (PLGA) to encapsulate fd bacteriophage. Bacteriophage–PLGA MPs were synthesized by a water in oil in water (w1/o/w2) emulsion technique, and their morphological properties were analyzed by confocal and scanning electron microscopy (SEM). Moreover, phage integrity, encapsulation efficiency, and release were investigated. Using recombinant bacteriophages expressing the ovalbumin (OVA) antigenic determinant, we demonstrated the immunogenicity of the encapsulated bacteriophage after being released by MPs. Our results reveal that encapsulated bacteriophages are stable and retain their immunogenic properties. Bacteriophage-encapsulated PLGA microparticles may thus represent an important tool for the development of different bacteriophage-based vaccine platforms. Full article
(This article belongs to the Special Issue Phage Display—Perspectives and Translational Applications)
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12 pages, 2528 KiB  
Communication
Analysis of the Consolidation Phase of Immunological Memory within the IgG Response to a B Cell Epitope Displayed on a Filamentous Bacteriophage
by Francesca Mantile, Angelo Capasso, Piergiuseppe De Berardinis and Antonella Prisco
Microorganisms 2020, 8(4), 564; https://doi.org/10.3390/microorganisms8040564 - 14 Apr 2020
Cited by 2 | Viewed by 1771
Abstract
Immunological memory can be defined as the ability to mount a response of greater magnitude and with faster kinetics upon re-encounter of the same antigen. We have previously reported that a booster dose of a protein antigen given 15 days after the first [...] Read more.
Immunological memory can be defined as the ability to mount a response of greater magnitude and with faster kinetics upon re-encounter of the same antigen. We have previously reported that a booster dose of a protein antigen given 15 days after the first dose interferes with the development of memory, i.e., with the ability to mount an epitope-specific IgG response of greater magnitude upon re-encounter of the same antigen. We named the time-window during which memory is vulnerable to disruption a “consolidation phase in immunological memory”, by analogy with the memory consolidation processes that occur in the nervous system to stabilize memory traces. In this study, we set out to establish if a similar memory consolidation phase occurs in the IgG response to a B cell epitope displayed on a filamentous bacteriophage. To this end, we have analyzed the time-course of anti-β-amyloid IgG titers in mice immunized with prototype Alzheimer’s Disease vaccine fdAD(2-6), which consists of a fd phage that displays the B epitope AEFRH of β -amyloid at the N-terminus of the Major Capsid Protein. A booster dose of phage fdAD(2-6) given 15 days after priming significantly reduced the ratio between the magnitude of the secondary and primary IgG response to β-amyloid. This analysis confirms, in a phage vaccine, a consolidation phase in immunological memory, occurring two weeks after priming. Full article
(This article belongs to the Special Issue Phage Display—Perspectives and Translational Applications)
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18 pages, 2775 KiB  
Article
Application of a Broad Range Lytic Phage LPST94 for Biological Control of Salmonella in Foods
by Md. Sharifull Islam, Yang Zhou, Lu Liang, Ishatur Nime, Ting Yan, Stephan P. Willias, Md. Zakaria Mia, Weicheng Bei, Ian F. Connerton, Vincent A. Fischetti and Jinquan Li
Microorganisms 2020, 8(2), 247; https://doi.org/10.3390/microorganisms8020247 - 13 Feb 2020
Cited by 36 | Viewed by 4959
Abstract
Salmonella, one of the most common food-borne pathogens, is a significant public health and economic burden worldwide. Lytic phages are viable alternatives to conventional technologies for pathogen biocontrol in food products. In this study, 40 Salmonella phages were isolated from environmentally sourced [...] Read more.
Salmonella, one of the most common food-borne pathogens, is a significant public health and economic burden worldwide. Lytic phages are viable alternatives to conventional technologies for pathogen biocontrol in food products. In this study, 40 Salmonella phages were isolated from environmentally sourced water samples. We characterized the lytic range against Salmonella and among all isolates, phage LPST94 showed the broadest lytic spectrum and the highest lytic activity. Electron microscopy and genome sequencing indicated that LPST94 belongs to the Ackermannviridae family. Further studies showed this phage is robust, tolerating a wide range of pH (4–12) and temperature (30–60 °C) over 60 min. The efficacy of phage LPST94 as a biological control agent was evaluated in various food products (milk, apple juice, chicken breast, and lettuce) inoculated with non-typhoidal Salmonella species at different temperatures. Interestingly, the anti-Salmonella efficacy of phage LPST94 was greater at 4 °C than 25 °C, although the efficacy varied between different food models. Adding phage LPST94 to Salmonella inoculated milk decreased the Salmonella count by 3 log10 CFU/mL at 4 °C and 0.84 to 2.56 log10 CFU/mL at 25 °C using an MOI of 1000 and 10000, respectively. In apple juice, chicken breast, and lettuce, the Salmonella count was decreased by 3 log10 CFU/mL at both 4 °C and 25 °C after applying phage LPST94 at an MOI of 1000 and 10,000, within a timescale of 48 h. The findings demonstrated that phage LPST94 is a promising candidate for biological control agents against pathogenic Salmonella and has the potential to be applied across different food matrices. Full article
(This article belongs to the Special Issue Phage Display—Perspectives and Translational Applications)
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10 pages, 1547 KiB  
Brief Report
Bacteriophages Promote Metabolic Changes in Bacteria Biofilm
by Marina Papaianni, Paola Cuomo, Andrea Fulgione, Donatella Albanese, Monica Gallo, Debora Paris, Andrea Motta, Domenico Iannelli and Rosanna Capparelli
Microorganisms 2020, 8(4), 480; https://doi.org/10.3390/microorganisms8040480 - 28 Mar 2020
Cited by 12 | Viewed by 3701
Abstract
Bacterial biofilm provides bacteria with resistance and protection against conventional antimicrobial agents and the host immune system. Bacteriophages are known to move across the biofilm to make it permeable to antimicrobials. Mineral hydroxyapatite (HA) can improve the lytic activity of bacteriophages, and, together [...] Read more.
Bacterial biofilm provides bacteria with resistance and protection against conventional antimicrobial agents and the host immune system. Bacteriophages are known to move across the biofilm to make it permeable to antimicrobials. Mineral hydroxyapatite (HA) can improve the lytic activity of bacteriophages, and, together with eicosanoic acid (C20:0), can destroy the biofilm structure. Here, we demonstrate the efficacy of the combined use of phage, HA and C20:0 against Xanthomonas campestris pv campestris (Xcc) biofilm. We used nuclear magnetic resonance (NMR)-based metabolomics to investigate the molecular determinants related to the lytic action, aiming at identifying the metabolic pathways dysregulated by phage treatment. Furthermore, we identified specific markers (amino acids, lactate and galactomannan) which are involved in the control of biofilm stability. Our data show that Xccφ1, alone or in combination with HA and C20:0, interferes with the metabolic pathways involved in biofilm formation. The approach described here might be extended to other biofilm-producing bacteria. Full article
(This article belongs to the Special Issue Phage Display—Perspectives and Translational Applications)
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