Functional Pentameric Formation via Coexpression of the Escherichia coli Heat-Labile Enterotoxin B Subunit and Its Fusion Protein Subunit with a Neutralizing Epitope of ApxIIA Exotoxin Improves the Mucosal Immunogenicity and Protection against Challenge by Actinobacillus pleuropneumoniae

Kim, Jung-Mi; Park, Seung-Moon; Kim, Jung-Ae; Park, Jin‐Ah; Yi, Min‐Hee; Kim, Nan-Sun; Bae, Jong-Lye; Park, Sung Goo; Jang, Yong-Suk; Yang, Moon-Sik; Kim, Dae-Hyuk

doi:10.1128/cvi.05230-11

Cited by 17 publications

(17 citation statements)

References 64 publications

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“…LTB alone is not toxic but able to induce a potent mucosal immunogenic activity. In addition, LTB can exert a mucosal adjuvant activity for coadministered unrelated antigens [ 24 ]. Although the mechanism of the adjuvant effects of LTB is not known, such activity strictly depends on LTB ability to bind to its cell receptor GM1, and therefore on the pentameric association of five B subunits [ 24 ].…”

Section: Resultsmentioning

confidence: 99%

“…In addition, LTB can exert a mucosal adjuvant activity for coadministered unrelated antigens [ 24 ]. Although the mechanism of the adjuvant effects of LTB is not known, such activity strictly depends on LTB ability to bind to its cell receptor GM1, and therefore on the pentameric association of five B subunits [ 24 ]. When LTB is expressed in a heterologous bacterium fused to an anchor protein, pentamer formation is impaired [ 25 ].…”

Section: Resultsmentioning

confidence: 99%

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Non-recombinant display of the B subunit of the heat labile toxin of Escherichia coli on wild type and mutant spores of Bacillus subtilis

et al. 2013

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BackgroundMucosal infections are a major global health problem and it is generally accepted that mucosal vaccination strategies, able to block infection at their entry site, would be preferable with respect to other prevention approaches. However, there are still relatively few mucosal vaccines available, mainly because of the lack of efficient delivery systems and of mucosal adjuvants. Recombinant bacterial spores displaying a heterologous antigen have been shown to induce protective immune responses and, therefore, proposed as a mucosal delivery system. A non-recombinant approach has been recently developed and tested to display antigens and enzymes.ResultsWe report that the binding subunit of the heat-labile toxin (LTB) of Escherichia coli efficiently adsorbed on the surface of Bacillus subtilis spores. When nasally administered to groups of mice, spore-adsorbed LTB was able to induce a specific immune response with the production of serum IgG, fecal sIgA and of IFN-γ in spleen and mesenteric lymph nodes (MLN) of the immunized animals. Dot blotting experiments showed that the non-recombinant approach was more efficient than the recombinant system in displaying LTB and that the efficiency of display could be further increased by using mutant spores with an altered surface. In addition, immunofluorescence microscopy experiments showed that only when displayed on the spore surface by the non-recombinant approach LTB was found in its native, pentameric form.ConclusionOur results indicate that non-recombinant spores displaying LTB pentamers can be administered by the nasal route to induce a Th1-biased, specific immune response. Mutant spores with an altered coat are more efficient than wild type spores in adsorbing the antigen, allowing the use of a reduced number of spores in immunization procedures. Efficiency of display, ability to display the native form of the antigen and to induce a specific immune response propose this non-recombinant delivery system as a powerful mucosal vaccine delivery approach.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Non-recombinant display of the B subunit of the heat labile toxin of Escherichia coli on wild type and mutant spores of Bacillus subtilis

et al. 2013

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“…pleuropneumoniae is the etiologic agent of porcine pleuropneumonia in hyperacute, acute or chronic form [36]- [40]. Positive results based on the apxIV gene amplification indicate the presence of this bacterium in almost all samples, with the exception of urine samples.…”

Section: Discussionmentioning

confidence: 99%

Porcine Respiratory Pathogens in Swine Farms Environment in Mexico

Loera-Muro¹,

Loera‐Muro²,

Morfín-Mata³

et al. 2014

OJAS

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Respiratory pathogens are the main health problem in the swine industry worldwide. These pathogens are transmitted by direct contact between animals or by aerosols and however are not well known yet, if the environment works as its reservoir, inoculum and/or dispersion medium. The objective of this study was to determine the presence of respiratory pathogens in environmental samples from swine farms in Aguascalientes, Mexico, through of PCR and RT-PCR techniques. The bacteria Actinobacillus pleuropneumoniae and Pasteurella multocida were found viable in samples from water, food, soil and air. Streptococcus suis was found in a viable state in water samples. Haemophilus parasuis, Porcine Reproductive and Respiratory Syndrome virus and Swine Influenza virus (H1N1 and H3N2) were detected in drinking water samples. Mycoplasma hyopneumoniae and Porcine Circovirus type 2 (PCV2) were not detected in environmental samples. These results suggest that the environment of the farms acts as a reservoir, inoculum and/or vehicle of dispersion for these pathogens except for M. hyopneumoniae and PCV2.

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“…Application of the toxin family to protein delivery has been mostly limited to vaccine development, e.g. 24 , and, very rarely, non-vaccine protein cargo 25 – 27 . The limitation is principally due to a lack of detailed understanding of the requirements for co-assembly with generic cargo.…”

Section: Introductionmentioning

confidence: 99%

Engineering an AB5 Protein Carrier

Lichtenstein

Höcker

2018

Sci Rep

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The promise of biologic therapeutics is hindered by the challenge to deliver their activity to biochemically relevant sites within diseased cells. The favourable application of the natural protein carriers of the AB5 toxin family to this challenge has been restricted owing to still unresolved requirements for assembling non-native cargo into carrier complexes. Here, we clarify the properties of fusion peptides which allow co-assembly of a selected fluorescent protein cargo with the non-toxic B subunit of a heat-labile enterotoxin. We establish the influence of sequence length, sequence identity and secondary structure of these linking domains on the assembly and disassembly of the complexes. Through our engineering framework we identify several non-native, reduced length fusion sequences that robustly assemble with the native carriers, maintain their ability to deliver protein cargo to cells, and demonstrate substantially refined in vitro properties. Constructs based upon these sequences should prove directly applicable to a variety of protein delivery challenges, and the described design framework should find immediate application to other members of the AB5 protein carrier family.

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Functional Pentameric Formation via Coexpression of the Escherichia coli Heat-Labile Enterotoxin B Subunit and Its Fusion Protein Subunit with a Neutralizing Epitope of ApxIIA Exotoxin Improves the Mucosal Immunogenicity and Protection against Challenge by Actinobacillus pleuropneumoniae

Cited by 17 publications

References 64 publications

Non-recombinant display of the B subunit of the heat labile toxin of Escherichia coli on wild type and mutant spores of Bacillus subtilis

Non-recombinant display of the B subunit of the heat labile toxin of Escherichia coli on wild type and mutant spores of Bacillus subtilis

Porcine Respiratory Pathogens in Swine Farms Environment in Mexico

Engineering an AB5 Protein Carrier

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