The amino acid selected for generating mutant TbpB antigens defective in binding transferrin can compromise the in vivo protective capacity

Guizzo, João Antônio; Chaudhuri, Somshukla; Prigol, Simone Ramos; Yu, Rong-hua; Dazzi, C; Balbinott, Natalia; Frandoloso, Gabriela Paraboni; Kreutz, Luiz Carlos; Frandoloso, Rafael; Schryvers, Anthony B.

doi:10.1038/s41598-018-25685-1

Cited by 21 publications

(35 citation statements)

References 44 publications

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“…The demonstration that the diversity of TbpBs was distributed into three phylogenetic clusters that were not strongly associated with the time of isolation, geographical region or even species (present in G. parasuis, Actinobacillus pleuropneumoniae and A. suis) (12) suggested that it would be possible to design a TbpB-based vaccine effective against these three porcine pathogens. The TbpB Y167A antigen was able to provide protection against strains expressing a homologous or heterologous TbpB from the same phylogenetic cluster (13) (11). These results suggest that it will be possible to generate complete protection against Glässer's disease regardless of the capsular type of this bacterium and extend the protective effect to Actinobacillus pleuropneumoniae and A. suis due to the shared distribution of variants amongst these species (12).…”

Section: Introductionmentioning

confidence: 84%

“…Growth of G. parasuis was as described previously (13). An aliquot of SV7 reference strain 174, which was previously passaged in pigs, was inoculated into supplemented PPLO broth [60 mg/ml nicotinamide adenine dinucleotide (b-NAD, Sigma-Adrich, USA) and 2.5 mg/ml D-glucose (Sigma-Aldrich, USA)] and grown with shaking (250 rpm/37°C) to achieve an optical density of 0.5 at 600 nm.…”

Section: G Parasuis Strain and Growth Conditionsmentioning

confidence: 99%

“…The gene encoding the Y167A variant of the G. parasuis SV7 TbpB was cloned into a custom T7 expression vector containing an N-terminal polyhistidine tag and transformed into Escherichia coli strain ER2566 and induced for expression as previously described (13). Briefly, the cultures were grown overnight in autoinduction media, and the protein was produced under lac induction.…”

Section: Antigen Productionmentioning

confidence: 99%

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Proof of Concept for Prevention of Natural Colonization by Oral Needle-Free Administration of a Microparticle Vaccine

Frandoloso

Chaudhuri

Frandoloso³

et al. 2020

Front. Immunol.

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There has been substantial interest in the development of needle-free vaccine administration that has led to a variety of approaches for delivery through the skin for induction of a systemic immune response. The mucosal administration of vaccines has inherently been needle-free, but the simple application of vaccines on the mucosal surface by itself does not lead to mucosal immunity. Since many important bacterial infections develop after initial colonization of the upper respiratory tract of the host, prevention of colonization could not only prevent infection but also eliminate the reservoir of pathogens that reside exclusively in that ecologic niche. This study was designed to provide proof of concept for a needle-free immunization approach that would reduce or eliminate colonization and prevent infection. In order to accomplish this a microparticle vaccine preparation was delivered just below the oral mucosal epithelial cell layer where it would lead to a robust immune response. A vaccine antigen (mutant transferrin binding protein B) shown to be capable of preventing infection in pigs was incorporated into a polyphosphazene microparticle preparation and delivered by a needle-free device to the oral sub-epithelial space of pigs. This vaccination regimen not only provided complete protection from infection after intranasal challenge by Glaesserella parasuis but also eliminated natural colonization by this bacterium. Notably, the complete prevention of natural colonization was dependent upon delivery of the microparticle preparation below the epithelial layer in the oral mucosa as intradermal or intramuscular delivery was not as effective at preventing natural colonization. This study also demonstrated that a primary immunization in the presence of maternal antibody limited the resulting antibody response but a robust antibody response after the second immunization indicated that maternal antibody did not prevent induction of B-cell memory.

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Section: Introductionmentioning

confidence: 84%

Section: G Parasuis Strain and Growth Conditionsmentioning

confidence: 99%

Section: Antigen Productionmentioning

confidence: 99%

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Proof of Concept for Prevention of Natural Colonization by Oral Needle-Free Administration of a Microparticle Vaccine

Frandoloso

Chaudhuri

Frandoloso³

et al. 2020

Front. Immunol.

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“…The limit of detection is indicated by the dashed line. targeting surface receptors involved in acquiring from the host iron-binding protein transferrin (16,41). The rationale for targeting surface proteins involved in iron acquisition also underlies recent attempts to target the BauA siderophore receptor from A. baumannii (42,43).…”

Section: Discussionmentioning

confidence: 99%

Hybrid Antigens Expressing Surface Loops of ZnuD From Acinetobacter baumannii Is Capable of Inducing Protection Against Infection

et al. 2020

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Acinetobacter baumannii is an important human pathogen causing substantial mortality in hospitalized patients for which treatment with antibiotics has become problematic due to growing antibiotic resistance. In an attempt to develop alternative strategies for dealing with these serious infections surface antigens are being considered as targets for vaccines or immunotherapy. The surface receptor proteins required for zinc acquisition in Gram-negative bacterial pathogens have been proposed as vaccine targets due to their crucial role for growth in the human host. In this study we selected the putative ZnuD outer membrane receptor from A. baumannii as a target for vaccine development. Due to challenges in production of an integral outer membrane protein for vaccine production, we adopted a recently described hybrid antigen approach in which surface epitopes from the Neisseria meningitidis TbpA receptor protein were displayed on a derivative of the C-lobe of the surface lipoprotein TbpB, named the loopless C-lobe (LCL). A structural model for ZnuD was generated and four surface loops were selected for hybrid antigen production by computational approaches. Hybrid antigens were designed displaying the four selected loops (2, 5, 7, and 11) individually or together in a single hybrid antigen. The hybrid antigens along with ZnuD and the LCL scaffold were produced in the E. coli cytoplasm either as soluble antigens or as inclusion bodies, that were used to generate soluble antigens upon refolding. Mice were immunized with the hybrid antigens, ZnuD or LCL and then used in an A. baumannii sepsis model to evaluate their ability to protect against infection. As expected, the LCL scaffold did not induce a protective immune response, enabling us to attribute observed protection to the displayed loops. Immunization with the refolded ZnuD protein protected 63% of the mice while immunization with hybrid antigens displaying individual loops achieved between 25 and 50% protection. Notably, the mice immunized with the hybrid antigen displaying the four loops were completely protected from infection.

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“…Twenty snatch-farrowed colostrum-deprived piglets (DB Genética Suína, Brazil) selected from five different sows from a high health status herd were obtained and raised as described previously (12). At day 42, piglets were anesthetized using the following drug combination: 0.3 mg/kg of acepromazine (Syntec do Brasil, Brazil), 0.3 mg/kg of midazolam (Laboratório Teuto Brasileiro, Brazil), and 15 mg/kg of Ketamine (Ceva Santé Animale, Brazil) injected by the intramuscular route.…”

Section: Pig Experimental Infection and Clinical Evaluationmentioning

confidence: 99%

New Pathological Lesions Developed in Pigs by a “Non-virulent” Strain of Glaesserella parasuis

et al. 2020

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Glaesserella parasuis is a Gram-negative bacterium that causes Glässer's disease, a common pathology found in young pigs characterized by polyarthritis, polyserositis, and meningitis. The bacterium has 15 known serovars that have been classified by virulence. Serovars 1, 4, 5, and 12 are considered highly virulent and used in most studies. Serovars 3, 6, 7, 9, and 11 are considered avirulent. Recent reports that serovar 7 is an emerging problem in the pig industry indicate that the association of virulence and serovar may not always be reliable. This led us to infect colostrum-deprived piglets with the reference serovar 7 strain (SV7 strain 174) that had been passaged through pigs and characterize the clinical and pathological signs. We observed that SV7 strain 174 caused clinical signs consistent with Glässer's disease in all infected piglets that succumbed to infection for up to day 5 post-infection. Macroscopic and microscopic lesions were consistent with those found in piglets infected with conventional virulent serovars. In addition, we describe novel microscopic lesions associated with Glässer's disease such as endophthalmitis and thymic depletion. Thus, our findings indicate that SV7 strain 174 causes classical signs of Glässer's disease in colostrum-deprived piglets and some caution should be used in employing vaccine strategies based on association between capsular serovar and virulence.

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The amino acid selected for generating mutant TbpB antigens defective in binding transferrin can compromise the in vivo protective capacity

Cited by 21 publications

References 44 publications

Proof of Concept for Prevention of Natural Colonization by Oral Needle-Free Administration of a Microparticle Vaccine

Proof of Concept for Prevention of Natural Colonization by Oral Needle-Free Administration of a Microparticle Vaccine

Hybrid Antigens Expressing Surface Loops of ZnuD From Acinetobacter baumannii Is Capable of Inducing Protection Against Infection

New Pathological Lesions Developed in Pigs by a “Non-virulent” Strain of Glaesserella parasuis

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