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      Oral vaccination with novel Lactococcus lactis mucosal live vector-secreting Brucella lumazine synthase (BLS) protein induces humoral and cellular immune protection against Brucella abortus

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          Abstract

          This work aimed to provide recombinant Lactococcus lactis as a potential live vector for the manufacture of recombinant Brucella abortus (rBLS-Usp45). The sequences of the genes were collected from the GenBank database. Using Vaxijen and ccSOL, the proteins' immunogenicity and solubility were evaluated. Mice were given oral vaccinations with recombinant L. lactis. Anti-BLS-specific IgG antibodies were measured by ELISA assay. Cytokine reactions were examined using real-time PCR and the ELISA technique. The BLS protein was chosen for immunogenicity based on the vaccinology screening findings since it had maximum solubility and antigenic values ​​of 99% and 0.75, respectively. The BLS gene, digested at 477 bp, was electrophoretically isolated to demonstrate that the recombinant plasmid was successfully produced. Protein-level antigen expression showed that the target group produced the 18 kDa-sized BLS protein, whereas the control group did not express any proteins. In the sera of mice given the L. lactis-pNZ8148-BLS-Usp45 vaccine 14 days after priming, there was a significant level of BLS-specific IgG1, IgG2a ( P < 0.001) compared to the PBS control group. Vaccinated mice showed higher levels of IFN-γ, TNFα, IL-4, and IL-10 in samples obtained on days 14 and 28, after receiving the L. lactis-pNZ8148-BLS-Usp45 and IRBA vaccines ( P < 0.001). The inflammatory reaction caused less severe spleen injuries, alveolar edema, lymphocyte infiltration, and morphological damage in the target group's spleen sections. Based on our findings, an oral or subunit-based vaccine against brucellosis might be developed using L. lactis-pNZ8148-BLS-Usp45 as a novel, promising, and safe alternative to the live attenuated vaccines now available.

          Supplementary Information

          The online version contains supplementary material available at 10.1007/s00203-023-03471-6.

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          Pre-expression of a sulfhydryl oxidase significantly increases the yields of eukaryotic disulfide bond containing proteins expressed in the cytoplasm of E.coli

          Van Dat Nguyen, Feras Hatahet, Kirsi Salo … (2011)
          Background Disulfide bonds are one of the most common post-translational modifications found in proteins. The production of proteins that contain native disulfide bonds is challenging, especially on a large scale. Either the protein needs to be targeted to the endoplasmic reticulum in eukaryotes or to the prokaryotic periplasm. These compartments that are specialised for disulfide bond formation have an active catalyst for their formation, along with catalysts for isomerization to the native state. We have recently shown that it is possible to produce large amounts of prokaryotic disulfide bond containing proteins in the cytoplasm of wild-type bacteria such as E. coli by the introduction of catalysts for both of these processes. Results Here we show that the introduction of Erv1p, a sulfhydryl oxidase and a disulfide isomerase allows the efficient formation of natively folded eukaryotic proteins with multiple disulfide bonds in the cytoplasm of E. coli. The production of disulfide bonded proteins was also aided by the use of an appropriate fusion protein to keep the folding intermediates soluble and by choice of media. By combining the pre-expression of a sulfhydryl oxidase and a disulfide isomerase with these other factors, high level expression of even complex disulfide bonded eukaryotic proteins is possible Conclusions Our results show that the production of eukaryotic proteins with multiple disulfide bonds in the cytoplasm of E. coli is possible. The required exogenous components can be put onto a single plasmid vector allowing facile transfer between different prokaryotic strains. These results open up new avenues for the use of E. coli as a microbial cell factory.
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            Recent advances in Brucella abortus vaccines

            Elaine MS Dorneles, Nammalwar Sriranganathan, Andrey Lage (2015)
            Brucella abortus vaccines play a central role in bovine brucellosis control/eradication programs and have been successfully used worldwide for decades. Strain 19 and RB51 are the approved B. abortus vaccines strains most commonly used to protect cattle against infection and abortion. However, due to some drawbacks shown by these vaccines much effort has been undertaken for the development of new vaccines, safer and more effective, that could also be used in other susceptible species of animals. In this paper, we present a review of the main aspects of the vaccines that have been used in the brucellosis control over the years and the current research advances in the development of new B. abortus vaccines.
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              Vaccines, reverse vaccinology, and bacterial pathogenesis.

              Isabel Delany, Rino Rappuoli, Kate L. Seib (2013)
              Advances in genomics and innovative strategies such as reverse vaccinology have changed the concepts and approaches to vaccine candidate selection and design. Genome mining and blind selection of novel antigens provide a novel route to investigate the mechanisms that underpin pathogenesis. The resulting lists of novel candidates are revealing new aspects of pathogenesis of target organisms, which in turn drives the rational design of optimal vaccine antigens. Here we use the discovery, characterization, and exploitation of fHbp, a vaccine candidate and key virulence factor of meningococcus, as an illustrative case in point. Applying genomic approaches to study both the pathogen and host will ultimately increase our fundamental understanding of pathogen biology, mechanisms responsible for the development of protective immunity, and guide next-generation vaccine design.
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                Author and article information

                Contributors
                Abbas Doosti: abbasdoosti@yahoo.com
                Journal
                Journal ID (nlm-ta): Arch Microbiol
                Journal ID (iso-abbrev): Arch Microbiol
                Title: Archives of Microbiology
                Publisher: Springer Berlin Heidelberg (Berlin/Heidelberg )
                ISSN (Print): 0302-8933
                ISSN (Electronic): 1432-072X
                Publication date (Electronic): 20 March 2023
                Publication date (Print): 2023
                Volume: 205
                Issue: 4
                Electronic Location Identifier: 122
                Affiliations
                [1 ]GRID grid.467523.1, ISNI 0000 0004 0493 9277, Department of Biology, Shahrekord Branch, , Islamic Azad University, ; Shahrekord, Iran
                [2 ]GRID grid.468149.6, ISNI 0000 0004 5907 0003, Biotechnology Research Center, Shahrekord Branch, , Islamic Azad University, ; Shahrekord, Iran
                [3 ]GRID grid.440801.9, ISNI 0000 0004 0384 8883, Cellular and Molecular Research Center, , Basic Health Sciences Research Institute, Shahrekord University of Medical Sciences, ; Shahrekord, Iran
                Author notes

                Communicated by Erko Stackebrandt.

                Article
                Publisher ID: 3471
                DOI: 10.1007/s00203-023-03471-6
                PMC ID: 10025791
                PubMed ID: 36939918
                SO-VID: 83f954d3-687d-4203-a2c2-32ea21872266
                Copyright © © The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature 2023, Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
                License:

                This article is made available via the PMC Open Access Subset for unrestricted research re-use and secondary analysis in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the World Health Organization (WHO) declaration of COVID-19 as a global pandemic.

                History
                Date received : 4 January 2023
                Date revision received : 25 February 2023
                Date accepted : 7 March 2023
                Categories
                Subject: Original Paper
                Custom metadata
                issue-copyright-statement © Springer-Verlag GmbH Germany, part of Springer Nature 2023

                ScienceOpen disciplines: Microbiology & Virology
                Keywords: brucella,lactococcuslactis,bls protein,mucosal live vector
                Data availability:
                ScienceOpen disciplines: Microbiology & Virology
                Keywords: brucella, lactococcuslactis, bls protein, mucosal live vector

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