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      Virulence-associated trimeric autotransporters of Haemophilus parasuis are antigenic proteins expressed in vivo

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          Abstract

          Glässer’s disease is a re-emerging swine disease characterized by a severe septicaemia. Vaccination has been widely used to control the disease, although there is a lack of extended cross-protection. Trimeric autotransporters, a family of surface exposed proteins implicated in host-pathogen interactions, are good vaccine candidates. Members of this family have been described in Haemophilus parasuis and designated as virulence-associated trimeric autotransporters (VtaA). In this work, we produced 15 recombinant VtaA passenger domains and looked for the presence of antibodies directed against them in immune sera by immunoblotting. After infection with a subclinical dose of H. parasuis Nagasaki, an IgG mediated antibody response against 6 (VtaA1, 5, 6, 8, 9 and 10) of the 13 VtaA of the Nagasaki strain was detected, indicating that they are expressed in vivo. IgA production against VtaA was detected in only one animal. VtaA were more likely to be late antigens when compared to early (Omp P5 and Omp P6) and late (YaeT) defined antigens. Antibody cross-reaction with two orthologs of Nagasaki’s VtaA5 and 6, VtaA15 and 16 of strain HP1319, was also detected. No antibodies against VtaA were detected in the sera of animals immunized with a bacterin of the Nagasaki strain, suggesting poor expression in the in vitro conditions used. Taken together, these results indicate that VtaA are good candidate immunogens that could be used to improve H. parasuis vaccines. However, their capacity to confer protective immunity needs to be further studied.

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          NadA, a Novel Vaccine Candidate of Neisseria meningitidis

          Neisseria meningitidis is a human pathogen, which, in spite of antibiotic therapy, is still a major cause of mortality due to sepsis and meningitis. Here we describe NadA, a novel surface antigen of N. meningitidis that is present in 52 out of 53 strains of hypervirulent lineages electrophoretic types (ET) ET37, ET5, and cluster A4. The gene is absent in the hypervirulent lineage III, in N. gonorrhoeae and in the commensal species N. lactamica and N. cinerea. The guanine/cytosine content, lower than the chromosome, suggests acquisition by horizontal gene transfer and subsequent limited evolution to generate three well-conserved alleles. NadA has a predicted molecular structure strikingly similar to a novel class of adhesins (YadA and UspA2), forms high molecular weight oligomers, and binds to epithelial cells in vitro supporting the hypothesis that NadA is important for host cell interaction. NadA induces strong bactericidal antibodies and is protective in the infant rat model suggesting that this protein may represent a novel antigen for a vaccine able to control meningococcal disease caused by three hypervirulent lineages.
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            Trimeric autotransporters: a distinct subfamily of autotransporter proteins.

            Autotransporter proteins are a large family of gram-negative bacterial extracellular proteins. These proteins have a characteristic arrangement of functional domains, including an N-terminal signal peptide, an internal passenger domain, and a C-terminal translocator domain. Recent studies have identified a novel subfamily of autotransporters, defined by a short trimeric C-terminal translocator domain and known as trimeric autotransporters. In this article, we review our current knowledge of the structural and functional characteristics of trimeric autotransporters, highlighting the distinctions between this subfamily and conventional autotransporters. We speculate that trimeric autotransporters evolved to enable high-affinity multivalent adhesive interactions with host surfaces and circulating host molecules to take place.
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              Domain annotation of trimeric autotransporter adhesins—daTAA

              Motivation: Trimeric autotransporter adhesins (TAAs), such as Yersinia YadA, Neisseria NadA, Moraxella UspAs, Haemophilus Hia and Bartonella BadA, are important pathogenicity factors of proteobacteria. Their high sequence diversity and distinct mosaic-like structure lead to difficulties in the annotation of their sequences. These stem from the large number of short repeats, the presence of compositionally unusual coiled-coils, fuzzy domain boundaries and regions of seemingly low sequence complexity. Results: We have developed a workflow, named daTAA, for the accurate domain annotation of TAAs. Its core consists of manually curated alignments and of knowledge-based rules that enhance assignments made by sequence similarity. Compared to general domain annotation servers such as PFAM, daTAA captures more domains and provides more sensitive domain detection, as well as integrated and detailed coiled-coil assignments. Availability: The daTAA server is freely accessible at http://toolkit.tuebingen.mpg.de/dataa Contact: andrei.lupas@tuebingen.mpg.de Supplementary information: Supplementary data are available at Bioinformatics online.
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                Author and article information

                Journal
                Vet Res
                vetres
                Veterinary Research
                EDP Sciences
                0928-4249
                1297-9716
                10 December 2009
                May-Jun 2010
                10 December 2009
                : 41
                : 3 ( publisher-idID: vetres/2010/03 )
                : 26
                Affiliations
                [1 ] Centre de Recerca en Sanitat Animal (CReSA), UAB-IRTA, Campus de la Universitat Autònoma de Barcelona – Edifici CReSA 08193 Bellaterra, Barcelona Spain
                [2 ] Institut de Recerca i Tecnologia Agroalimentàries (IRTA) Barcelona Spain
                [3 ] HIPRA, S.A. Amer Girona Spain
                [4 ]Present address: Minnesota Veterinary Diagnostic Laboratory, College of Veterinary Medicine, University of Minnesota, St. Paul MN USA
                Author notes
                [* ]Corresponding author: alex.olvera@ 123456cresa.uab.cat
                [**]

                Both authors have equally contributed to the work.

                Article
                v09511 10.1051/vetres/2009074
                10.1051/vetres/2009074
                2820231
                19995512
                71a45f9f-0bda-4658-b9f4-fb45490adfcf
                © INRA, EDP Sciences, 2010

                This is an Open Access article distributed under the terms of the Creative Commons Attribution-Noncommercial License ( http://creativecommons.org/licenses/by-nc/3.0/), which permits unrestricted use, distribution, and reproduction in any noncommercial medium, provided the original work is properly cited.

                History
                : 09 September 2009
                : 07 December 2009
                Page count
                Figures: 5, Tables: 1, Equations: 0, References: 35, Pages: 11
                Categories
                Original Article

                Veterinary medicine
                vtaa,omp,antibody,haemophilus parasuis,cross-reactivity
                Veterinary medicine
                vtaa, omp, antibody, haemophilus parasuis, cross-reactivity

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