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      Application of M13 phage display for identifying immunogenic proteins from tick ( Ixodes scapularis) saliva

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          Abstract

          Background

          Ticks act as vectors for a large number of different pathogens, perhaps most notably Borrelia burgdorferi, the causative agent of Lyme disease. The most prominent tick vector in the United States is the blacklegged tick, Ixodes scapularis. Tick bites are of special public health concern since there are no vaccines available against most tick-transmitted pathogens. Based on the observation that certain non-natural host animals such as guinea pigs or humans can develop adaptive immune responses to tick bites, anti-tick vaccination is a potential approach to tackle health risks associated with tick bites.

          Results

          The aim of this study was to use an oligopeptide phage display strategy to identify immunogenic salivary gland proteins from I. scapularis that are recognized by human immune sera. Oligopeptide libraries were generated from salivary gland mRNA of 18 h fed nymphal I. scapularis. Eight immunogenic oligopeptides were selected using human immune sera. Three selected immunogenic oligopeptides were cloned and produced as recombinant proteins. The immunogenic character of an identified metalloprotease (MP1) was validated with human sera. This enzyme has been described previously and was hypothesized as immunogenic which was confirmed in this study. Interestingly, it also has close homologs in other Ixodes species.

          Conclusion

          An immunogenic protein of I. scapularis was identified by oligopeptide phage display. MP1 is a potential candidate for vaccine development.

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          Most cited references42

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          Blood-feeding arthropods: live syringes or invertebrate pharmacologists?

          J Ribeiro (1995)
          The habit of blood feeding evolved independently several times among the > 14,000 species and 400 genera of hematophagous arthropods. The specific need to remove blood from the host's skin led to sophisticated mechanical adaptations in invertebrate mouthparts. Moreover, the need to counteract the vertebrate host's hemostasis led to the evolution of salivary antihemostatic compounds injected into the host by these same mouthparts. The convergent evolution scenario for hematophagy has resulted in a large diversity of salivary anticlotting, antiplatelet, and vasodilatory substances. Thus, in addition to excelling as phlebotomists, hematophagous arthropods excel as pharmacologists.
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            A helper phage to improve single-chain antibody presentation in phage display.

            We show here that the number of single-chain antibody fragments (scFv) presented on filamentous phage particles generated with antibody display phagemids can be increased by more than two orders of magnitude by using a newly developed helper phage (hyperphage). Hyperphage have a wild-type pIII phenotype and are therefore able to infect F(+) Escherichia coli cells with high efficiency; however, their lack of a functional pIII gene means that the phagemid-encoded pIII-antibody fusion is the sole source of pIII in phage assembly. This results in an considerable increase in the fraction of phage particles carrying an antibody fragment on their surface. Antigen-binding activity was increased about 400-fold by enforced oligovalent antibody display on every phage particle. When used for packaging a universal human scFv library, hyperphage improved the specific enrichment factor obtained when panning on tetanus toxin. After two panning rounds, more than 50% of the phage were found to bind to the antigen, compared to 3% when conventional M13KO7 helper phage was used. Thus, hyperphage is particularly useful in stoichiometric situations, when there is little chance that a single phage will locate the desired antigen.
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              Host immunity to ticks.

              S K Wikel (1995)
              The tick-host-pathogen interface is characterized by complex immunological interactions. Tick feeding induces host immune regulatory and effector pathways involving antibodies, complement, antigen-presenting cells, T lymphocytes, and other bioactive molecules. Acquired resistance impairs tick engorgement, ova production, and viability. Tick countermeasures to host defenses reduce T-lymphocyte proliferation, elaboration of the TH1 cytokines interleukin-2 and interferon-gamma, production of macrophage cytokines interleukin-1 and tumor necrosis factor, and antibody responses. The dynamic balance between acquired resistance and tick modulation of host immunity affects engorgement and pathogen transmission. A thorough understanding of acquired immunity to ticks is essential for rational development of antitick vaccines.
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                Author and article information

                Contributors
                becker@ie-freiburg.mpg.de
                andre_felsberger@gmx.de
                andre.frenzel@tu-bs.de
                coywendy@hotmail.com
                mcd825@aol.com
                j.kuegler@tu-bs.de
                j.zantow@tu-bs.de
                tmather@mail.uri.edu
                m.hust@tu-bs.de
                Journal
                BMC Biotechnol
                BMC Biotechnol
                BMC Biotechnology
                BioMed Central (London )
                1472-6750
                30 May 2015
                30 May 2015
                2015
                : 15
                : 43
                Affiliations
                [ ]Institut für Biochemie, Biotechnologie und Bioinformatik, Technische Universität Braunschweig, Spielmannstr.7, 38106 Braunschweig, Germany
                [ ]University of Rhode Island, URI Center for Vector-Borne Disease, 231 Woodward Hall, 9 East Alumni Avenue, Suite 7, 02881 Kingston, RI USA
                [ ]Present Address: Max-Planck-Institute for Immunobiology and Epigenetics, Stuebeweg 51, 79108 Freiburg, Germany
                [ ]Present Address: YUMAB GmbH, Rebenring 33, 38106 Braunschweig, Germany
                Article
                167
                10.1186/s12896-015-0167-3
                4449557
                57312119-b9ac-4ddd-812b-63472ba49555
                © Becker et al.; licensee BioMed Central. 2015

                This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

                History
                : 20 January 2015
                : 15 May 2015
                Categories
                Research Article
                Custom metadata
                © The Author(s) 2015

                Biotechnology
                Biotechnology

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