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      Phages Bearing Affinity Peptides to Bovine Rotavirus Differentiate the Virus from Other Viruses

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      Author(s): 1 , 2 , 3 , 1 , *
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      Publication date (Electronic):
      Journal: PLoS ONE
      Publisher: Public Library of Science

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          Abstract

          The aim of this study was to identify potential ligands and develop a novel diagnostic test to pathogenic bovine rotavirus (BRV) using phage display technology. The viruses were used as an immobilized target followed by incubation with a 12-mer phage display random peptide library. After five rounds of biopanning, phages had a specific binding activity to BRV were isolated. DNA sequencing indicated that phage displayed peptides HVHPPLRPHSDK, HATNHLPTPHNR or YPTHHAHTTPVR were potential ligands to BRV. Using the specific peptide-expressing phages, we developed a phage-based ELISA to differentiate BRV from other viruses. Compared with quantitative real-time PCR (qPCR), the phage-mediated ELISA was more suitable for the capture of BRV and the detection limitation of this approach was 0.1 µg/ml of samples. The high sensitivity, specificity and low cross-reactivity for the phage-based ELISA were confirmed in receiver operating characteristics (ROC) analysis.

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          Searching for peptide ligands with an epitope library.

          J Scott, G. Smith (1990)
          Tens of millions of short peptides can be easily surveyed for tight binding to an antibody, receptor or other binding protein using an "epitope library." The library is a vast mixture of filamentous phage clones, each displaying one peptide sequence on the virion surface. The survey is accomplished by using the binding protein to affinity-purify phage that display tight-binding peptides and propagating the purified phage in Escherichia coli. The amino acid sequences of the peptides displayed on the phage are then determined by sequencing the corresponding coding region in the viral DNA's. Potential applications of the epitope library include investigation of the specificity of antibodies and discovery of mimetic drug candidates.
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            Novel vaccine strategies to T-independent antigens.

            Gregory Lesinski, M.A.Julie Westerink (2001)
            T cell independent antigens do not require T cell help to induce an immune response, and are characterized by a lack of immunologic memory. These antigens can be divided into two classes, TI-1 or TI-2. TI-1 antigens, such as bacterial lipopolysaccharide, are potent B-cell mitogens, capable of non-specific, polyclonal activation of B cells. In contrast, TI-2 antigens can only activate mature B cells and consist of highly repetitive structures, such as capsular polysaccharides (CPS) from bacteria. Many vaccines currently in use consist of purified capsular polysaccharides from pathogenic bacteria such as Streptococcus pneumoniae and Neisseria meningitidis. These vaccines are efficacious in immune-competent adults, however, due to their TI-2 nature, are not effective in children <2 years of age. Converting polysaccharides into T cell dependent (TD) antigens, allows children, <2, to produce an effective immune response. This review focuses on various strategies used to convert the immune response to polysaccharide antigens from TI-2 to a TD response. Conjugate vaccines, anti-idiotypic antibodies, phage display library technology and DNA vaccines are discussed.
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              Action Mechanisms of Lithium Chloride on Cell Infection by Transmissible Gastroenteritis Coronavirus

              Xiaofeng Ren, Fandan Meng, Jiechao Yin … (2011)
              Transmissible gastroenteritis virus (TGEV) is a porcine coronavirus. Lithium chloride (LiCl) has been found to be effective against several DNA viruses, such as Herpes simplex virus and vaccinia virus. Recently, we and others have reported the inhibitory effect of LiCl on avian infectious bronchitis coronavirus (IBV) infection, an RNA virus. In the current study, the action mechanism of LiCl on cell infection by TGEV was investigated. Plaque assays and 3-(4,5)-dimethylthiahiazo(-z-y1)-3,5-di-phenyl tetrazoliumbromide (MTT) assays showed that the cell infection by TGEV was inhibited in a dose-dependent manner, when LiCl was added to virus-infected cells; the cell infection was not affected when either cells or viruses were pretreated with the drug. The inhibition of TGEV infection in vitro by LiCl was observed at different virus doses and with different cell lines. The inhibitory effect of LiCl against TGEV infection and transcription was confirmed by RT-PCR and real-time PCR targeting viral S and 3CL-protease genes. The time-of-addition effect of the drug on TGEV infection indicated that LiCl acted on the initial and late stage of TGEV infection. The production of virus was not detected at 36 h post-infection due to the drug treatment. Moreover, immunofluorescence (IF) and flow cytometry analyses based on staining of Annexin V and propidium iodide staining of nuclei indicated that early and late cell apoptosis induced by TGEV was inhibited efficiently. The ability of LiCl to inhibit apoptosis was investigated by IF analysis of caspase-3 expression. Our data indicate that LiCl inhibits TGEV infection by exerting an anti-apoptotic effect. The inhibitory effect of LiCl was also observed with porcine epidemic diarrhea coronavirus. Together with other reports concerning the inhibitory effect of lithium salts on IBV in cell culture, our results indicate that LiCl may be a potent agent against porcine and avian coronaviruses.
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                Author and article information

                Contributors
                : Role: Editor
                Journal
                Journal ID (nlm-ta): PLoS One
                Journal ID (publisher-id): plos
                Journal ID (pmc): plosone
                Title: PLoS ONE
                Publisher: Public Library of Science (San Francisco, USA )
                ISSN (Electronic): 1932-6203
                Publication date Collection: 2011
                Publication date (Electronic): 6 December 2011
                Volume: 6
                Issue: 12
                Electronic Location Identifier: e28667
                Affiliations
                [1 ]Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Northeast Agricultural University, Xiangfang District, Harbin, China
                [2 ]Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Northeast Agricultural University, Xiangfang District, Harbin, China
                [3 ]Department of Computer, College of Engineering, Northeast Agricultural University, Xiangfang District, Harbin, China
                Texas A&M, United States of America
                Author notes

                Conceived and designed the experiments: XR. Performed the experiments: XW GL XR. Analyzed the data: XW YR XR. Contributed reagents/materials/analysis tools: XR GL. Wrote the paper: XW XR.

                Article
                Publisher ID: PONE-D-11-18267
                DOI: 10.1371/journal.pone.0028667
                PMC ID: 3232237
                PubMed ID: 22163050
                SO-VID: 4d0c7dd1-c4e3-42d9-92f6-15e97be1f9b0
                Copyright statement: Wang et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
                History
                Date received : 15 September 2011
                Date accepted : 12 November 2011
                Page count
                Pages: 6
                Categories
                Subject: Research Article
                Subject: Biology
                Subject: Biotechnology
                Subject: Applied Microbiology
                Subject: Small Molecules
                Subject: Microbiology
                Subject: Virology
                Subject: Viral Disease Diagnosis
                Subject: Medicine
                Subject: Diagnostic Medicine
                Subject: Test Evaluation
                Subject: Veterinary Science
                Subject: Veterinary Diseases
                Subject: Veterinary Virology
                Subject: Veterinary Microbiology

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                ScienceOpen disciplines: Uncategorized

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