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      Utility of dominant epitopes derived from cell-wall protein LppZ for immunodiagnostic of pulmonary tuberculosis

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          Abstract

          Background

          Serological antibodies tests for tuberculosis (TB) are widely used in developing countries. They appear to have some advantages- faster, simple and could be used for extrapulmonary TB. However, most of current commercial TB serological tests are failed to provide sufficient sensitivity and specificity. Improved serological biomarkers were essential. In this study, we present an approach using peptide array to discover new immunodiagnostic biomarkers based on immunodominant epitopes of TB antigens.

          Results

          The Probable conserved lipoprotein LppZ, which is difficult to express and purify in vivo was selected as the model antigen. We use two-step screening for dominant epitope selection. Based on peptide array data from 170 TB patients and 41 control samples, two dominant epitopes were identified to have diagnostic value for TB patients. Truncation assay was used to identify the core reactive sequence. Peptide- based ELISA was used to evaluate the diagnostic ability of pep-LppZ-1 and pep-LppZ-13. Pep-LppZ-1 has a sensitivity of 49.2% and a specificity of 83.3% in TB diagnose. Pep-LppZ-13 has a sensitivity of 43.3% and a specificity of 88.5% in TB diagnose.

          Conclusions

          Our result demonstrated that peptide array screening would be an advantage strategy of screening TB diagnostic peptides.

          Electronic supplementary material

          The online version of this article (10.1186/s12865-018-0243-2) contains supplementary material, which is available to authorized users.

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          Most cited references21

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          Scalable high-density peptide arrays for comprehensive health monitoring.

          Joseph Legutki, Zhan-Gong Zhao, Matt Greving … (2014)
          There is an increasing awareness that health care must move from post-symptomatic treatment to presymptomatic intervention. An ideal system would allow regular inexpensive monitoring of health status using circulating antibodies to report on health fluctuations. Recently, we demonstrated that peptide microarrays can do this through antibody signatures (immunosignatures). Unfortunately, printed microarrays are not scalable. Here we demonstrate a platform based on fabricating microarrays (~10 M peptides per slide, 330,000 peptides per assay) on silicon wafers using equipment common to semiconductor manufacturing. The potential of these microarrays for comprehensive health monitoring is verified through the simultaneous detection and classification of six different infectious diseases and six different cancers. Besides diagnostics, these high-density peptide chips have numerous other applications both in health care and elsewhere.
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            Using Phage and Yeast Display to Select Hundreds of Monoclonal Antibodies: Application to Antigen 85, a Tuberculosis Biomarker

            Fortunato Ferrara, Leslie Naranjo, Sandeep Kumar-Barodia … (2012)
            Background Current diagnostic methods for tuberculosis (TB), a major global health challenge that kills nearly two million people annually, are time-consuming and inadequate. During infection a number of bacterial molecules that play a role in the infective process are released and have been proposed as biomarkers for early TB diagnosis. Antigen 85 (Ag85) is the most abundant secreted TB protein, and a potential target for this diagnostic approach. One of the bottlenecks in the direct detection of such bacterial targets is the availability of robust, sensitive, specific antibodies. Methods Using Ag85 as a model, we describe a method to select antibodies against any potential target using a novel combination of phage and yeast display that exploits the advantage of each approach. Results The efficiency of this approach was attested to by the 111 specific antibodies identified in initial screens. These were assessed for binding to the different Ag85 subunits, affinity, and activity in sandwich assays. Conclusions The novelty of this approach lies in the possibility of screening the entire output of a phage antibody selection in a single experiment by yeast display. This can be considered analogous to carrying out a million ELISAs. The monoclonal antibodies (mAbs) identified in this way show high binding affinity and selectivity for the antigens and offer an advantage over traditional mAbs produced by relatively expensive and time consuming techniques. This approach has wide applicability, and the affinity of selected antibodies can be significantly improved, if required.
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              Disease state differentiation and identification of tuberculosis biomarkers via native antigen array profiling.

              Mark Sartain, Richard Slayden, Krishna K. Singh … (2006)
              A critical element of tuberculosis control is early and sensitive diagnosis of infection and disease. Our laboratories recently showed that different stages of disease were distinguishable via two-dimensional Western blot analyses of Mycobacterium tuberculosis culture filtrate proteins. However, this methodology is not suitable for high throughput testing. Advances in protein microarray technology provide a realistic mechanism to screen a large number of serum samples against thousands of proteins to identify biomarkers of disease states. Techniques were established for separation of native M. tuberculosis cytosol and culture filtrate proteins, resulting in 960 unique protein fractions that were used to generate protein microarrays. Evaluation of serological reactivity from 42 patients in three tuberculosis disease states and healthy purified protein derivative-positive individuals demonstrated that human immunodeficiency virus (HIV)-negative cavitary and noncavitary tuberculosis (TB) patients' sera recognized 126 and 59 fractions, respectively. Sera from HIV patients coinfected with TB recognized 20 fractions of which five overlapped with those recognized by non-HIV TB patients' sera and 15 were unique to the HIV+TB+ disease state. Identification of antigens within the reactive fractions yielded 11 products recognized by both cavitary and noncavitary TB patients' sera and four proteins (HspX, MPT64, PstS1, and TrxC) specific to cavitary TB patients. Moreover four novel B cell antigens (BfrB, LppZ, SodC, and TrxC) of human tuberculosis were identified.
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                Author and article information

                Contributors
                Hairong Huang: Huanghairong@tb123.org
                Wentao Yue: yuewt@ccmu.edu.cn
                Journal
                Journal ID (nlm-ta): BMC Immunol
                Journal ID (iso-abbrev): BMC Immunol
                Title: BMC Immunology
                Publisher: BioMed Central (London )
                ISSN (Electronic): 1471-2172
                Publication date (Electronic): 1 March 2018
                Publication date PMC-release: 1 March 2018
                Publication date Collection: 2018
                Volume: 19
                Electronic Location Identifier: 10
                Affiliations
                [1 ]ISNI 0000 0004 1757 0026, GRID grid.414341.7, Department of Cellular and Molecular Biology, , Beijing Chest Hospital, Capital Medical University/Beijing Tuberculosis and Thoracic Tumor Research Institute, ; Beijing, 101149 China
                [2 ]ISNI 0000 0004 0369 153X, GRID grid.24696.3f, Department of Tuberculosis, , Beijing Chest Hospital, Capital Medical University, ; Beijing, 101149 China
                [3 ]ISNI 0000 0004 1757 0026, GRID grid.414341.7, National Clinical Laboratory on Tuberculosis, Beijing Key laboratory on Drug-resistant Tuberculosis Research, , Beijing Chest Hospital, Capital Medical University/Beijing Tuberculosis and Thoracic Tumor Institute, ; Beijing, 101149 China
                [4 ]ISNI 0000 0004 0369 153X, GRID grid.24696.3f, Central Laboratory, Beijing Obstetrics and Gynecology Hospital, , Capital Medical University, ; Chaoyang, Beijing, 100026 China
                Article
                Publisher ID: 243
                DOI: 10.1186/s12865-018-0243-2
                PMC ID: 5831716
                PubMed ID: 29490627
                SO-VID: d89248c2-7d8e-4391-98d3-020e6ab52531
                Copyright © © The Author(s). 2018
                License:

                Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. 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
                Date received : 12 July 2017
                Date accepted : 22 January 2018
                Funding
                Funded by: National Science and Technology Major Project of the Ministry of Science and Technology of China
                Categories
                Subject: Research Article
                Custom metadata
                issue-copyright-statement © The Author(s) 2018

                ScienceOpen disciplines: Immunology
                Keywords: tuberculosis peptide arrays,lppz,serologic test
                Data availability:
                ScienceOpen disciplines: Immunology
                Keywords: tuberculosis peptide arrays, lppz, serologic test

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