Selection of a Single Domain Antibody, Specific for an HLA-Bound Epitope of the Mycobacterial Ag85B Antigen
T cells recognizing epitopes on the surface of mycobacteria-infected macrophages can impart protection, but with associated risk for reactivation to lung pathology. We aimed to identify antibodies specific to such epitopes, which carry potentials for development toward novel therapeutic constructs....
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2020-10-01
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| Series: | Frontiers in Immunology |
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| Online Access: | https://www.frontiersin.org/article/10.3389/fimmu.2020.577815/full |
| _version_ | 1819054227164495872 |
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| author | Paola A. Ortega Mayra Silva-Miranda Mayra Silva-Miranda Alfredo Torres-Larios Eduardo Campos-Chávez Kees C. L. C. M. Franken Tom H. M. Ottenhoff Juraj Ivanyi Clara Espitia Clara Espitia |
| author_facet | Paola A. Ortega Mayra Silva-Miranda Mayra Silva-Miranda Alfredo Torres-Larios Eduardo Campos-Chávez Kees C. L. C. M. Franken Tom H. M. Ottenhoff Juraj Ivanyi Clara Espitia Clara Espitia |
| author_sort | Paola A. Ortega |
| collection | DOAJ |
| description | T cells recognizing epitopes on the surface of mycobacteria-infected macrophages can impart protection, but with associated risk for reactivation to lung pathology. We aimed to identify antibodies specific to such epitopes, which carry potentials for development toward novel therapeutic constructs. Since epitopes presented in the context of major histocompatibility complex alleles are rarely recognized by naturally produced antibodies, we used a phage display library for the identification of monoclonal human single domain antibody producing clones. The selected 2C clone displayed T cell receptor-like recognition of an HLA-A*0201 bound 199KLVANNTRL207 peptide from the Ag85B antigen, which is known to be an immunodominant epitope for human T cells. The specificity of the selected domain antibody was demonstrated by solid phase immunoassay and by immunofluorescent surface staining of peptide loaded cells of the T2 cell line. The antibody affinity binding was determined by biolayer interferometry. Our results validated the used technologies as suitable for the generation of antibodies against epitopes on the surface of Mycobacterium tuberculosis infected cells. The potential approaches forward the development of antibody in immunotherapy of tuberculosis have been outlined in the discussion. |
| first_indexed | 2024-12-21T12:48:16Z |
| format | Article |
| id | doaj.art-05ff3d91d2874bbf947b07cf67f0aee1 |
| institution | Directory Open Access Journal |
| issn | 1664-3224 |
| language | English |
| last_indexed | 2024-12-21T12:48:16Z |
| publishDate | 2020-10-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Immunology |
| spelling | doaj.art-05ff3d91d2874bbf947b07cf67f0aee12022-12-21T19:03:33ZengFrontiers Media S.A.Frontiers in Immunology1664-32242020-10-011110.3389/fimmu.2020.577815577815Selection of a Single Domain Antibody, Specific for an HLA-Bound Epitope of the Mycobacterial Ag85B AntigenPaola A. Ortega0Mayra Silva-Miranda1Mayra Silva-Miranda2Alfredo Torres-Larios3Eduardo Campos-Chávez4Kees C. L. C. M. Franken5Tom H. M. Ottenhoff6Juraj Ivanyi7Clara Espitia8Clara Espitia9Departamento de Inmunología, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, Ciudad de México, MéxicoDepartamento de Inmunología, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, Ciudad de México, MéxicoCONACyT-Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, Ciudad de México, MéxicoDepartment of Biochemistry and Structural Biology, Instituto de Fisiología Celular, Universidad Nacional Autónoma de México, Ciudad de México, MéxicoDepartment of Biochemistry and Structural Biology, Instituto de Fisiología Celular, Universidad Nacional Autónoma de México, Ciudad de México, MéxicoDepartment of Infectious Diseases, University Medical Centre Leiden, Leiden, NetherlandsDepartment of Infectious Diseases, University Medical Centre Leiden, Leiden, NetherlandsCenter for Host-Microbiome Interactions, King’s College London, London, United KingdomDepartamento de Inmunología, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, Ciudad de México, MéxicoCenter for Host-Microbiome Interactions, King’s College London, London, United KingdomT cells recognizing epitopes on the surface of mycobacteria-infected macrophages can impart protection, but with associated risk for reactivation to lung pathology. We aimed to identify antibodies specific to such epitopes, which carry potentials for development toward novel therapeutic constructs. Since epitopes presented in the context of major histocompatibility complex alleles are rarely recognized by naturally produced antibodies, we used a phage display library for the identification of monoclonal human single domain antibody producing clones. The selected 2C clone displayed T cell receptor-like recognition of an HLA-A*0201 bound 199KLVANNTRL207 peptide from the Ag85B antigen, which is known to be an immunodominant epitope for human T cells. The specificity of the selected domain antibody was demonstrated by solid phase immunoassay and by immunofluorescent surface staining of peptide loaded cells of the T2 cell line. The antibody affinity binding was determined by biolayer interferometry. Our results validated the used technologies as suitable for the generation of antibodies against epitopes on the surface of Mycobacterium tuberculosis infected cells. The potential approaches forward the development of antibody in immunotherapy of tuberculosis have been outlined in the discussion.https://www.frontiersin.org/article/10.3389/fimmu.2020.577815/fulltuberculosissingle domain antibodiespeptide-human leukocyte antigen complexmycobacterial Ag85BT cell receptor-like antibodies |
| spellingShingle | Paola A. Ortega Mayra Silva-Miranda Mayra Silva-Miranda Alfredo Torres-Larios Eduardo Campos-Chávez Kees C. L. C. M. Franken Tom H. M. Ottenhoff Juraj Ivanyi Clara Espitia Clara Espitia Selection of a Single Domain Antibody, Specific for an HLA-Bound Epitope of the Mycobacterial Ag85B Antigen Frontiers in Immunology tuberculosis single domain antibodies peptide-human leukocyte antigen complex mycobacterial Ag85B T cell receptor-like antibodies |
| title | Selection of a Single Domain Antibody, Specific for an HLA-Bound Epitope of the Mycobacterial Ag85B Antigen |
| title_full | Selection of a Single Domain Antibody, Specific for an HLA-Bound Epitope of the Mycobacterial Ag85B Antigen |
| title_fullStr | Selection of a Single Domain Antibody, Specific for an HLA-Bound Epitope of the Mycobacterial Ag85B Antigen |
| title_full_unstemmed | Selection of a Single Domain Antibody, Specific for an HLA-Bound Epitope of the Mycobacterial Ag85B Antigen |
| title_short | Selection of a Single Domain Antibody, Specific for an HLA-Bound Epitope of the Mycobacterial Ag85B Antigen |
| title_sort | selection of a single domain antibody specific for an hla bound epitope of the mycobacterial ag85b antigen |
| topic | tuberculosis single domain antibodies peptide-human leukocyte antigen complex mycobacterial Ag85B T cell receptor-like antibodies |
| url | https://www.frontiersin.org/article/10.3389/fimmu.2020.577815/full |
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