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Active immunization with Tocilizumab mimotopes induces specific immune responses

BACKGROUND: Tocilizumab is a humanized monoclonal antibody showing high-affinity binding to both soluble interleukin-6 receptor (sIL-6R) and membrane bound IL-6R (mIL-6R), thereby preventing pro-inflammatory effects of IL-6. However, therapeutic antibodies still have practical limitations. To overco...

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Autores principales: Yang, Lin, Xing, Rui, Li, Changhong, Liu, Yuan, Sun, Lin, Liu, Xiangyuan, Wang, Yongfu
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4450830/
https://www.ncbi.nlm.nih.gov/pubmed/26033236
http://dx.doi.org/10.1186/s12896-015-0161-9
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author Yang, Lin
Xing, Rui
Li, Changhong
Liu, Yuan
Sun, Lin
Liu, Xiangyuan
Wang, Yongfu
author_facet Yang, Lin
Xing, Rui
Li, Changhong
Liu, Yuan
Sun, Lin
Liu, Xiangyuan
Wang, Yongfu
author_sort Yang, Lin
collection PubMed
description BACKGROUND: Tocilizumab is a humanized monoclonal antibody showing high-affinity binding to both soluble interleukin-6 receptor (sIL-6R) and membrane bound IL-6R (mIL-6R), thereby preventing pro-inflammatory effects of IL-6. However, therapeutic antibodies still have practical limitations. To overcome these limitations, we generated Tocilizumab specific epitope mimics by using the phage display technology and tested whether the peptide mimics could induce similar humoral responses in mice immunized with the peptides. RESULTS: Seven phage mimics were obtained by using phage display peptide library. Four phage mimics (YHTTDKLFYMMR, YSAYEFEYILSS, KTMSAEEFDNWL and LTSHTYRSQADT) were shown to mimic Tocilizumab epitope using immunoassays. The mimotopes were conjugated to immunogenic carrier proteins and used to intraperitoneally immunize BALB/c mice. Sera from the mimotopes immunized mice not only showed specific binding to recombinant IL-6R, but can also IL-6R expressed in Hela, U-937, Jurkat cell lines and in fibroblast-like synoviocytes from patients with RA (FLS-RA). Furthermore, sera from mice immunized with mimotopes-KLH conjugate could reduce the level of phosphorylated- signal transducers and activator of transcription (STAT3), STAT3, phosphorylated- extracellular signal-regulated kinase (Erk) 1/2 and Erk1/2 in HeLa and Jurkat cells. Antibody-dependent cellular cytotoxicity (ADCC) assay showed that antibodies induced by mimotopes-KLH conjugate could elicit specific lysis in Hela and U-937 cells. CONCLUSIONS: From phage display library, we successfully isolated four Tocilizumab mimotopes which induced specific humoral and cellular reponses in vitro and in vivo.
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spelling pubmed-44508302015-06-02 Active immunization with Tocilizumab mimotopes induces specific immune responses Yang, Lin Xing, Rui Li, Changhong Liu, Yuan Sun, Lin Liu, Xiangyuan Wang, Yongfu BMC Biotechnol Research Article BACKGROUND: Tocilizumab is a humanized monoclonal antibody showing high-affinity binding to both soluble interleukin-6 receptor (sIL-6R) and membrane bound IL-6R (mIL-6R), thereby preventing pro-inflammatory effects of IL-6. However, therapeutic antibodies still have practical limitations. To overcome these limitations, we generated Tocilizumab specific epitope mimics by using the phage display technology and tested whether the peptide mimics could induce similar humoral responses in mice immunized with the peptides. RESULTS: Seven phage mimics were obtained by using phage display peptide library. Four phage mimics (YHTTDKLFYMMR, YSAYEFEYILSS, KTMSAEEFDNWL and LTSHTYRSQADT) were shown to mimic Tocilizumab epitope using immunoassays. The mimotopes were conjugated to immunogenic carrier proteins and used to intraperitoneally immunize BALB/c mice. Sera from the mimotopes immunized mice not only showed specific binding to recombinant IL-6R, but can also IL-6R expressed in Hela, U-937, Jurkat cell lines and in fibroblast-like synoviocytes from patients with RA (FLS-RA). Furthermore, sera from mice immunized with mimotopes-KLH conjugate could reduce the level of phosphorylated- signal transducers and activator of transcription (STAT3), STAT3, phosphorylated- extracellular signal-regulated kinase (Erk) 1/2 and Erk1/2 in HeLa and Jurkat cells. Antibody-dependent cellular cytotoxicity (ADCC) assay showed that antibodies induced by mimotopes-KLH conjugate could elicit specific lysis in Hela and U-937 cells. CONCLUSIONS: From phage display library, we successfully isolated four Tocilizumab mimotopes which induced specific humoral and cellular reponses in vitro and in vivo. BioMed Central 2015-06-02 /pmc/articles/PMC4450830/ /pubmed/26033236 http://dx.doi.org/10.1186/s12896-015-0161-9 Text en © Yang 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.
spellingShingle Research Article
Yang, Lin
Xing, Rui
Li, Changhong
Liu, Yuan
Sun, Lin
Liu, Xiangyuan
Wang, Yongfu
Active immunization with Tocilizumab mimotopes induces specific immune responses
title Active immunization with Tocilizumab mimotopes induces specific immune responses
title_full Active immunization with Tocilizumab mimotopes induces specific immune responses
title_fullStr Active immunization with Tocilizumab mimotopes induces specific immune responses
title_full_unstemmed Active immunization with Tocilizumab mimotopes induces specific immune responses
title_short Active immunization with Tocilizumab mimotopes induces specific immune responses
title_sort active immunization with tocilizumab mimotopes induces specific immune responses
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4450830/
https://www.ncbi.nlm.nih.gov/pubmed/26033236
http://dx.doi.org/10.1186/s12896-015-0161-9
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