Phenotypic Characterization of Chinese Rhesus Macaque Plasmablasts for Cloning Antigen-Specific Monoclonal Antibodies
Rhesus macaques ( ) are used as a human-relevant animal species for the evaluation of vaccines and as a source for cloning monoclonal antibodies (mAbs) that are highly similar to human-derived antibodies. Although antibody-secreting plasmablasts in humans are well-defined and can be easily isolated...
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| Published in | Frontiers in immunology Vol. 10; p. 2426 |
|---|---|
| Main Authors | , , , , , , , , , , , , , , , , |
| Format | Journal Article |
| Language | English |
| Published |
Switzerland
Frontiers Media S.A
11.10.2019
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| Abstract | Rhesus macaques (
) are used as a human-relevant animal species for the evaluation of vaccines and as a source for cloning monoclonal antibodies (mAbs) that are highly similar to human-derived antibodies. Although antibody-secreting plasmablasts in humans are well-defined and can be easily isolated for mAb cloning, it remains unclear whether the same phenotypic markers could be applied for isolating antibody-secreting plasmablasts from Chinese rhesus macaques. In this study, we evaluated a series of cell surface and intracellular markers and identified the phenotypic markers of plasmablasts in Chinese rhesus macaques as CD3
CD14
CD56
CD19
CD27
CD20
CD80
HLA-DR
CD95
. After influenza virus vaccination, the plasmablasts in peripheral blood mononuclear cells (PBMCs) increased transiently, peaked at day 4-7 after booster vaccination and returned to nearly undetectable levels by day 14. Antigen-specific enzyme-linked immunosorbent spot (ELISPOT) assays confirmed that the majority of the plasmablasts could produce influenza virus-specific antibodies. These plasmablasts showed transcriptional characteristics similar to those of human plasmablasts. Using single-cell PCR for immunoglobulin heavy and light chains, most mAbs cloned from the CD3
CD14
CD56
CD19
CD27
CD20
CD80
HLA-DR
CD95
plasmablasts after vaccination exhibited specific binding to influenza virus. This study defined the phenotypic markers for isolating antibody-secreting plasmablasts from Chinese rhesus macaques, which has implications for efficient cloning of mAbs and for the evaluation of plasmablast response after vaccination or infection in Chinese rhesus macaques. |
|---|---|
| AbstractList | Rhesus macaques (
) are used as a human-relevant animal species for the evaluation of vaccines and as a source for cloning monoclonal antibodies (mAbs) that are highly similar to human-derived antibodies. Although antibody-secreting plasmablasts in humans are well-defined and can be easily isolated for mAb cloning, it remains unclear whether the same phenotypic markers could be applied for isolating antibody-secreting plasmablasts from Chinese rhesus macaques. In this study, we evaluated a series of cell surface and intracellular markers and identified the phenotypic markers of plasmablasts in Chinese rhesus macaques as CD3
CD14
CD56
CD19
CD27
CD20
CD80
HLA-DR
CD95
. After influenza virus vaccination, the plasmablasts in peripheral blood mononuclear cells (PBMCs) increased transiently, peaked at day 4-7 after booster vaccination and returned to nearly undetectable levels by day 14. Antigen-specific enzyme-linked immunosorbent spot (ELISPOT) assays confirmed that the majority of the plasmablasts could produce influenza virus-specific antibodies. These plasmablasts showed transcriptional characteristics similar to those of human plasmablasts. Using single-cell PCR for immunoglobulin heavy and light chains, most mAbs cloned from the CD3
CD14
CD56
CD19
CD27
CD20
CD80
HLA-DR
CD95
plasmablasts after vaccination exhibited specific binding to influenza virus. This study defined the phenotypic markers for isolating antibody-secreting plasmablasts from Chinese rhesus macaques, which has implications for efficient cloning of mAbs and for the evaluation of plasmablast response after vaccination or infection in Chinese rhesus macaques. Rhesus macaques ( Macaca mulatta ) are used as a human-relevant animal species for the evaluation of vaccines and as a source for cloning monoclonal antibodies (mAbs) that are highly similar to human-derived antibodies. Although antibody-secreting plasmablasts in humans are well-defined and can be easily isolated for mAb cloning, it remains unclear whether the same phenotypic markers could be applied for isolating antibody-secreting plasmablasts from Chinese rhesus macaques. In this study, we evaluated a series of cell surface and intracellular markers and identified the phenotypic markers of plasmablasts in Chinese rhesus macaques as CD3 − CD14 − CD56 − CD19 − CD27 − CD20 −/low CD80 + HLA-DR + CD95 + . After influenza virus vaccination, the plasmablasts in peripheral blood mononuclear cells (PBMCs) increased transiently, peaked at day 4–7 after booster vaccination and returned to nearly undetectable levels by day 14. Antigen-specific enzyme-linked immunosorbent spot (ELISPOT) assays confirmed that the majority of the plasmablasts could produce influenza virus-specific antibodies. These plasmablasts showed transcriptional characteristics similar to those of human plasmablasts. Using single-cell PCR for immunoglobulin heavy and light chains, most mAbs cloned from the CD3 − CD14 − CD56 − CD19 − CD27 − CD20 −/low CD80 + HLA-DR + CD95 + plasmablasts after vaccination exhibited specific binding to influenza virus. This study defined the phenotypic markers for isolating antibody-secreting plasmablasts from Chinese rhesus macaques, which has implications for efficient cloning of mAbs and for the evaluation of plasmablast response after vaccination or infection in Chinese rhesus macaques. Rhesus macaques (Macaca mulatta) are used as a human-relevant animal species for the evaluation of vaccines and as a source for cloning monoclonal antibodies (mAbs) that are highly similar to human-derived antibodies. Although antibody-secreting plasmablasts in humans are well-defined and can be easily isolated for mAb cloning, it remains unclear whether the same phenotypic markers could be applied for isolating antibody-secreting plasmablasts from Chinese rhesus macaques. In this study, we evaluated a series of cell surface and intracellular markers and identified the phenotypic markers of plasmablasts in Chinese rhesus macaques as CD3−CD14−CD56−CD19−CD27−CD20−/lowCD80+HLA-DR+CD95+. After influenza virus vaccination, the plasmablasts in peripheral blood mononuclear cells (PBMCs) increased transiently, peaked at day 4–7 after booster vaccination and returned to nearly undetectable levels by day 14. Antigen-specific enzyme-linked immunosorbent spot (ELISPOT) assays confirmed that the majority of the plasmablasts could produce influenza virus-specific antibodies. These plasmablasts showed transcriptional characteristics similar to those of human plasmablasts. Using single-cell PCR for immunoglobulin heavy and light chains, most mAbs cloned from the CD3−CD14−CD56−CD19−CD27−CD20−/lowCD80+HLA-DR+CD95+ plasmablasts after vaccination exhibited specific binding to influenza virus. This study defined the phenotypic markers for isolating antibody-secreting plasmablasts from Chinese rhesus macaques, which has implications for efficient cloning of mAbs and for the evaluation of plasmablast response after vaccination or infection in Chinese rhesus macaques. |
| Author | Tan, Yee Joo Niu, Xuefeng Li, Pingchao He, Ping Pan, Weiqi Li, Jiashun Zhang, Fan Fan, Wenxia Luo, Jia Feng, Yupeng Li, Chufang Wang, Longyu Zheng, Zhiqiang Yang, Yanjia Liang, Renshan Yu, Haijian Chen, Ling |
| AuthorAffiliation | 3 State Key Laboratory of Respiratory Disease, First Affiliated Hospital of Guangzhou Medical University , Guangzhou , China 4 Department of Respiratory Medicine, Huadu People's Hospital , Guangzhou , China 1 State Key Laboratory of Respiratory Disease, Guangdong Laboratory of Computational Biomedicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences , Guangzhou , China 2 Institute of Physical Science and Information Technology, Anhui University , Hefei , China 6 Institute of Molecular and Cell Biology, A STAR (Agency for Science, Technology and Research) , Singapore , Singapore 5 Department of Microbiology and Immunology, Yong Loo Lin School of Medicine, National University Health System, National University of Singapore , Singapore , Singapore |
| AuthorAffiliation_xml | – name: 3 State Key Laboratory of Respiratory Disease, First Affiliated Hospital of Guangzhou Medical University , Guangzhou , China – name: 2 Institute of Physical Science and Information Technology, Anhui University , Hefei , China – name: 5 Department of Microbiology and Immunology, Yong Loo Lin School of Medicine, National University Health System, National University of Singapore , Singapore , Singapore – name: 6 Institute of Molecular and Cell Biology, A STAR (Agency for Science, Technology and Research) , Singapore , Singapore – name: 1 State Key Laboratory of Respiratory Disease, Guangdong Laboratory of Computational Biomedicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences , Guangzhou , China – name: 4 Department of Respiratory Medicine, Huadu People's Hospital , Guangzhou , China |
| Author_xml | – sequence: 1 givenname: Fan surname: Zhang fullname: Zhang, Fan organization: Institute of Physical Science and Information Technology, Anhui University, Hefei, China – sequence: 2 givenname: Longyu surname: Wang fullname: Wang, Longyu organization: Institute of Physical Science and Information Technology, Anhui University, Hefei, China – sequence: 3 givenname: Xuefeng surname: Niu fullname: Niu, Xuefeng organization: State Key Laboratory of Respiratory Disease, First Affiliated Hospital of Guangzhou Medical University, Guangzhou, China – sequence: 4 givenname: Jiashun surname: Li fullname: Li, Jiashun organization: Department of Respiratory Medicine, Huadu People's Hospital, Guangzhou, China – sequence: 5 givenname: Jia surname: Luo fullname: Luo, Jia organization: State Key Laboratory of Respiratory Disease, Guangdong Laboratory of Computational Biomedicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, China – sequence: 6 givenname: Yupeng surname: Feng fullname: Feng, Yupeng organization: State Key Laboratory of Respiratory Disease, Guangdong Laboratory of Computational Biomedicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, China – sequence: 7 givenname: Yanjia surname: Yang fullname: Yang, Yanjia organization: State Key Laboratory of Respiratory Disease, First Affiliated Hospital of Guangzhou Medical University, Guangzhou, China – sequence: 8 givenname: Ping surname: He fullname: He, Ping organization: State Key Laboratory of Respiratory Disease, Guangdong Laboratory of Computational Biomedicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, China – sequence: 9 givenname: Wenxia surname: Fan fullname: Fan, Wenxia organization: State Key Laboratory of Respiratory Disease, First Affiliated Hospital of Guangzhou Medical University, Guangzhou, China – sequence: 10 givenname: Renshan surname: Liang fullname: Liang, Renshan organization: State Key Laboratory of Respiratory Disease, First Affiliated Hospital of Guangzhou Medical University, Guangzhou, China – sequence: 11 givenname: Zhiqiang surname: Zheng fullname: Zheng, Zhiqiang organization: Institute of Molecular and Cell Biology, ASTAR (Agency for Science, Technology and Research), Singapore, Singapore – sequence: 12 givenname: Weiqi surname: Pan fullname: Pan, Weiqi organization: State Key Laboratory of Respiratory Disease, First Affiliated Hospital of Guangzhou Medical University, Guangzhou, China – sequence: 13 givenname: Chufang surname: Li fullname: Li, Chufang organization: State Key Laboratory of Respiratory Disease, First Affiliated Hospital of Guangzhou Medical University, Guangzhou, China – sequence: 14 givenname: Yee Joo surname: Tan fullname: Tan, Yee Joo organization: Institute of Molecular and Cell Biology, ASTAR (Agency for Science, Technology and Research), Singapore, Singapore – sequence: 15 givenname: Haijian surname: Yu fullname: Yu, Haijian organization: Department of Respiratory Medicine, Huadu People's Hospital, Guangzhou, China – sequence: 16 givenname: Ling surname: Chen fullname: Chen, Ling organization: State Key Laboratory of Respiratory Disease, First Affiliated Hospital of Guangzhou Medical University, Guangzhou, China – sequence: 17 givenname: Pingchao surname: Li fullname: Li, Pingchao organization: State Key Laboratory of Respiratory Disease, Guangdong Laboratory of Computational Biomedicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, China |
| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/31681312$$D View this record in MEDLINE/PubMed |
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| Copyright | Copyright © 2019 Zhang, Wang, Niu, Li, Luo, Feng, Yang, He, Fan, Liang, Zheng, Pan, Li, Tan, Yu, Chen and Li. Copyright © 2019 Zhang, Wang, Niu, Li, Luo, Feng, Yang, He, Fan, Liang, Zheng, Pan, Li, Tan, Yu, Chen and Li. 2019 Zhang, Wang, Niu, Li, Luo, Feng, Yang, He, Fan, Liang, Zheng, Pan, Li, Tan, Yu, Chen and Li |
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| Keywords | Chinese rhesus macaques influenza virus B cell plasmablast monoclonal antibodies vaccination |
| Language | English |
| License | Copyright © 2019 Zhang, Wang, Niu, Li, Luo, Feng, Yang, He, Fan, Liang, Zheng, Pan, Li, Tan, Yu, Chen and Li. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
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| Notes | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 Reviewed by: Hans-Martin Jäck, University of Erlangen Nuremberg, Germany; Leopoldo Flores-Romo, Center for Research and Advanced Studies (Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional), Mexico Edited by: Simone Cenci, San Raffaele Hospital (IRCCS), Italy This article was submitted to B Cell Biology, a section of the journal Frontiers in Immunology |
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) are used as a human-relevant animal species for the evaluation of vaccines and as a source for cloning monoclonal antibodies (mAbs) that... Rhesus macaques ( Macaca mulatta ) are used as a human-relevant animal species for the evaluation of vaccines and as a source for cloning monoclonal antibodies... Rhesus macaques (Macaca mulatta) are used as a human-relevant animal species for the evaluation of vaccines and as a source for cloning monoclonal antibodies... |
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| Title | Phenotypic Characterization of Chinese Rhesus Macaque Plasmablasts for Cloning Antigen-Specific Monoclonal Antibodies |
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