Reprogramming the antigen specificity of B cells using genome-editing technologies
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- James E Voss
- Department of Immunology and Microbiology, The Scripps Research Institute, La Jolla, United States
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- Alicia Gonzalez-Martin
- Department of Biochemistry, Universidad Autónoma de Madrid (UAM) and Instituto de Investigaciones Biomédicas Alberto Sols (CSIC-UAM), Madrid, Spain
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- Raiees Andrabi
- Department of Immunology and Microbiology, The Scripps Research Institute, La Jolla, United States
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- Roberta P Fuller
- Department of Immunology and Microbiology, The Scripps Research Institute, La Jolla, United States
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- Ben Murrell
- Department of Medicine, University of California, San Diego, San Diego, United States
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- Laura E McCoy
- Division of Infection and Immunity, University College London, London, United Kingdom
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- Katelyn Porter
- Department of Immunology and Microbiology, The Scripps Research Institute, La Jolla, United States
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- Deli Huang
- Department of Immunology and Microbiology, The Scripps Research Institute, La Jolla, United States
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- Wenjuan Li
- Department of Immunology and Microbiology, The Scripps Research Institute, La Jolla, United States
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- Devin Sok
- Department of Immunology and Microbiology, The Scripps Research Institute, La Jolla, United States
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- Khoa Le
- Department of Immunology and Microbiology, The Scripps Research Institute, La Jolla, United States
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- Bryan Briney
- Department of Immunology and Microbiology, The Scripps Research Institute, La Jolla, United States
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- Morgan Chateau
- Department of Molecular Microbiology and Immunology, Keck School of Medicine, University of Southern California, Los Angeles, United States
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- Geoffrey Rogers
- Department of Molecular Microbiology and Immunology, Keck School of Medicine, University of Southern California, Los Angeles, United States
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- Lars Hangartner
- Department of Immunology and Microbiology, The Scripps Research Institute, La Jolla, United States
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- Ann J Feeney
- Department of Immunology and Microbiology, The Scripps Research Institute, La Jolla, United States
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- David Nemazee
- Department of Immunology and Microbiology, The Scripps Research Institute, La Jolla, United States
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- Paula Cannon
- Department of Molecular Microbiology and Immunology, Keck School of Medicine, University of Southern California, Los Angeles, United States
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- Dennis R Burton
- Department of Immunology and Microbiology, The Scripps Research Institute, La Jolla, United States
抄録
<jats:p>We have developed a method to introduce novel paratopes into the human antibody repertoire by modifying the immunoglobulin (Ig) genes of mature B cells directly using genome editing technologies. We used CRISPR-Cas9 in a homology directed repair strategy, to replace the heavy chain (HC) variable region in B cell lines with that from an HIV broadly neutralizing antibody (bnAb), PG9. Our strategy is designed to function in cells that have undergone VDJ recombination using any combination of variable (V), diversity (D) and joining (J) genes. The modified locus expresses PG9 HC which pairs with native light chains (LCs) resulting in the cell surface expression of HIV specific B cell receptors (BCRs). Endogenous activation-induced cytidine deaminase (AID) in engineered cells allowed for Ig class switching and generated BCR variants with improved HIV neutralizing activity. Thus, BCRs engineered in this way retain the genetic flexibility normally required for affinity maturation during adaptive immune responses. Peripheral blood derived primary B cells from three different donors were edited using this strategy. Engineered cells could bind the PG9 epitope and sequenced mRNA showed PG9 HC transcribed as several different isotypes after culture with CD40 ligand and IL-4.</jats:p>
収録刊行物
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- eLife
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eLife 8 e42995-, 2019-01-17
eLife Sciences Publications, Ltd
