Reprogramming the antigen specificity of B cells using genome-editing technologies
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 reg...
| Main Authors: | , , , , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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eLife Sciences Publications Ltd
2019-01-01
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| Series: | eLife |
| Subjects: | |
| Online Access: | https://elifesciences.org/articles/42995 |
| id |
doaj-672ae969491c41729fd1ce5dbcadba72 |
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| record_format |
Article |
| collection |
DOAJ |
| language |
English |
| format |
Article |
| sources |
DOAJ |
| author |
James E Voss Alicia Gonzalez-Martin Raiees Andrabi Roberta P Fuller Ben Murrell Laura E McCoy Katelyn Porter Deli Huang Wenjuan Li Devin Sok Khoa Le Bryan Briney Morgan Chateau Geoffrey Rogers Lars Hangartner Ann J Feeney David Nemazee Paula Cannon Dennis R Burton |
| spellingShingle |
James E Voss Alicia Gonzalez-Martin Raiees Andrabi Roberta P Fuller Ben Murrell Laura E McCoy Katelyn Porter Deli Huang Wenjuan Li Devin Sok Khoa Le Bryan Briney Morgan Chateau Geoffrey Rogers Lars Hangartner Ann J Feeney David Nemazee Paula Cannon Dennis R Burton Reprogramming the antigen specificity of B cells using genome-editing technologies eLife CAR-B B cell engineering HIV bnAb vaccine |
| author_facet |
James E Voss Alicia Gonzalez-Martin Raiees Andrabi Roberta P Fuller Ben Murrell Laura E McCoy Katelyn Porter Deli Huang Wenjuan Li Devin Sok Khoa Le Bryan Briney Morgan Chateau Geoffrey Rogers Lars Hangartner Ann J Feeney David Nemazee Paula Cannon Dennis R Burton |
| author_sort |
James E Voss |
| title |
Reprogramming the antigen specificity of B cells using genome-editing technologies |
| title_short |
Reprogramming the antigen specificity of B cells using genome-editing technologies |
| title_full |
Reprogramming the antigen specificity of B cells using genome-editing technologies |
| title_fullStr |
Reprogramming the antigen specificity of B cells using genome-editing technologies |
| title_full_unstemmed |
Reprogramming the antigen specificity of B cells using genome-editing technologies |
| title_sort |
reprogramming the antigen specificity of b cells using genome-editing technologies |
| publisher |
eLife Sciences Publications Ltd |
| series |
eLife |
| issn |
2050-084X |
| publishDate |
2019-01-01 |
| description |
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. |
| topic |
CAR-B B cell engineering HIV bnAb vaccine |
| url |
https://elifesciences.org/articles/42995 |
| work_keys_str_mv |
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doaj-672ae969491c41729fd1ce5dbcadba722021-05-05T17:19:37ZengeLife Sciences Publications LtdeLife2050-084X2019-01-01810.7554/eLife.42995Reprogramming the antigen specificity of B cells using genome-editing technologiesJames E Voss0https://orcid.org/0000-0002-4777-1596Alicia Gonzalez-Martin1Raiees Andrabi2Roberta P Fuller3Ben Murrell4Laura E McCoy5Katelyn Porter6Deli Huang7Wenjuan Li8Devin Sok9Khoa Le10Bryan Briney11Morgan Chateau12Geoffrey Rogers13Lars Hangartner14Ann J Feeney15David Nemazee16Paula Cannon17Dennis R Burton18Department of Immunology and Microbiology, The Scripps Research Institute, La Jolla, United States; International AIDS Vaccine Initiative Neutralizing Antibody Center, The Scripps Research Institute, La Jolla, United States; Scripps Center for HIV/AIDS Vaccine Immunology and Immunogen Discovery (CHAVI-ID), The Scripps Research Institute, La Jolla, United StatesDepartment of Biochemistry, Universidad Autónoma de Madrid (UAM) and Instituto de Investigaciones Biomédicas Alberto Sols (CSIC-UAM), Madrid, SpainDepartment of Immunology and Microbiology, The Scripps Research Institute, La Jolla, United States; International AIDS Vaccine Initiative Neutralizing Antibody Center, The Scripps Research Institute, La Jolla, United States; Scripps Center for HIV/AIDS Vaccine Immunology and Immunogen Discovery (CHAVI-ID), The Scripps Research Institute, La Jolla, United StatesDepartment of Immunology and Microbiology, The Scripps Research Institute, La Jolla, United States; International AIDS Vaccine Initiative Neutralizing Antibody Center, The Scripps Research Institute, La Jolla, United States; Scripps Center for HIV/AIDS Vaccine Immunology and Immunogen Discovery (CHAVI-ID), The Scripps Research Institute, La Jolla, United StatesDepartment of Medicine, University of California, San Diego, San Diego, United States; Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, Stockholm, SwedenDivision of Infection and Immunity, University College London, London, United KingdomDepartment of Immunology and Microbiology, The Scripps Research Institute, La Jolla, United States; International AIDS Vaccine Initiative Neutralizing Antibody Center, The Scripps Research Institute, La Jolla, United States; Scripps Center for HIV/AIDS Vaccine Immunology and Immunogen Discovery (CHAVI-ID), The Scripps Research Institute, La Jolla, United StatesDepartment of Immunology and Microbiology, The Scripps Research Institute, La Jolla, United StatesDepartment of Immunology and Microbiology, The Scripps Research Institute, La Jolla, United StatesDepartment of Immunology and Microbiology, The Scripps Research Institute, La Jolla, United States; International AIDS Vaccine Initiative Neutralizing Antibody Center, The Scripps Research Institute, La Jolla, United States; Scripps Center for HIV/AIDS Vaccine Immunology and Immunogen Discovery (CHAVI-ID), The Scripps Research Institute, La Jolla, United StatesDepartment of Immunology and Microbiology, The Scripps Research Institute, La Jolla, United States; International AIDS Vaccine Initiative Neutralizing Antibody Center, The Scripps Research Institute, La Jolla, United States; Scripps Center for HIV/AIDS Vaccine Immunology and Immunogen Discovery (CHAVI-ID), The Scripps Research Institute, La Jolla, United StatesDepartment of Immunology and Microbiology, The Scripps Research Institute, La Jolla, United States; International AIDS Vaccine Initiative Neutralizing Antibody Center, The Scripps Research Institute, La Jolla, United States; Scripps Center for HIV/AIDS Vaccine Immunology and Immunogen Discovery (CHAVI-ID), The Scripps Research Institute, La Jolla, United StatesDepartment of Molecular Microbiology and Immunology, Keck School of Medicine, University of Southern California, Los Angeles, United StatesDepartment of Molecular Microbiology and Immunology, Keck School of Medicine, University of Southern California, Los Angeles, United StatesDepartment of Immunology and Microbiology, The Scripps Research Institute, La Jolla, United StatesDepartment of Immunology and Microbiology, The Scripps Research Institute, La Jolla, United StatesDepartment of Immunology and Microbiology, The Scripps Research Institute, La Jolla, United StatesDepartment of Molecular Microbiology and Immunology, Keck School of Medicine, University of Southern California, Los Angeles, United StatesDepartment of Immunology and Microbiology, The Scripps Research Institute, La Jolla, United States; International AIDS Vaccine Initiative Neutralizing Antibody Center, The Scripps Research Institute, La Jolla, United States; Scripps Center for HIV/AIDS Vaccine Immunology and Immunogen Discovery (CHAVI-ID), The Scripps Research Institute, La Jolla, United States; Ragon Institute of Massachusetts General Hospital, Massachusetts Institute of Technology, and Harvard, Cambridge, United StatesWe 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.https://elifesciences.org/articles/42995CAR-BB cellengineeringHIVbnAbvaccine |