Reprogramming the antigen specificity of B cells using genome-editing technologies
| Autor: | Lars Hangartner, Katelyn Porter, Raiees Andrabi, Deli Huang, Geoffrey L. Rogers, Wenjuan Li, Morgan Chateau, Devin Sok, Laura E. McCoy, Khoa Le, Dennis R. Burton, David Nemazee, Paula M. Cannon, Ben Murrell, Bryan Briney, Roberta P. Fuller, Alicia Gonzalez-Martin, Ann J. Feeney, James E. Voss |
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| Přispěvatelé: | National Institutes of Health (US), Bill & Melinda Gates Foundation, Ministerio de Ciencia, Innovación y Universidades (España), Ministerio de Economía y Competitividad (España), European Commission |
| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
engineering HIV Antibodies Epitope Antigen-Antibody Reactions 0302 clinical medicine Immunology and Inflammation Antibody Specificity vaccine Immunology and Allergy Biology (General) Gene Editing Microbiology and Infectious Disease B cell B-Lymphocytes biology General Neuroscience General Medicine Cytidine deaminase 3. Good health Cell biology medicine.anatomical_structure Medicine Antibody Immunoglobulin Heavy Chains CAR-B Human QH301-705.5 Science Immunology B-cell receptor Short Report Receptors Antigen B-Cell General Biochemistry Genetics and Molecular Biology Cell Line Affinity maturation 03 medical and health sciences Cytidine Deaminase bnAb medicine Humans CD40 General Immunology and Microbiology HIV Antibodies Neutralizing 030104 developmental biology Immunoglobulin class switching biology.protein CRISPR-Cas Systems 030217 neurology & neurosurgery |
| Zdroj: | eLife Digital.CSIC. Repositorio Institucional del CSIC instname eLife, Vol 8 (2019) |
| ISSN: | 2050-084X |
| Popis: | 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. This work was supported by the National Institutes of Health, R01-5R01DE025167-04, by The Bill and Melinda Gates Foundation OPP1183956, by CHAVI-ID grant UM1 AI100663, by the Ramon y Cajal Merit Award from Ministerio de Ciencia, Innovacion y Universidades, Spain (RYC-2016–21155 to AG-M), and by a Marie-Curie Fellowship (FP7-PEOPLE-2013-IOF to LEM). |
| Databáze: | OpenAIRE |
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