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Reprogramming the antigen specificity of B cells using genome-editing technologies.

James E Voss1,2,3, Alicia Gonzalez-Martin4, Raiees Andrabi1,2,3

  • 1Department of Immunology and Microbiology, The Scripps Research Institute, La Jolla, United States.

Elife
|January 17, 2019
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Summary

Scientists engineered human B cells to create novel antibodies targeting HIV. This genome editing method introduces specific antibody genes, enhancing the body's natural defense mechanisms against the virus.

Keywords:
B cellCAR-BHIVbnAbengineeringhumanimmunologyinfectious diseaseinflammationmicrobiologyvaccine

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Area of Science:

  • Immunology
  • Molecular Biology
  • Genetics

Background:

  • The human antibody repertoire is crucial for adaptive immunity.
  • Developing novel antibodies, particularly for challenging targets like HIV, remains a significant scientific pursuit.
  • Existing methods for antibody generation can be limited in scope and efficiency.

Purpose of the Study:

  • To develop a method for introducing novel antibody specificities into the human B cell repertoire.
  • To engineer B cells to express HIV-specific B cell receptors (BCRs) using genome editing.
  • To assess the functionality and diversity of engineered BCRs for potential therapeutic applications.

Main Methods:

  • Utilized CRISPR-Cas9 genome editing with homology-directed repair to replace immunoglobulin heavy chain variable regions in B cell lines.
  • Introduced the heavy chain variable region from an HIV broadly neutralizing antibody (bnAb), PG9.
  • Employed endogenous activation-induced cytidine deaminase (AID) for immunoglobulin class switching and affinity maturation.
  • Applied the strategy to peripheral blood-derived primary B cells from multiple donors.

Main Results:

  • Successfully engineered B cells expressed PG9 heavy chains paired with native light chains, leading to cell surface expression of HIV-specific BCRs.
  • Engineered BCRs demonstrated binding to the PG9 epitope.
  • Activation-induced cytidine deaminase facilitated Ig class switching, generating BCR variants with enhanced HIV neutralizing activity.
  • Primary B cells from different donors were successfully edited, expressing PG9 heavy chains as multiple isotypes after stimulation.

Conclusions:

  • Genome editing of mature B cells offers a viable strategy for introducing novel antibody specificities, such as those targeting HIV.
  • This approach retains the natural genetic flexibility of B cells for affinity maturation, leading to improved antibody function.
  • The engineered B cells and their expressed BCRs hold potential for developing new immunotherapies against HIV infection.