An optimized measles virus glycoprotein-pseudotyped lentiviral vector production system to promote efficient transduction of human primary B cells

Eirini Vamva; Stosh Ozog; Els Verhoeyen; Richard G. James; David J. Rawlings; Bruce E. Torbett

STAR Protocols (Mar 2022)

An optimized measles virus glycoprotein-pseudotyped lentiviral vector production system to promote efficient transduction of human primary B cells

Eirini Vamva,
Stosh Ozog,
Els Verhoeyen,
Richard G. James,
David J. Rawlings,
Bruce E. Torbett

Affiliations

Eirini Vamva: Department of Immunology and Microbiology, The Scripps Research Institute, La Jolla, CA, USA; Center for Immunity and Immunotherapies, Seattle Children’s Research Institute, Seattle, WA, USA; Department of Pediatrics, University of Washington School of Medicine, Seattle, WA, USA; Corresponding author
Stosh Ozog: Department of Immunology and Microbiology, The Scripps Research Institute, La Jolla, CA, USA; Department of Pediatrics, University of Washington School of Medicine, Seattle, WA, USA
Els Verhoeyen: CIRI–International Center for Infectiology Research, Team EVIR, Université de Lyon, Lyon, France
Richard G. James: Center for Immunity and Immunotherapies, Seattle Children’s Research Institute, Seattle, WA, USA; Department of Pediatrics, University of Washington School of Medicine, Seattle, WA, USA
David J. Rawlings: Center for Immunity and Immunotherapies, Seattle Children’s Research Institute, Seattle, WA, USA; Department of Pediatrics, University of Washington School of Medicine, Seattle, WA, USA
Bruce E. Torbett: Department of Immunology and Microbiology, The Scripps Research Institute, La Jolla, CA, USA; Center for Immunity and Immunotherapies, Seattle Children’s Research Institute, Seattle, WA, USA; Department of Pediatrics, University of Washington School of Medicine, Seattle, WA, USA; Institute for Stem Cell and Regenerative Medicine, Seattle, WA, USA; Corresponding author

Journal volume & issue: Vol. 3, no. 1
p. 101228

Abstract

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Summary: Measles virus envelope pseudotyped LV (MV-LV) can achieve high B cell transduction rates (up to 50%), but suffers from low titers. To overcome current limitations, we developed an optimized MV-LV production protocol that achieved consistent B cell transduction efficiency up to 75%. We detail this protocol along with analytical assays to assess the results of MV-LV mediated B cell transduction, including flow cytometry for B cell phenotypic characterization and measurement of transduction efficiency, and ddPCR for VCN analysis.

Published in STAR Protocols

ISSN: 2666-1667 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Science (General)
Website: https://star-protocols.cell.com/

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Abstract

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