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

被引：4
|
作者：
Vamva, Eirini [1 ,2 ,3 ]
Ozog, Stosh [1 ,3 ]
Verhoeyen, Els [4 ]
James, Richard G. [2 ,3 ]
Rawlings, David J. [2 ,3 ]
Torbett, Bruce E. [1 ,2 ,3 ,5 ]
机构：
[1] Scripps Res Inst, Dept Immunol & Microbiol, La Jolla, CA 92037 USA
[2] Seattle Childrens Res Inst, Ctr Immun & Immunotherapies, Seattle, WA 98121 USA
[3] Univ Washington, Sch Med, Dept Pediat, Seattle, WA 98195 USA
[4] Univ Lyon, CIRI Int Ctr Infectiol Res, Team EVIR, Lyon, France
[5] Inst Stem Cell & Regenerat Med, Seattle, WA 98109 USA
来源：
STAR PROTOCOLS | 2022年 / 3卷 / 01期
关键词：
Biotechnology and bioengineering; Cell isolation; Flow Cytometry/Mass Cytometry; Immunology; Microbiology; Molecular Biology;
D O I：
10.1016/j.xpro.2022.101228
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
Measles virus envelope pseudotyped LV (MV-LV) can achieve high B cell trans-duction rates (up to 50%), but suffers from low titers. To overcome current limi-tations, 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.
引用
收藏
页数：24
相关论文
共 50 条
  • [31] Improved transduction of human primitive hematopoietic cells with a lentiviral vector pseudotyped with the envelope protein of endogenous feline leukemia virus (RD114).
    Hanawa, H
    Kelly, PF
    Nathwani, AC
    Nienhuis, AW
    Vanin, EF
    BLOOD, 2000, 96 (11) : 524A - 524A
  • [32] Transduction of human hematopoietic stem cells by lentiviral vectors pseudotyped with the RD114-TR chimeric envelope glycoprotein
    Di Nunzio, F.
    Piovani, B.
    Cosset, F-L.
    Mavilio, F.
    Stornaiuolo, A.
    HUMAN GENE THERAPY, 2007, 18 (09) : 811 - 820
  • [33] A safe and efficient lentiviral vector system: Optimisation of virus production and purification
    Anson, DS
    MOLECULAR THERAPY, 2004, 9 : S27 - S27
  • [34] Efficient retrograde gene transfer into primate brain with an HIV-1-based lentiviral vector pseudotyped with rabies virus glycoprotein
    Inoue, Ken-ichi
    Kato, Shigeki
    Kobayashi, Kenta
    Miyachi, Shigehiro
    Shimada, Takashi
    Kobayashi, Kazuto
    Takada, Masahiko
    NEUROSCIENCE RESEARCH, 2008, 61 : S139 - S139
  • [35] Efficient transduction of nondividing human cells by feline immunodeficiency virus lentiviral vectors
    Eric M. Poeschla
    Flossie Wong-Staal
    David J. Looney
    Nature Medicine, 1998, 4 : 354 - 357
  • [36] Efficient transduction of nondividing human cells by feline immunodeficiency virus lentiviral vectors
    Poeschla, EM
    Wong-Staal, F
    Looney, DJ
    NATURE MEDICINE, 1998, 4 (03) : 354 - 357
  • [37] Efficient lentiviral transduction of primary human acute myelogenous and lymphoblastic leukemia cells
    Biagi, E
    Bambacioni, F
    Gaipa, G
    Casati, C
    Golay, J
    Biondi, A
    Introna, M
    HAEMATOLOGICA, 2001, 86 (01) : 13 - 16
  • [38] Transient blockade of TBK1/IKKε allows efficient transduction of primary human natural killer cells with vesicular stomatitis virus G-pseudotyped lentiviral vectors
    Chockley, Peter
    Patil, Sagar L.
    Gottschalk, Stephen
    CYTOTHERAPY, 2021, 23 (09) : 787 - 792
  • [39] EFFICIENT TRANSDUCTION OF T CELLS USING A SUSPENSION-BASED LENTIVIRAL VECTOR SYSTEM
    Parcelier, A.
    Larbi, A.
    Neveu, E.
    Cariou, A. Auffret
    Rousseaux, C.
    Charrier, S.
    CYTOTHERAPY, 2023, 25 (06) : S196 - S196
  • [40] Optimized lentiviral vector transduction of adherent cells and analysis in sulforhodamine B proliferation and chromatin immunoprecipitation assays
    Li, Luyuan
    Trent, Jonathan C.
    Eid, Josiane E.
    STAR PROTOCOLS, 2023, 4 (01):
12345