Efficient transduction of murine primary T cells requires a combination of high viral titer, preferred tropism, and proper timing of transduction

Tong Zhang, Tom C. Tsang, David T. Harris

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Retroviral vectors have been used exclusively for genetic modification of primary T cells. Most T cell infection protocols have been developed for human T cells, whereas systematic investigations of the optimal conditions for transduction of murine primary T cells are limited. In this study, ecotropic and 10A1-pseudotyped retroviral vectors were compared for their efficiency in infecting murine primary T cell cells, as well as T cell lines. Various factors that affect transduction efficiency were also explored, including virus titer, times of exposure, timing of infection, low-speed centrifugation, and use of fibronectin fragment. Our results showed that up to 80% of murine primary T cells could be infected after a single exposure. Successful infection required a combination of high virus titer (>107 CFU/ml), proper timing of infection (within 24 h after mitogen stimulation), and preferred tropism (ecotropic vectors). These optimization results may help to establish a standard protocol for infection of murine primary T cells and provide some insight into the obstacles to retroviral infection of T cells.

Original languageEnglish (US)
Pages (from-to)123-130
Number of pages8
JournalJournal of Hematotherapy and Stem Cell Research
Volume12
Issue number1
DOIs
StatePublished - Feb 2003

Fingerprint

Tropism
T-Lymphocytes
Infection
Viral Load
Centrifugation
Mitogens
Fibronectins
Cell Line

ASJC Scopus subject areas

  • Hematology
  • Immunology

Cite this

@article{147dbf9e184843aeb931668e6c72c515,
title = "Efficient transduction of murine primary T cells requires a combination of high viral titer, preferred tropism, and proper timing of transduction",
abstract = "Retroviral vectors have been used exclusively for genetic modification of primary T cells. Most T cell infection protocols have been developed for human T cells, whereas systematic investigations of the optimal conditions for transduction of murine primary T cells are limited. In this study, ecotropic and 10A1-pseudotyped retroviral vectors were compared for their efficiency in infecting murine primary T cell cells, as well as T cell lines. Various factors that affect transduction efficiency were also explored, including virus titer, times of exposure, timing of infection, low-speed centrifugation, and use of fibronectin fragment. Our results showed that up to 80{\%} of murine primary T cells could be infected after a single exposure. Successful infection required a combination of high virus titer (>107 CFU/ml), proper timing of infection (within 24 h after mitogen stimulation), and preferred tropism (ecotropic vectors). These optimization results may help to establish a standard protocol for infection of murine primary T cells and provide some insight into the obstacles to retroviral infection of T cells.",
author = "Tong Zhang and Tsang, {Tom C.} and Harris, {David T.}",
year = "2003",
month = "2",
doi = "10.1089/152581603321210208",
language = "English (US)",
volume = "12",
pages = "123--130",
journal = "Stem Cells and Development",
issn = "1547-3287",
publisher = "Mary Ann Liebert Inc.",
number = "1",

}

TY - JOUR

T1 - Efficient transduction of murine primary T cells requires a combination of high viral titer, preferred tropism, and proper timing of transduction

AU - Zhang, Tong

AU - Tsang, Tom C.

AU - Harris, David T.

PY - 2003/2

Y1 - 2003/2

N2 - Retroviral vectors have been used exclusively for genetic modification of primary T cells. Most T cell infection protocols have been developed for human T cells, whereas systematic investigations of the optimal conditions for transduction of murine primary T cells are limited. In this study, ecotropic and 10A1-pseudotyped retroviral vectors were compared for their efficiency in infecting murine primary T cell cells, as well as T cell lines. Various factors that affect transduction efficiency were also explored, including virus titer, times of exposure, timing of infection, low-speed centrifugation, and use of fibronectin fragment. Our results showed that up to 80% of murine primary T cells could be infected after a single exposure. Successful infection required a combination of high virus titer (>107 CFU/ml), proper timing of infection (within 24 h after mitogen stimulation), and preferred tropism (ecotropic vectors). These optimization results may help to establish a standard protocol for infection of murine primary T cells and provide some insight into the obstacles to retroviral infection of T cells.

AB - Retroviral vectors have been used exclusively for genetic modification of primary T cells. Most T cell infection protocols have been developed for human T cells, whereas systematic investigations of the optimal conditions for transduction of murine primary T cells are limited. In this study, ecotropic and 10A1-pseudotyped retroviral vectors were compared for their efficiency in infecting murine primary T cell cells, as well as T cell lines. Various factors that affect transduction efficiency were also explored, including virus titer, times of exposure, timing of infection, low-speed centrifugation, and use of fibronectin fragment. Our results showed that up to 80% of murine primary T cells could be infected after a single exposure. Successful infection required a combination of high virus titer (>107 CFU/ml), proper timing of infection (within 24 h after mitogen stimulation), and preferred tropism (ecotropic vectors). These optimization results may help to establish a standard protocol for infection of murine primary T cells and provide some insight into the obstacles to retroviral infection of T cells.

UR - http://www.scopus.com/inward/record.url?scp=0037295242&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0037295242&partnerID=8YFLogxK

U2 - 10.1089/152581603321210208

DO - 10.1089/152581603321210208

M3 - Article

C2 - 12662443

AN - SCOPUS:0037295242

VL - 12

SP - 123

EP - 130

JO - Stem Cells and Development

JF - Stem Cells and Development

SN - 1547-3287

IS - 1

ER -