kat: A high-efficiency retroviral transduction system for primary human T lymphocytes

Mitchell H. Finer, Thomas J. Dull, Lu Qin, Deborah Parson, Margo R. Roberts

Research output: Contribution to journalArticle

Abstract

We describe a novel retroviral packaging system in which high titer amphotropic retrovirus was produced without the need to generate stable producer clones. kat expression vectors, which produce high levels of retroviral vector transcripts and retroviral packaging functions, were transfected into 293 cells followed by virus harvest 48 hours posttransfection. Viral titers as high as 3.8 proviral copies/cell/mL of frozen supernatant in 3T3 cells were obtained, 10- to 50-fold greater than transient viral titers reported using 3T3-based retroviral packaging lines. Cocultivation of primary human CD8+ T lymphocytes after transient transfection of 293 cells with kat plasmids resulted in transduction efficiencies of 10% to 40%, 5- to 10-fold greater compared to cocultivation with a high titer PA317 producer clone and significantly greater than previously reported results for transduction of primary human T lymphocytes with retroviral vectors. Virus produced using the kat system was shown to be free of detectable replication competent retrovirus by an extended provirus mobilization assay, demonstrating that this system is as safe as currently available stable packaging lines. The kat virus production system should be of general use for the rapid production of high titer viral supernatants, as well as for high-efficiency transduction hematopoietic cell types refractory to retroviral transduction.

Original languageEnglish (US)
Pages (from-to)43-50
Number of pages8
JournalBlood
Volume83
Issue number1
StatePublished - Jan 1 1994
Externally publishedYes

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T-cells
Product Packaging
Packaging
Viruses
T-Lymphocytes
Retroviridae
Coculture Techniques
Clone Cells
Proviruses
3T3 Cells
Refractory materials
Transfection
Assays
Plasmids

ASJC Scopus subject areas

  • Hematology

Cite this

Finer, M. H., Dull, T. J., Qin, L., Parson, D., & Roberts, M. R. (1994). kat: A high-efficiency retroviral transduction system for primary human T lymphocytes. Blood, 83(1), 43-50.

kat : A high-efficiency retroviral transduction system for primary human T lymphocytes. / Finer, Mitchell H.; Dull, Thomas J.; Qin, Lu; Parson, Deborah; Roberts, Margo R.

In: Blood, Vol. 83, No. 1, 01.01.1994, p. 43-50.

Research output: Contribution to journalArticle

Finer, MH, Dull, TJ, Qin, L, Parson, D & Roberts, MR 1994, 'kat: A high-efficiency retroviral transduction system for primary human T lymphocytes', Blood, vol. 83, no. 1, pp. 43-50.
Finer MH, Dull TJ, Qin L, Parson D, Roberts MR. kat: A high-efficiency retroviral transduction system for primary human T lymphocytes. Blood. 1994 Jan 1;83(1):43-50.
Finer, Mitchell H. ; Dull, Thomas J. ; Qin, Lu ; Parson, Deborah ; Roberts, Margo R. / kat : A high-efficiency retroviral transduction system for primary human T lymphocytes. In: Blood. 1994 ; Vol. 83, No. 1. pp. 43-50.
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