Human gene transfer

Characterization of human tumor-infiltrating lymphocytes as vehicles for retroviral-mediated gene transfer in man

Attan Kasid, Shoshana Morecki, Paul Aebersold, Kenneth Cornetta, Kenneth Culver, Scott Freeman, Elaine Director, Michael T. Lotze, R. Michael Blaese, W. French Anderson, Steven A. Rosenberg

Research output: Contribution to journalArticle

126 Citations (Scopus)

Abstract

Tumor-infiltrating lymphocytes (TILs) are cells generated from tumor suspensions cultured in interleukin 2 that can mediate cancer regression when adoptively transferred into mice or humans. Since TILs proliferate rapidly in vitro, recirculate, and preferentially localize at the tumor site in vivo, they provide an attractive model for delivery of exogenous genetic material into man. To determine whether efficient gene transfer into TILs is feasible, we transduced human TILs with the bacterial gene for neomycin-resistance (NeoR) using the retroviral vector N2. The transduced TIL populations were stable and polyclonal with respect to the intact NeoR gene integration and expressed high levels of neomycin phosphotransferase activity. The NeoR gene insertion did not alter the in vitro growth pattern and interleukin 2 dependence of the transduced TILs. Analyses of T-cell receptor gene rearrangement for β- and γ-chain genes revealed the oligoclonal nature of the TIL populations with no major change in the DNA rearrangement patterns or the levels of mRNA expression of the β and γ chains following transduction and selection of TILs in the neomycin analog G418. Human TILs expressed mRNA for tumor necrosis factors (α and β) and interleukin 2 receptor P55 but not for interleukin 1β, granulocyte/macrophage colony-stimulating factor, interleukin 6, and Interferon γ when grown with high-dose interleukin 2 without subsequent activation with mitogen or specific antigen. This pattern of cytokine-mRNA expression was not significantly altered following the transduction of TILs. The NeoR gene-transduced TILs could thus be used to follow the trafficking and survival of TILs in vivo, and clinical protocols using these transduced TILs in cancer patients have begun. The studies demonstrate the feasibility of TILs as suitable cellular vehicles for the introduction of therapeutic genes into patients receiving autologous TILs.

Original languageEnglish (US)
Pages (from-to)473-477
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume87
Issue number1
StatePublished - 1990
Externally publishedYes

Fingerprint

Tumor-Infiltrating Lymphocytes
Genes
Neomycin
Interleukin-2
Messenger RNA
Neoplasms
Tumor Necrosis Factors
T-Lymphocyte Gene Rearrangement
Kanamycin Kinase
T-Cell Receptor Genes
Bacterial Genes
Gene Rearrangement
Interleukin-2 Receptors
Insertional Mutagenesis
Population Dynamics
Feasibility Studies
Granulocyte-Macrophage Colony-Stimulating Factor
Clinical Protocols
Interleukin-1

Keywords

  • Gene therapy
  • Neomycin-resistance gene

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

Human gene transfer : Characterization of human tumor-infiltrating lymphocytes as vehicles for retroviral-mediated gene transfer in man. / Kasid, Attan; Morecki, Shoshana; Aebersold, Paul; Cornetta, Kenneth; Culver, Kenneth; Freeman, Scott; Director, Elaine; Lotze, Michael T.; Michael Blaese, R.; Anderson, W. French; Rosenberg, Steven A.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 87, No. 1, 1990, p. 473-477.

Research output: Contribution to journalArticle

Kasid, A, Morecki, S, Aebersold, P, Cornetta, K, Culver, K, Freeman, S, Director, E, Lotze, MT, Michael Blaese, R, Anderson, WF & Rosenberg, SA 1990, 'Human gene transfer: Characterization of human tumor-infiltrating lymphocytes as vehicles for retroviral-mediated gene transfer in man', Proceedings of the National Academy of Sciences of the United States of America, vol. 87, no. 1, pp. 473-477.
Kasid, Attan ; Morecki, Shoshana ; Aebersold, Paul ; Cornetta, Kenneth ; Culver, Kenneth ; Freeman, Scott ; Director, Elaine ; Lotze, Michael T. ; Michael Blaese, R. ; Anderson, W. French ; Rosenberg, Steven A. / Human gene transfer : Characterization of human tumor-infiltrating lymphocytes as vehicles for retroviral-mediated gene transfer in man. In: Proceedings of the National Academy of Sciences of the United States of America. 1990 ; Vol. 87, No. 1. pp. 473-477.
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