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Generation of Multivirus-specific T Cells to Prevent/treat Viral Infections after Allogeneic Hematopoietic Stem Cell Transplant
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Retroviral transfer of T-cell receptor genes produces cells with a broad range of lytic activity

R J Orentas1, L A Bircher, S Roskopf

  • 1Department of Pediatrics, Section of Hematology-Oncology, Medical College of Wisconsin, Milwaukee, WI 53226, USA.

Scandinavian Journal of Immunology
|June 28, 2003
PubMed

The ability to transfer the T-cell receptor (TCR) for antigen using a retroviral vector has opened the door to a new paradigm for T-cell-based immunotherapy. Using recombinant TCRs, a population of activated T cells can now be redirected to recognize and lyse cellular targets according to the specificity afforded by the transduced TCR genes. To examine the range of lytic activity displayed by the transduced TCRs, transduced T cells were re-cloned by limiting dilution and quantitatively analysed for lytic activity. The lytic activity of the transduced TCRs varied considerably, as determined by the Km and Vmax of lysis. The lytic activity seen in the secondary clones generated from vector-transduced peripheral blood mononuclear cell demonstrated that one of the clones approached the lytic activity of the parental 'TCR donor' cytotoxic T-cell lymphocyte (CTL) clone, whereas the remainder demonstrated either reduced Vmax or reduced Vmax and Km. Thus, the lytic activity of a transduced TCR depends not only on its genetic sequence but also on the cellular context within which it is expressed. Analysis of TCR Vbeta transcript levels by real time polymerase chain reaction revealed that while total Vbeta gene expression was fairly constant, expression of the retrovirally transduced Vbeta chain varied greatly in transduced CD8+ CTL clones.

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