Retroviral immunotoxin gene therapy of acute myelogenous leukemia in mice using cytotoxic T cells transduced with an interleukin 4/diphtheria toxin gene.
The potential benefit of immunotoxin (IT) for cancer therapy has mostly been limited by the fact that only a small portion of injected dose ever reaches the cancer target. Thus, we set out to determine whether antigen-specific CTLs would be suitable vehicles to deliver IT to the site of cancer cells in vivo. A retroviral vector was constructed for gene therapy with (interleukin 4) IL-4 positioned downstream of its 20-amino-acid leader sequence that permitted cotranslational protein synthesis of IT along with truncated diphtheria toxin (DT). IL-4 was chosen as a ligand based on the expression of IL-4 receptor (IL-4R) on most acute myeloid leukemia cases. The first-time expression and secretion of a cytokine/DT fusion toxin was accomplished in mammalian NIH.3T3 cells, and then a retroviral vector was assembled. The retroviral IT was used to transiently transduce T15, a CD8+ CTL T cell line that specifically recognized C1498 (a lethal murine acute myeloid leukemia cell line). Transduced T15 T cells expressed intracellular DT and IL-4 as determined by immunofluorescence. Secreted IT supernatants collected from T15 had enzymatic activity and killed IL-4R+ C1498 cells, but not IL-4R- EL4 cells. Intravenous injection of transduced T15, but not nontransduced T15, into mice with s.c. tumors significantly inhibited tumor growth. In contrast, systemic therapy with a bacterial preparation of the same IL-4 IT given at its maximum tolerated dose did not protect. Retroviral IT-treated mice showed no sign of the renal or hepatic toxicity that is common to this class of IT. Together, these data indicate that retroviral IT may solve problems relating to systemic IT therapy by delivering reagent more directly to the site of cancer in vivo and may impart new anticancer defense mechanisms to antigen-specific T cells.
Vallera
D A
DA
University of Minnesota Cancer Center, Department of Therapeutic Radiology, Minneapolis 55455, USA. valle001@tc.umn.edu
Jin
N
N
Baldrica
J M
JM
Panoskaltsis-Mortari
A
A
Chen
S Y
SY
Blazar
B R
BR
D016475:3T3 Cells; D000818:Animals; D004167:Diphtheria Toxin / Q000235:genetics*; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D000922:Immunotoxins / Q000235:genetics*; D015847:Interleukin-4 / Q000235:genetics*; D007668:Kidney / Q000473:pathology; D015470:Leukemia, Myeloid, Acute / Q000628:therapy*; D008099:Liver / Q000473:pathology; D051379:Mice; D008810:Mice, Inbred C57BL; D012190:Retroviridae / Q000235:genetics*; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*
D004167:Diphtheria Toxin; D000922:Immunotoxins; D015847:Interleukin-4