Cell cycle dependence of retroviral transduction: An issue of overlapping time scales

Abstract

Recombinant retroviruses are currently used as gene delivery vehicles for the purpose of gene therapy. It is generally believed that the efficiency of retroviral transduction depends on the cell cycle status of the target cells. However, it has been reported that this is not the case for the transduction of human and murine fibroblasts, in contrast to other cell types such as lymphocytes. The predictions of a mathematical model that we constructed, offer an explanation of this contradiction, based on the dynamics of the underlying processes of target cell growth and the intracellular decay of retroviral vectors. The model suggests that the utility of synchronization experiments, that are usually employed to study cell cycle specificity, is severely limited when the time scales of the above kinetic events are comparable to each other. The predictions of the model also suggest the use of retroviral vectors as cell cycle markers, as an alternative way to detect cell cycle dependence of retroviral transduction. This method obviates the need for cell synchronization and therefore, it does not perturb the cell cycle or interfere with the life cycle of retroviral vectors. Moreover, it does not depend on the intracellular stability of retroviral vectors. Our results show that in contrast to previously reported results, transduction of murine fibroblasts is cell cycle dependent, and they are consistent with the current notion that mitosis is the phase that confers transduction susceptibility. © 1998 John Wiley & Sons, Inc. Biotechnol Bioeng 58:272–281, 1998.

Tri-iodothyronine and a deleted form of hepatocyte growth factor act synergistically to enhance liver proliferation and enable in vivo retroviral gene transfer via the peripheral venous system

“…However, they require the target cells to be mitotically active for successful integration to occur. [5][6][7] The normal adult liver is mitotically quiescent and therefore resistant to transduction with (MoMLV) gene transfer. During the phase of hepatocyte proliferation retrovirus was administered via either the portal or tail vein.…”

Erratum in print version of "Toward a More Accurate Quantitation of the Activity of Recombinant Retroviruses: Alternatives to Titer and Multiplicity of Infection"

“…To be accurate, the number of CFU must be linearly proportional to cell number and our data show that this is true over a range of cell densities. However, this range can vary between cell types; as cell numbers were increased beyond this range, the number of CFU declined, possibly due to a decrease in the rate of cell growth, which is known to influence transduction (2,14). Equation 3 assumes that every particle that adsorbs on the cell surface will result in a successful gene transfer event; however, since the probability of each step after adsorption is Յ1 (parameter in equation 3), values of C vo are a lower limit.…”

Capture of antigen-specific T lymphocytes from human blood by selective immortalization to establish long-term T-cell lines maintaining primary cell characteristics☆

“…The strategy exploits a key property of the biology of murine leukemia virus (MuLV)-based retroviral vectors: they transduce only dividing cells [5][6][7][8][9]. Thus, transduction of cells from human PBMCs exposed to antigen should be effectively limited to those that respond and divide.…”