Transduction of Human PBMC-Derived Dendritic Cells and Macrophages by an HIV-1-Based Lentiviral Vector System

Abstract

Professional antigen-presenting cells, such as dendritic cells (DCs) and macrophages, are target cells for gene therapy of infectious disease and cancer. However, transduction of DCs and macrophages has proved difficult by most currently available gene transfer methods. Several recent studies have shown that lentiviral vector systems can efficiently transduce many nondividing and differentiated cell types. In this study, we examined the gene transfer to DCs and macrophages using a lentiviral vector system. Human DCs were propagated from the adherent fraction of peripheral blood mononuclear cells (PBMCs) by culture in medium containing GM-CSF, IL-4, and TNF-alpha. Human macrophages were propagated from adherent PBMCs in medium containing GM-CSF. High titers of a replication-defective vesicular stomatitis virus glycoprotein G pseudotyped HIV-1-based vector encoding the enhanced yellow fluorescent protein were produced. In immature DCs (culture days 3 and 5), transduction efficiencies of 25 to 35% were achieved at a multiplicity of infection of 100. However, the transduction efficiency was decreased in more mature DCs (culture day 8 or later). Furthermore, monocyte-derived macrophages were also transduced by the lentiviral vector system. In addition, Alu-LTR PCR demonstrated the integration of the HIV-1 provirus into the cellular genome of the transduced DCs and macrophages. Allogeneic mixed lymphocyte reactions revealed similar antigen-presenting functions of untransduced and lentivirally transduced DCs. Thus, the results of this study demonstrate that both PBMC-derived DCs and macrophages can be transduced by lentiviral vectors.

Ex vivo generation of genetically modified dendritic cells for immunotherapy: implications of lymphocyte contamination

“…7,9,10,[31][32][33] We, and others, have shown the potential of HIV-1-based lentiviral vectors in delivering transgene to human monocyte-derived DCs and the induction of T-cell responses. 25,[34][35][36] The initial aim of the current study was to develop and optimize a clinically feasible method to generate gene-modified DC vaccines using an SIN lentiviral vector for cancer immunotherapy. Similar to previous findings, 37,38 the current study demonstrate that the SIN lentiviral vector could reproducibly transduce PB monocyte-derived immature DCs, resulting in the high level of transgene expression.…”

Lentiviral transduction of human T-lymphocytes with a RANTES intrakine inhibits human immunodeficiency virus type 1 infection

“…Such vectors have several compelling features: for example, they efficiently transduce either proliferating or quiescent cells and promote sustained expression of the transgene through their stable integration into the host cell genome. [17][18][19][20] In this study, we tested the feasibility of using lentiviral vectors for intrakine gene therapy. We employed a quantitative real-time PCR assay to determine the ability of intrakines to inhibit HIV-1 infection.…”

Dendritic cell-directed lentivector vaccine induces antigen-specific immune responses against murine melanoma

“…19 Several studies have shown that HIV-1-based lentivectors are able to deliver genes into non-dividing and less proliferating cells, including peripheral DCs. [20][21][22][23] One advantage of using the lentivector system is that the vector does not encode any major viral proteins. This eliminates the generation of competition of anti-vector responses, which competes with the generation of anti-transgene responses.…”