Brain engraftment of autologous macrophages transduced with a lentiviral flap vector: an approach to complement brain dysfunctions

Mordelet, Elodie; Kissa, Karima; Calvo, Charles‐Félix; Lebastard, M.; Milon, Geneviève; Werf, Sylvie van der; Vidal, Catherine; Charneau, Pierre

doi:10.1038/sj.gt.3301591

Cited by 15 publications

(18 citation statements)

References 42 publications

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“…Unfortunately, most classes of viral vector have proven to be poor agents to transfer genes into microglia and other myeloid lineages. One possible exception are vectors derived from complex lentiviruses, such as HIV, 35,36 which naturally target this cell type. However, most such vectors also carry significant stretches of native viral coding sequence, because their packaging signals are embedded within viral open reading frames.…”

Section: Discussionmentioning

confidence: 99%

Selective gene expression in brain microglia mediated via adeno-associated virus type 2 and type 5 vectors

et al. 2003

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Microglia represent a crucial cell population in the central nervous system, participating in the regulation and surveillance of physiological processes as well as playing key roles in the etiologies of several major brain disorders. The ability to target gene transfer vehicles selectively to microglia would provide a powerful new approach to investigations of mechanisms regulating brain pathologies, as well as enable the development of novel therapeutic strategies. In this study, we evaluate the feasibility of specifically and efficiently targeting microglia relative to other brain cells, using vectors based on two different serotypes of adeno-associated virus (AAV) carrying cell-type-specific transcriptional elements to regulate gene expression. Among a set of promoter choices examined, an element derived from the gene for the murine macrophage marker F4/80 was the most discriminating for microglia. Gene expression from vectors controlled by this element was highly selective for microglia, both in vitro and in vivo. To our knowledge, this is the first demonstration of selective expression of transferred genes in microglia using AAV-derived vectors, as well as the first utilization of recombinant AAV-5 vectors in any macrophage lineage. These results provide strong encouragement for the application of these vectors and this approach for delivering therapeutic and other genes selectively to microglia.

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Section: Discussionmentioning

confidence: 99%

Selective gene expression in brain microglia mediated via adeno-associated virus type 2 and type 5 vectors

et al. 2003

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“…Among the cell types which are susceptible to HIV-1-based vectors are hematopoietic stem cells [1][2][3][4][5][6][7] and monocytes/macrophages [8][9][10][11][12][13] which represent important targets for human gene therapy [14][15][16].…”

Section: Introductionmentioning

confidence: 99%

HIV-1-based defective lentiviral vectors efficiently transduce human monocytes-derived macrophages and suppress replication of wild-type HIV-1

et al. 2005

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Background-Human monocytes play an important role in mediating human immunodeficiency virus type 1 (HIV-1) infection of the central nervous system (CNS), and monocytes-derived macrophages (MDM) represent a major viral reservoir within the brain and other target organs. Current gene transduction of MDM is hindered by a limited efficiency. In this study we established a lentiviral vector-based technique for improved gene transfer into human MDM cultures in vitro and demonstrated significant protection of transduced MDM from super-infection with wild-type HIV-1.

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“…Based on the observation that monomyeloic cells are targets of the M-tropic strains of HIV, several groups have used lentiviral vectors successfully to introduce transgenes into macrophages (Burke et al, 2002). Two preliminary studies reported the feasibility of microglial transduction by lentiviruses (Wrzesinski et al, 2000;Mordelet et al, 2002), but no efficiencies or other details for microglial transduction were reported.…”

Section: Introductionmentioning

confidence: 99%

Lentiviral transduction of microglial cells

Balcaitis

Weinstein

et al. 2004

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Microglial cells are the resident immune cells of the central nervous system. Their function resembles that of tissue macrophages and, as such, they share many properties with both peripheral macrophages and monocytes. One striking similarity is the difficulty with which these cells can be genetically manipulated via transfection or transduction. We have sought to overcome this challenge and generate stably transduced microglial cell lines. Based on encouraging results from macrophages, we hypothesized that lentiviral vectors might provide a valuable tool in the transduction of microglial cells. Using a lentiviral-based vector system expressing enhanced green fluorescent protein (eGFP) under the control of the murine stem cell virus promoter (MSCV), we found that multiplicities of infection (MOIs) of 1, 10, and 100 transduce >70%, >88%, and >95% of the cells, respectively. From the pool of transduced cells, we established lines of N9 and BV-2 microglial cells with distinct fluorescence intensities. Using real time-polymerase chain reaction (PCR), we correlated the integrated eGFP copy numbers to eGFP fluorescence measured by flow cytometry. When mixed, up to three lines with different eGFP intensities could be separated by flow cytometry and fluorescence microscopy. Neither infection nor transgene expression influenced microglial activation as assessed by nitric oxide (NO) production, cytokine release, and surface antigen expression. Our findings that microglial cells are easily transduced by lentiviral based vectors will facilitate research depending on genetic manipulation and help generate transgenic cell lines. In addition, the availability of microglial cell lines with defined fluorescence properties could replace elaborate staining procedures for microglial identification in co-culture experiments.

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Brain engraftment of autologous macrophages transduced with a lentiviral flap vector: an approach to complement brain dysfunctions

Cited by 15 publications

References 42 publications

Selective gene expression in brain microglia mediated via adeno-associated virus type 2 and type 5 vectors

Selective gene expression in brain microglia mediated via adeno-associated virus type 2 and type 5 vectors

HIV-1-based defective lentiviral vectors efficiently transduce human monocytes-derived macrophages and suppress replication of wild-type HIV-1

Lentiviral transduction of microglial cells

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