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Lentiviral vectors containing mouse Csf1r control elements direct macrophage-restricted expression in multiple species of birds and mammals.

Pridans C, Lillico S, Whitelaw B, Hume DA - Mol Ther Methods Clin Dev (2014)

Bottom Line: We have created a lentiviral construct containing mouse FIRE and promoter.The lentivirus is capable of directing macrophage-restricted reporter gene expression in mouse, rat, human, pig, cow, sheep, and even chicken.Rat bone marrow cells transduced with the lentivirus were capable of differentiating into macrophages expressing the reporter gene in vitro.

View Article: PubMed Central - PubMed

Affiliation: The Roslin Institute, Royal (Dick) School of Veterinary Studies, University of Edinburgh, Scotland , UK.

ABSTRACT
The development of macrophages requires signaling through the lineage-restricted receptor Csf1r. Macrophage-restricted expression of transgenic reporters based upon Csf1r requires the highly conserved Fms-intronic regulatory element (FIRE). We have created a lentiviral construct containing mouse FIRE and promoter. The lentivirus is capable of directing macrophage-restricted reporter gene expression in mouse, rat, human, pig, cow, sheep, and even chicken. Rat bone marrow cells transduced with the lentivirus were capable of differentiating into macrophages expressing the reporter gene in vitro. Macrophage-restricted expression may be desirable for immunization or immune response modulation, and for gene therapy for lysosomal storage diseases and some immunodeficiencies. The small size of the Csf1r transcription control elements will allow the insertion of large "cargo" for applications in gene therapy and vaccine delivery.

No MeSH data available.


Related in: MedlinePlus

Murine Csf1r:EGFP-FIRE transduced primary macrophages from multiple species express EGFP. The following cells were differentiated into macrophages with rhCSF1 and then transduced with the lentivirus: (a) pig bone marrow–derived macrophages (BMDM), (b) cow monocyte-derived macrophages (MDM), (c) human MDM. A chicken macrophage cell line, HD11 (d) and sheep alveolar macrophages (e) were also transduced. These cells were not selected in blasticidin. Bar = 20 µm. Transductions were performed at least twice for each cell type.
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fig4: Murine Csf1r:EGFP-FIRE transduced primary macrophages from multiple species express EGFP. The following cells were differentiated into macrophages with rhCSF1 and then transduced with the lentivirus: (a) pig bone marrow–derived macrophages (BMDM), (b) cow monocyte-derived macrophages (MDM), (c) human MDM. A chicken macrophage cell line, HD11 (d) and sheep alveolar macrophages (e) were also transduced. These cells were not selected in blasticidin. Bar = 20 µm. Transductions were performed at least twice for each cell type.

Mentions: The FIRE element is highly conserved from birds to humans. We therefore examined whether murine Csf1r:EGFP-FIRE could direct the expression of EGFP in macrophages of other species. Figure 4 clearly shows EGFP expression in BM-derived macrophages from pig (Figure 4a), monocyte-derived macrophages from cow (Figure 4b) and human (Figure 4c) as well as a chicken macrophage cell line (HD11, Figure 4d), and sheep alveolar macrophages (Figure 4e).


Lentiviral vectors containing mouse Csf1r control elements direct macrophage-restricted expression in multiple species of birds and mammals.

Pridans C, Lillico S, Whitelaw B, Hume DA - Mol Ther Methods Clin Dev (2014)

Murine Csf1r:EGFP-FIRE transduced primary macrophages from multiple species express EGFP. The following cells were differentiated into macrophages with rhCSF1 and then transduced with the lentivirus: (a) pig bone marrow–derived macrophages (BMDM), (b) cow monocyte-derived macrophages (MDM), (c) human MDM. A chicken macrophage cell line, HD11 (d) and sheep alveolar macrophages (e) were also transduced. These cells were not selected in blasticidin. Bar = 20 µm. Transductions were performed at least twice for each cell type.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4362345&req=5

fig4: Murine Csf1r:EGFP-FIRE transduced primary macrophages from multiple species express EGFP. The following cells were differentiated into macrophages with rhCSF1 and then transduced with the lentivirus: (a) pig bone marrow–derived macrophages (BMDM), (b) cow monocyte-derived macrophages (MDM), (c) human MDM. A chicken macrophage cell line, HD11 (d) and sheep alveolar macrophages (e) were also transduced. These cells were not selected in blasticidin. Bar = 20 µm. Transductions were performed at least twice for each cell type.
Mentions: The FIRE element is highly conserved from birds to humans. We therefore examined whether murine Csf1r:EGFP-FIRE could direct the expression of EGFP in macrophages of other species. Figure 4 clearly shows EGFP expression in BM-derived macrophages from pig (Figure 4a), monocyte-derived macrophages from cow (Figure 4b) and human (Figure 4c) as well as a chicken macrophage cell line (HD11, Figure 4d), and sheep alveolar macrophages (Figure 4e).

Bottom Line: We have created a lentiviral construct containing mouse FIRE and promoter.The lentivirus is capable of directing macrophage-restricted reporter gene expression in mouse, rat, human, pig, cow, sheep, and even chicken.Rat bone marrow cells transduced with the lentivirus were capable of differentiating into macrophages expressing the reporter gene in vitro.

View Article: PubMed Central - PubMed

Affiliation: The Roslin Institute, Royal (Dick) School of Veterinary Studies, University of Edinburgh, Scotland , UK.

ABSTRACT
The development of macrophages requires signaling through the lineage-restricted receptor Csf1r. Macrophage-restricted expression of transgenic reporters based upon Csf1r requires the highly conserved Fms-intronic regulatory element (FIRE). We have created a lentiviral construct containing mouse FIRE and promoter. The lentivirus is capable of directing macrophage-restricted reporter gene expression in mouse, rat, human, pig, cow, sheep, and even chicken. Rat bone marrow cells transduced with the lentivirus were capable of differentiating into macrophages expressing the reporter gene in vitro. Macrophage-restricted expression may be desirable for immunization or immune response modulation, and for gene therapy for lysosomal storage diseases and some immunodeficiencies. The small size of the Csf1r transcription control elements will allow the insertion of large "cargo" for applications in gene therapy and vaccine delivery.

No MeSH data available.


Related in: MedlinePlus