Transduced monocyte/macrophages targeted to murine skin by UV light.
Transduced monocyte/macrophages targeted to murine skin by UV light.
复制标题
通过紫外线将转导的单核细胞/巨噬细胞靶向小鼠皮肤。
DOI：
10.1111/j.0906-6705.2005.00394.x
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发表时间：
2006
期刊：
影响因子：
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通讯作者：
Gilliam,AnitaC
中科院分区：
文献类型：
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作者：
Zhang,AlexandraY;Wu,Caiyun;Zhou,Lixin;Ismail,SaharA;Tao,Jianming;McCormick,LauraL;Cooper,KevinD;Gilliam,AnitaC
Abstract:We have selectively targeted monocyte/macrophages overexpressing an immunomodulatory molecule, latency‐associated peptide (LAP), a naturally occurring antagonist for transforming growth factor‐β1, to murine skin utilizing UV light to produce a cutaneous influx of transduced monocyte/macrophages. Bone marrow (BM) cells from BALB/c mice were transducedin vitrowith a retroviral construct containing green fluorescent protein (GFP) for detection and human LAP (hLAP) as a test molecule. The transduced BM cells were then culturedin vitrowith granulocyte‐macrophage colony‐stimulating factor (GM‐CSF) to produce differentiation to monocyte/macrophages. More than 80% of transduced BM cells were CD11b‐positive and MOMA‐positive by fluorescence‐activated cell‐sorter analysis and secreted LAP by ELISA after 10 days of culture in granulocyte‐macrophage colony‐stimulating factor (GM‐CSF). Transduced monocyte/macrophages containing either GFP or hLAP‐GFP were then injected intravenously into BALB/c mice. One‐half of recipients in each group were exposed to UVB (72 mJ) to induce monocyte/macrophage infiltration into skin. Recipients were sacrificed 60 h after UV irradiation. We found transduced cutaneous macrophages expressing GFP by examining with fluorescence microscopy frozen skin sections of recipient mice immunostained with antibodies to GFP and to macrophage marker F4/80. We identified hLAP sequences by polymerase chain reaction (PCR) of total DNA in recipient blood and UV‐irradiated skin but not in unirradiated skin. LAP sequences were also detected at much lower levels in other organs (lung, spleen, and liver) by PCR. Therefore, we have shown that genetically altered monocytic cells can be injected intravenously and targeted to mouse skin by UV exposure. This macrophage‐based gene‐transfer method may be a potentially useful immunotherapeutic approach for delivering monocyte/macrophage‐derived products to skin.
