Multiphoton high-resolution 3D imaging of Langerhans cells and keratinocytes in the mouse skin model adopted for epidermal powdered immunization
Multiphoton high-resolution 3D imaging of Langerhans cells and keratinocytes in the mouse skin model adopted for epidermal powdered immunization
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DOI：
10.1038/sj.jid.5700290
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发表时间：
2006-07-01
影响因子：
6.5
通讯作者：
Tirlapur, Uday K.
中科院分区：
文献类型：
--
作者：
Mulholland, William J.;Arbuthnott, Edward A. H.;Tirlapur, Uday K.
朗格汉斯细胞（LC）可以用DNA包被的金微弹（“基因枪”）靶向以诱导有效的细胞和体液免疫应答。表皮内LC和角质形成细胞的相对体积分布可能影响基因枪免疫方案的功效。本研究采用近红外多光子激光扫描显微镜（NIR-MPLSM）对小鼠皮肤模型中LC和角质形成细胞的三维（3D）分布进行了定量分析。角质层（SC）和可行的表皮厚度与MPLSM测量发现与传统的组织学密切一致。LC位于垂直平面上，平均深度为14.9 μ m，在真皮-表皮边界上方小于3 μ m，并具有正态直方图分布。这可能对应于LC存在于基底上层（生发层）的事实。发现角质形成细胞的核体积比常驻LC的核体积（88.6 μ m）大约1.4倍（3）。重要的是，小鼠耳部皮肤中LC与角质形成细胞的比例（1：15）比人类乳房皮肤中报告的比例（1：53）高出三倍以上。因此，交叉呈递在临床基因枪应用中可能比在临床前小鼠研究中更重要。在使用小鼠模型的临床前试验中应考虑这些种间差异。
Langerhans cells (LCs) can be targeted with DNA-coated gold micro-projectiles ("Gene Gun'') to induce potent cellular and humoral immune responses. It is likely that the relative volumetric distribution of LCs and keratinocytes within the epidermis impacts on the efficacy of Gene Gun immunization protocols. This study quantified the three-dimensional (3D) distribution of LCs and keratinocytes in the mouse skin model with a near-infrared multiphoton laser-scanning microscope (NIR-MPLSM). Stratum corneum (SC) and viable epidermal thickness measured with MPLSM was found in close agreement with conventional histology. LCs were located in the vertical plane at a mean depth of 14.9 mu m, less than 3 mu m above the dermo-epidermal boundary and with a normal histogram distribution. This likely corresponds to the fact that LCs reside in the suprabasal layer (stratum germinativum). The nuclear volume of keratinocytes was found to be approximately 1.4 times larger than that of resident LCs (88.6 mu m(3)). Importantly, the ratio of LCs to keratinocytes in mouse ear skin (1:15) is more than three times higher than that reported for human breast skin (1:53). Accordingly, cross-presentation may be more significant in clinical Gene Gun applications than in pre-clinical mouse studies. These interspecies differences should be considered in pre-clinical trials using mouse models.