Identification of cell adhesion molecules in the human follicle-associated epithelium that improve nanoparticle uptake into the Peyer's patches
Identification of cell adhesion molecules in the human follicle-associated epithelium that improve nanoparticle uptake into the Peyer's patches
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DOI：
10.1124/jpet.106.107847
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发表时间：
2006-11-01
影响因子：
3.5
通讯作者：
Artursson, Per
中科院分区：
文献类型：
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作者：
Gullberg, Elisabet;Keita, Asa V.;Artursson, Per
这项研究的目的是确定可以作为覆盖在Peyer‘s补片上的人毛囊相关上皮(FAE)靶点的细胞黏附分子，并评估该上皮细胞对纳米颗粒的摄取水平。我们首先研究了小鼠M细胞标志物β(1)-整合素的表达，并利用来自肠上皮Caco-2细胞和Raji B细胞的人FAE模型，通过基因芯片鉴定了额外的潜在靶点。用免疫荧光法测定FAE模型和人回肠FAE组织中潜在靶点的蛋白表达。通过研究Arg-Gly-Asp(RGD)包被(整合素结合)、Arg-Gly-Glu(RGE)包覆(非整合素结合)和未包覆纳米颗粒在Ussing小室中安装的回肠样品中的传输，研究了整合素靶向。与Caco-2对照细胞或绒毛上皮细胞(VE)相比，β(1)-整合素和细胞黏附分子CD9在模型和人FAE中的表达更为丰富。未包覆的纳米颗粒在FAE或VE中都没有被吸收。以RGD为靶点的一般整合素显着改善了纳米颗粒在FAE中的传输，并在较低程度上改善了VE的传输。与RGE相比，RGD将跨FAE的传输提高了4倍。RGD和RGE包裹的纳米颗粒在VE中的传输没有差别。综上所述，β(1)-整合素和CD9被确定为人FAE的靶标。RGD和RGE介导的跨FAE的转运的差异表明，特定的整合素相互作用是改善纳米颗粒跨FAE摄取的主要机制，而电荷相互作用对VE摄取的改善有很大贡献。
The aim of this study was to identify cell adhesion molecules that could serve as targets of the human follicle-associated epithelium (FAE) overlying Peyer's patches and to assess nanoparticle uptake levels across this epithelium. We first studied the expression of the mouse M-cell marker beta(1)-integrin and used a model of human FAE derived from intestinal epithelial Caco-2 cells and Raji B-cells to identify additional potential targets by cDNA array. The protein expression of potential targets in the model FAE and in human ileal FAE tissues was quantified by immunofluorescence. Integrin targeting was studied by investigating the transport of Arg-Gly-Asp (RGD)-coated (integrin- binding), Arg-Gly-Glu (RGE)-coated (nonintegrin-binding), and uncoated nanoparticles across ileal specimens mounted in Ussing chambers. Both beta(1)-integrin and the cell adhesion molecule CD9 were more abundantly expressed in the model and human FAE compared with the Caco-2 control cells or villus epithelium (VE). Uncoated nanoparticles were not taken up across either FAE or VE. General integrin targeting with RGD improved the nanoparticle transport dramatically across the FAE and to a lower extent across the VE. Compared with RGE, RGD improved transport 4-fold across the FAE. There was no difference in the transport of RGD- and RGE-coated nanoparticles across the VE. In conclusion, beta(1)-integrin and CD9 were identified as targets in human FAE. The difference in RGD- and RGE-mediated transport across the FAE, but not the VE, suggests that a specific integrin interaction was the dominating mechanism for improved nanoparticle uptake across the FAE., whereas charge interaction contributed substantially to the improved VE uptake.