Tyrer Peter, Ruth Foxwell A, Kyd Jennelle, Harvey Matthew, Sizer Phillip, Cripps Allan
Gadi Research Centre, Division of Science and Design, University of Canberra, ACT 2601, Canberra, Australia.
Biochem Biophys Res Commun. 2002 Dec 6;299(3):377-83. doi: 10.1016/s0006-291x(02)02631-1.
This study has evaluated an in vitro model of the follicle-associated epithelia that overlie Peyer's patches of the small intestine. The model shares many phenotypic characteristics of M cells in vivo. Co-cultures of the human adenocarcinoma cell line Caco-2 and freshly isolated Peyer's patch cells were established. Fluorescence microscopy and quantitative image analysis were used to validate the model against known markers of M-cell phenotype. Apical expression of alkaline phosphatase was down-regulated in co-cultures and villin was re-distributed from the apical membrane to the cytoplasm. alpha5beta1 integrin was found on the apical surfaces of the monolayers and B and T lymphocytes integrated into the monolayers. Particle transport was temperature-dependent in co-cultures, indicating that a transcytotic route was responsible. This model provides opportunities to study factors that influence M-cell development, assess putative Peyer's patch targeting in oral vaccine technologies, and study intestinal uptake in vitro.
本研究评估了覆盖小肠派尔集合淋巴结的滤泡相关上皮的体外模型。该模型具有体内M细胞的许多表型特征。建立了人腺癌细胞系Caco-2与新鲜分离的派尔集合淋巴结细胞的共培养体系。利用荧光显微镜和定量图像分析，根据已知的M细胞表型标志物对该模型进行验证。在共培养体系中，碱性磷酸酶的顶端表达下调，绒毛蛋白从顶端膜重新分布到细胞质中。在单层细胞的顶端表面发现了α5β1整合素，并且B和T淋巴细胞整合到了单层细胞中。在共培养体系中，颗粒转运是温度依赖性的，这表明存在一条转胞吞途径。该模型为研究影响M细胞发育的因素、评估口服疫苗技术中假定的派尔集合淋巴结靶向性以及体外研究肠道摄取提供了机会。
