de Winde Charlotte M, Zuidscherwoude Malou, Vasaturo Angela, van der Schaaf Alie, Figdor Carl G, van Spriel Annemiek B
Department of Tumor Immunology, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, Geert Grooteplein-Zuid 26, 6525 GA, Nijmegen, The Netherlands.
Histochem Cell Biol. 2015 Aug;144(2):133-46. doi: 10.1007/s00418-015-1326-2. Epub 2015 May 8.
Multispectral imaging is a novel microscopy technique that combines imaging with spectroscopy to obtain both quantitative expression data and tissue distribution of different cellular markers. Tetraspanins CD37 and CD53 are four-transmembrane proteins involved in cellular and humoral immune responses. However, comprehensive immunohistochemical analyses of CD37 and CD53 in human lymphoid organs have not been performed so far. We investigated CD37 and CD53 protein expression on primary human immune cell subsets in blood and in primary and secondary lymphoid organs. Both tetraspanins were prominently expressed on antigen-presenting cells, with highest expression of CD37 on B lymphocytes. Analysis of subcellular distribution showed presence of both tetraspanins on the plasma membrane and on endosomes. In addition, CD53 was also present on lysosomes. Quantitative analysis of expression and localization of CD37 and CD53 on lymphocytes within lymphoid tissues by multispectral imaging revealed high expression of both tetraspanins on CD20(+) cells in B cell follicles in human spleen and appendix. CD3(+) T cells within splenic T cell zones expressed lower levels of CD37 and CD53 compared to T cells in the red pulp of human spleen. B cells in human bone marrow highly expressed CD37, whereas the expression of CD53 was low. In conclusion, we demonstrate differential expression of CD37 and CD53 on primary human immune cells, their subcellular localization and their quantitative distribution in human lymphoid organs. This study provides a solid basis for better insight into the function of tetraspanins in the human immune response.
多光谱成像是一种新型显微镜技术，它将成像与光谱学相结合，以获取不同细胞标志物的定量表达数据和组织分布。四跨膜蛋白CD37和CD53是参与细胞免疫和体液免疫反应的四跨膜蛋白。然而，迄今为止尚未对人淋巴器官中的CD37和CD53进行全面的免疫组织化学分析。我们研究了CD37和CD53蛋白在人血液、初级和次级淋巴器官中的原代免疫细胞亚群上的表达。这两种四跨膜蛋白在抗原呈递细胞上均有显著表达，其中CD37在B淋巴细胞上的表达最高。亚细胞分布分析显示，这两种四跨膜蛋白均存在于质膜和内体上。此外，CD53也存在于溶酶体上。通过多光谱成像对淋巴组织内淋巴细胞上CD37和CD53的表达和定位进行定量分析，结果显示，在人脾脏和阑尾的B细胞滤泡中的CD20(+)细胞上，这两种四跨膜蛋白均高表达。与人类脾脏红髓中的T细胞相比，脾脏T细胞区中的CD3(+)T细胞表达的CD37和CD53水平较低。人类骨髓中的B细胞高表达CD37，而CD53的表达较低。总之，我们证明了CD37和CD53在人原代免疫细胞上的差异表达、它们的亚细胞定位以及它们在人淋巴器官中的定量分布。这项研究为更好地了解四跨膜蛋白在人类免疫反应中的功能提供了坚实的基础。
