Centre for Neurosciences and Cell Biology, Faculty of Pharmacy, University of Coimbra, Portugal.
Am J Pathol. 2012 Nov;181(5):1749-61. doi: 10.1016/j.ajpath.2012.07.033. Epub 2012 Sep 13.
Meningiomas are primary tumors of the central nervous system composed of both neoplastic and other infiltrating cells. We determined the cellular composition of 51 meningioma samples by multiparameter flow cytometric (MFC) immunophenotyping and investigated the potential relationship between mRNA and protein expression levels of neoplastic cells. For immunophenotypic, morphologic, and cytogenetic characterization of individual cell populations, a large panel of markers was used together with phagocytic/endocytic functional assays and MFC sorting. Overall, our results revealed coexistence of CD45(-) neoplastic cells and CD45(+) immune infiltrating cells in all meningiomas. Infiltrating cells included tissue macrophages, with an HLA-DR(+)CD14(+)CD45(+)CD68(+)CD16(-/+)CD33(-/+) phenotype and high phagocytic/endocytic activity, and a small proportion of cytotoxic lymphocytes (mostly T CD8(+) and natural killer cells). Tumor cells expressed multiple cell adhesion proteins, tetraspanins, HLA-I/HLA-DR molecules, complement regulatory proteins, cell surface ectoenzymes, and growth factor receptors. Noteworthy, the relationship between mRNA and protein levels was variable, depending on the proteins evaluated and the level of infiltration by immune cells. In summary, our results indicate that MFC immunophenotyping provides a reliable tool for the characterization of the patterns of protein expression of different cell populations coexisting in meningioma samples, with a more accurate measure of gene expression profiles of tumor cells at the functional/protein level than conventional mRNA microarray, independently of the degree of infiltration of the tumor by immune cells.
脑膜瘤是由肿瘤细胞和其他浸润细胞组成的中枢神经系统原发性肿瘤。我们通过多参数流式细胞术(MFC)免疫表型分析确定了 51 例脑膜瘤样本的细胞组成,并研究了肿瘤细胞 mRNA 和蛋白表达水平之间的潜在关系。为了对单个细胞群体进行免疫表型、形态学和细胞遗传学特征分析,我们使用了大量的标志物,并结合吞噬/内吞功能测定和 MFC 分选。总的来说,我们的结果表明所有脑膜瘤中均存在 CD45(-)肿瘤细胞和 CD45(+)免疫浸润细胞。浸润细胞包括组织巨噬细胞,其表型为 HLA-DR(+)CD14(+)CD45(+)CD68(+)CD16(-/+)CD33(-/+),具有高吞噬/内吞活性,以及一小部分细胞毒性淋巴细胞(主要为 T CD8(+)和自然杀伤细胞)。肿瘤细胞表达多种细胞黏附蛋白、四跨膜蛋白、HLA-I/HLA-DR 分子、补体调节蛋白、细胞表面外切酶和生长因子受体。值得注意的是,mRNA 和蛋白水平之间的关系因所评估的蛋白和免疫细胞浸润程度而异。总之,我们的结果表明,MFC 免疫表型分析为研究共存于脑膜瘤样本中的不同细胞群体的蛋白表达模式提供了可靠的工具,与传统的 mRNA 微阵列相比,它可以更准确地测量肿瘤细胞在功能/蛋白水平上的基因表达谱,而不受肿瘤细胞浸润程度的影响。