Stranneheim Henrik, Orre Lukas M, Lehtiö Janne, Flygare Jenny
Department of Gene Technology, AlbaNova University Center, Royal Institute of Technology, Stockholm, Sweden.
Proteome Sci. 2009 Nov 23;7:43. doi: 10.1186/1477-5956-7-43.
B-cell lymphomas are thought to reflect different stages of B-cell maturation. Based on cytogenetics and molecular markers, mantle cell lymphoma (MCL) is presumed to derive predominantly from naïve, pre-germinal centre (pre-GC) B lymphocytes. The aim of this study was to develop a method to investigate the similarity between MCL cells and different B-cell compartments on a protein expression level.
Subpopulations of B cells representing the germinal centre (GC), the pre-GC mantle zone and the post-GC marginal zone were isolated from tonsils using automated magnetic cell sorting (AutoMACS) of cells based on their expression of CD27 and IgD. Protein profiling of the B cell subsets, of cell lines representing different lymphomas and of primary MCL samples was performed using top-down proteomics profiling by surface-enhanced laser detection/ionization time-of-flight mass spectrometry (SELDI-TOF-MS).
Quantitative MS data of significant protein peaks (p-value < 0.05) separating the three B-cell subpopulations were generated. Together, hierarchical clustering and principal component analysis (PCA) showed that the primary MCL samples clustered together with the pre- and post-GC subpopulations. Both primary MCL cells and MCL cell lines were clearly separated from the B cells representing the GC compartment.
AutoMACS sorting generates sufficient purity to enable a comparison between protein profiles of B cell subpopulations and malignant B lymphocytes applying SELDI-TOF-MS. Further validation with an increased number of patient samples and identification of differentially expressed proteins would enable a search for possible treatment targets that are expressed during the early development of MCL.
B细胞淋巴瘤被认为反映了B细胞成熟的不同阶段。基于细胞遗传学和分子标志物，套细胞淋巴瘤（MCL）被推测主要来源于幼稚的生发中心前（pre-GC）B淋巴细胞。本研究的目的是开发一种方法，在蛋白质表达水平上研究MCL细胞与不同B细胞区室之间的相似性。
利用基于CD27和IgD表达的细胞自动磁性细胞分选（AutoMACS）技术，从扁桃体中分离出代表生发中心（GC）、生发中心前套区和生发中心后边缘区的B细胞亚群。通过表面增强激光检测/电离飞行时间质谱（SELDI-TOF-MS）进行自上而下的蛋白质组学分析，对B细胞亚群、代表不同淋巴瘤的细胞系和原发性MCL样本进行蛋白质谱分析。
生成了区分三个B细胞亚群的显著蛋白峰（p值<0.05）的定量质谱数据。层次聚类和主成分分析（PCA）共同显示，原发性MCL样本与生发中心前和生发中心后的亚群聚集在一起。原发性MCL细胞和MCL细胞系均与代表生发中心区室的B细胞明显分离。
AutoMACS分选产生了足够的纯度，能够使用SELDI-TOF-MS比较B细胞亚群和恶性B淋巴细胞的蛋白质谱。通过增加患者样本数量进行进一步验证并鉴定差异表达的蛋白质，将有助于寻找在MCL早期发育过程中表达的可能治疗靶点。
