Zucchetto Antonella, Sonego Paolo, Degan Massimo, Bomben Riccardo, Dal Bo Michele, Russo Stefania, Attadia Vincenza, Rupolo Maurizio, Buccisano Francesco, Steffan Agostino, Del Poeta Giovanni, Pucillo Carlo, Colombatti Alfonso, Campanini Renato, Gattei Valter
Clinical and Experimental Hematology Research Unit, Centro di Riferimento Oncologico, I.R.C.C.S., Via Pedemontana Occidentale, 12, Aviano (PN), Italy.
J Immunol Methods. 2005 Oct 20;305(1):20-32. doi: 10.1016/j.jim.2005.07.004. Epub 2005 Aug 8.
Studies of gene expression profiling (GEP) have been successfully used for the identification of molecules to be employed as potential prognosticators. With the aim of identifying the immunophenotypic profile of B-CLL subsets with different prognoses, we investigated by flow cytometry the expression of 36 surface antigens in 117 cases, 113 with survival data. In analogy with GEP, results were analyzed by applying unsupervised hierarchical algorithms (surface-antigen expression profiling, SEP). Distinct immunophenotypic groups (A, B1, B2 and C) were identified, group C (57/117) with longer survivals, as compared to groups A (23/117), B1 (16/117) and B2 (21/117). The immunophenotypic signatures of these groups were characterized by the coordinated and differential over-expression of: i) CD62L, CD54 and CD49c (group C); ii) CD38 and CD49d (group A); iii) none of the above markers (group B1 and B2). Other molecules were either not expressed, widely expressed by all samples, or were variably expressed within the observed B-CLL subgroups, although without a clearly distinguishable pattern. By employing an identical approach for investigating the reactivity of B-cell panel monoclonal antibodies (B-mAbs) in B-CLLs (29 cases) and in 19 B and non-B leukemia/lymphoma cell lines, we found mAbs (B012, B001, B006, B018, B019, B020, B017) mainly unreactive in all the samples, mAbs (B002, B010, B013, B014, B015) strongly reactive in B-CLLs and B-cell lines but not in non-B-cell lines, and mAbs recognizing antigens variably expressed in cell lines and B-CLLs. A hierarchical clustering focused on B-CLLs alone, combining reactivity values for B-mAbs with the expression of CD62L and CD38, these latter antigens identified as leader markers of B-CLL subsets with different prognosis, demonstrated a correlation between CD62L expression and the reactivity of B007, B003, B011 and B005 mAbs. These mAbs may represent potentially novel markers with prognostic relevance in B-CLLs.
基因表达谱（GEP）研究已成功用于鉴定可作为潜在预后指标的分子。为了确定具有不同预后的B细胞慢性淋巴细胞白血病（B-CLL）亚群的免疫表型谱，我们通过流式细胞术研究了117例患者中36种表面抗原的表达情况，其中113例有生存数据。与GEP类似，通过应用无监督分层算法（表面抗原表达谱，SEP）对结果进行分析。确定了不同的免疫表型组（A、B1、B2和C），与A组（23/117）、B1组（16/117）和B2组（21/117）相比，C组（57/117）的生存期更长。这些组的免疫表型特征表现为以下分子的协同和差异过度表达：i）CD62L、CD54和CD49c（C组）；ii）CD38和CD49d（A组）；iii）上述标记均无（B1组和B2组）。其他分子要么不表达，要么在所有样本中广泛表达，要么在观察到的B-CLL亚组中可变表达，尽管没有明显可区分的模式。通过采用相同的方法研究B细胞面板单克隆抗体（B-mAbs）在B-CLL（29例）以及19种B和非B白血病/淋巴瘤细胞系中的反应性，我们发现mAbs（B012、B001、B006、B018、B019、B020、B017）在所有样本中主要无反应，mAbs（B002、B010、B013、B014、B015）在B-CLL和B细胞系中强烈反应，但在非B细胞系中无反应，还有一些mAbs识别在细胞系和B-CLL中可变表达的抗原。仅针对B-CLL进行的分层聚类，将B-mAbs的反应性值与CD62L和CD38的表达相结合，后两种抗原被确定为具有不同预后的B-CLL亚群的主导标记，结果表明CD62L表达与B007、B003、B011和B005 mAbs的反应性之间存在相关性。这些mAbs可能代表B-CLL中具有预后相关性的潜在新标记。
