Discordant Bone Marrow Involvement in Diffuse Large B-Cell Lymphoma: Comparative Molecular Analysis Reveals a Heterogeneous Group of Disorders
Discordant Bone Marrow Involvement in Diffuse Large B-Cell Lymphoma: Comparative Molecular Analysis Reveals a Heterogeneous Group of Disorders
复制标题
弥漫性大 B 细胞淋巴瘤中骨髓受累不一致：比较分子分析揭示了一组异质性疾病
作者：
M. Kremer;Martin S Spitzer;S. Mandl–Weber;K. Stecker;B. Schmidt;H. Höfler;L. Quintanilla‐Martinez;F. Fend
诊断为大细胞非霍奇金淋巴瘤（NHL）的患者中不一致的骨髓（BM）受累的特征是骨髓浸润主要由小淋巴样细胞组成，细胞学上与低度NHL一致。虽然这种现象在形态学上得到了很好的描述，但缺乏关于这两种病变关系的分子数据。本研究的目的是通过使用免疫球蛋白重链基因（IgH），以及bcl-2重排，作为分子标记，探讨克隆关系的不协调淋巴瘤的表现。IgH重排通过PCR用针对框架区3或2（FR 3和FR 2）的共有引物扩增，随后在选定的情况下进行自动化片段长度分析和测序。bcl-2基因的重排被鉴定为针对主要断裂点区域的引物。通过激光捕获显微切割分离小BM浸润。此外，使用抗CD 3、CD 10、CD 20、bcl-2、bcl-6、p53和Ki 67抗原的抗体在石蜡切片上进行免疫组织化学。对21例弥漫性大B细胞淋巴瘤（DLBCL）石蜡包埋组织进行分析。免疫组化染色后，5例被排除，无论是一致的BM浸润的大细胞淋巴瘤与丰富的反应性T细胞（2例）或良性，反应性淋巴浸润（3例），证实了多克隆模式的IgH分析。在其余16例中，8例通过存在相同的克隆性IgH重排或bcl-2重排证实了共同的克隆起源。在4例中，识别不同的IgH或bcl-2重排提供了证据，存在两个克隆无关的肿瘤。其余4例病例因技术原因无法评价。形态学、表型和分子生物学结果与大多数病例的生殖中心起源的淋巴瘤相符。然而，在4例病例中，BM浸润的流式细胞术分析显示B细胞慢性淋巴细胞白血病表型。其中两例与DLBCL克隆相关，因此代表了Richter转化。总之，DLBCL中不一致的BM浸润代表了一组异质性疾病，包括具有克隆相关、临床隐匿性小细胞成分的病例，以及在不同部位同时出现两种克隆不同、不相关的B细胞肿瘤的病例。
Discordant bone marrow (BM) involvement in patients with a diagnosis of large-cell non-Hodgkin’s lymphoma (NHL) is characterized by marrow infiltrates predominantly composed of small lymphoid cell, cytologically compatible with low-grade NHL. Although this phenomenon is well described morphologically, molecular data concerning the relationship of the two lesions are lacking. The aim of the study was to investigate the clonal relationship of discordant lymphoma manifestations by using immunoglobulin heavy chain gene (IgH), as well as bcl-2 rearrangements, as molecular markers. IgH rearrangements were amplified by PCR with consensus primers directed against framework regions 3 or 2 (FR3 and FR2), followed by automated fragment length analysis and sequencing in selected cases. Rearrangements of the bcl-2 gene were identified with primers against the major breakpoint region. Small BM infiltrates were isolated by laser capture microdissection. In addition, immunohistochemistry was performed on paraffin sections using antibodies against CD3, CD10, CD20, bcl-2, bcl-6, p53, and the Ki67 antigen. Paraffin-embedded tissues of 21 cases diagnosed as diffuse large B-cell lymphoma (DLBCL) with discordant BM involvement and no previous history of low-grade B-cell NHL were analyzed. After review of immunohistochemical stains, 5 cases were excluded either as concordant BM infiltrates by large-cell lymphoma with abundant reactive T-cells (2 cases) or as benign, reactive lymphoid infiltrates (3 cases), as confirmed by a polyclonal pattern in the IgH analysis. Of the remaining 16 cases, a common clonal origin was confirmed in 8 cases by the presence of an identical clonal IgH rearrangement or bcl-2 rearrangement. In 4 cases, identification of distinct IgH or bcl-2 rearrangements gave evidence for the presence of two clonally unrelated neoplasms. The remaining 4 cases were not evaluable for technical reasons. Morphological, phenotypical, and molecular findings were compatible with a lymphoma of germinal center origin in the majority of cases. However, in 4 cases, flow cytometric analysis of the BM infiltrates revealed a B-cell chronic lymphocytic leukemia phenotype. Two of these cases were clonally related to the DLBCL and thus represented Richter’s transformation. In summary, discordant BM infiltrates in DLBCL represent a heterogeneous group of disorders, encompassing cases with a clonally related, clinically occult small-cell component, as well as cases with two clonally distinct, unrelated B-cell neoplasms presenting synchronously at different locations.
影响因子：
158.5
作者：
WEISS, LM;WARNKE, RA;CLEARY, ML
通讯作者：
CLEARY, ML