Zandecki M, Bernardi F, Genevieve F, Laï J L, Preudhomme C, Flactif M, Cosson A, Bauters F, Facon T
Laboratoire d'Hématologie A, Hôpital Calmette, CHU, Lille, France.
Leukemia. 1997 Jul;11(7):1034-9.
The mononuclear cells in the peripheral blood are implicated in the myeloma process especially with the presence of peripheral blood plasma cells (PBPC) and clonal B lymphocytes found using phenotypic or gene rearrangement techniques. The purpose of this study was to look for aneuploidy in the two main B cell components of the peripheral blood: PBPC and CD20-positive B lymphocytes. Conventional cytogenetics (CC) or DNA content analysis and fluorescence in situ hybridization (FISH) with centromeric probes were performed on bone marrow plasma cells (BMPC) of 21 patients with multiple myeloma and peripheral blood cells were studied as follows: immunostaining to look for PBPC and to assess their number, image analysis cytometry for the determination of their DNA content, and FISH chromosomes analysis. FISH was performed using probes against the chromsomes that were lost or gained in BMPC and was coupled with immunostaining of the relevant light chain or CD20 antigen to study PBPC or B lymphocytes, respectively. Monotypic PBPC were found in 16 patients. Their DNA content was the same or nearly the same as for BMPC and they exhibited the same monosomies or trisomies as those found within their BM counterpart. By contrast, DNA content of mononuclear cells other than PBPC was within normal ranges, and in 13 of 15 patients CD20-positive B lymphocytes failed to show chromosomal changes by FISH analysis. In two patients however, a few CD20+ cells with lymphoid morphology exhibited chromosome changes, hypothesizing that a few cytogenetically abnormal B cells without plasmocytic morphology may circulate. From these data, we conclude that PBPC share the same genetic abnormalities as BMPC and thus belong to the malignant clone, whereas most peripheral blood B lymphocytes are unrelated to the tumor clone.
外周血中的单核细胞与骨髓瘤进程有关,尤其是在外周血浆细胞(PBPC)以及使用表型或基因重排技术发现的克隆性B淋巴细胞存在时。本研究的目的是在外周血的两种主要B细胞成分中寻找非整倍体:PBPC和CD20阳性B淋巴细胞。对21例多发性骨髓瘤患者的骨髓浆细胞(BMPC)进行了常规细胞遗传学(CC)或DNA含量分析以及着丝粒探针荧光原位杂交(FISH),外周血细胞研究如下:免疫染色以寻找PBPC并评估其数量,图像分析细胞术用于确定其DNA含量,以及FISH染色体分析。使用针对BMPC中缺失或增加的染色体的探针进行FISH,并分别与相关轻链或CD20抗原的免疫染色相结合,以研究PBPC或B淋巴细胞。在16例患者中发现了单型PBPC。它们的DNA含量与BMPC相同或几乎相同,并且表现出与骨髓对应物中发现的相同的单体或三体。相比之下,PBPC以外的单核细胞DNA含量在正常范围内,并且在15例患者中的13例中,CD20阳性B淋巴细胞通过FISH分析未显示染色体变化。然而,在两名患者中,一些具有淋巴细胞形态的CD20 +细胞表现出染色体变化,推测可能有一些无浆细胞形态的细胞遗传学异常B细胞在循环。根据这些数据,我们得出结论,PBPC与BMPC具有相同的遗传异常,因此属于恶性克隆,而大多数外周血B淋巴细胞与肿瘤克隆无关。