通过流式细胞术检测B细胞淋巴增殖性疾病中的细胞周期蛋白D1

Detection of cyclin D1 in B cell lymphoproliferative disorders by flow cytometry.

作者信息

Jain P, Giustolisi G M, Atkinson S, Elnenaei M O, Morilla R, Owusu-Ankomah K, Rafiq-Mohammed F, Matutes E, Wotherspoon A, Catovsky D

机构信息

Department of Academic Haematology and Cytogenetics, The Royal Marsden NHS Trust, Fulham Road, London SW3 6JJ, UK.

出版信息

J Clin Pathol. 2002 Dec;55(12):940-5. doi: 10.1136/jcp.55.12.940.

DOI:10.1136/jcp.55.12.940

PMID:12461064

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1769833/

Abstract

AIMS

To describe and revise a flow cytometric assay for evaluating cyclin D1 overexpression in B cell lymphoproliferative disorders (B-LPDs).

METHODS

Cyclin D1 expression was evaluated in 11 healthy controls and 51 patients with B-LPD by flow cytometry using the 5D4 monoclonal antibody. In 25 cases, experiments were repeated up to four times with mononuclear cells (MNC) fixed in ethanol for 1-120 days to evaluate the consistency of cyclin D1 expression. Flow cytometry results were compared with fluorescence in situ hybridisation (FISH) for the t(11;14) translocation in 19 patients and with immunohistochemistry (IHC) using the DCS-6 monoclonal antibody in nine patients.

RESULTS

A mean fluorescence intensity ratio (MFIR) of 4.8 was defined as the cut off point for positivity based on cyclin D1 expression in healthy controls (mean + 3 SD). Ten patients overexpressed cyclin D1 by flow cytometry. These included five of eight patients with mantle cell lymphoma, four of 19 with chronic lymphocytic leukaemia, and one with follicular lymphoma. MFIR in the repeat experiments differed less than 25% in 20 of 25 patients and in no cases did it cross the cut off point. There was a good correlation between cyclin D1 expression by flow cytometry and FISH for t(11;14) in 15 of 19 patients and six of nine had concordant results with flow cytometry, FISH, and IHC.

CONCLUSION

Cyclin D1 expression remains fairly stable once MNC are fixed in ethanol and the flow cytometric assay can be used for the routine screening of B-LPD. Further comparisons between flow cytometry, IHC, and FISH may be needed to ascertain the diagnostic value of the flow cytometric assay.

摘要

目的

描述并修订一种用于评估B细胞淋巴增殖性疾病（B-LPDs）中细胞周期蛋白D1过表达的流式细胞术检测方法。

方法

使用5D4单克隆抗体，通过流式细胞术对11名健康对照者和51例B-LPD患者的细胞周期蛋白D1表达进行评估。在25例患者中，用乙醇固定单核细胞（MNC）1至120天，重复实验多达4次，以评估细胞周期蛋白D1表达的一致性。将流式细胞术结果与19例患者的t(11;14)易位荧光原位杂交（FISH）结果以及9例患者使用DCS-6单克隆抗体的免疫组织化学（IHC）结果进行比较。

结果

基于健康对照者中细胞周期蛋白D1的表达情况（均值 + 3标准差），将平均荧光强度比（MFIR）4.8定义为阳性的截断点。通过流式细胞术检测，有10例患者细胞周期蛋白D1过表达。其中包括8例套细胞淋巴瘤患者中的5例、19例慢性淋巴细胞白血病患者中的4例以及1例滤泡性淋巴瘤患者。在25例患者中的20例，重复实验中的MFIR差异小于25%，且在任何情况下均未超过截断点。在19例患者中的15例，流式细胞术检测的细胞周期蛋白D1表达与t(11;14)的FISH结果之间具有良好的相关性，9例患者中的6例，流式细胞术、FISH和IHC结果一致。