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[通过免疫球蛋白重链聚合酶反应对B细胞淋巴增殖性疾病进行克隆性分析]

[Clonality analysis of B-cell lymphoproliferative disorders by means of immunoglobulin heavy chain polymerase reaction].

作者信息

László T, Kelényi G, Matolcsy A

机构信息

Pécsi Orvostudományi Egyetem, Pathologiai Intézet.

出版信息

Orv Hetil. 1996 Sep 8;137(36):1963-7.

PMID:8927349
Abstract

The majority of B-cell non-Hodgkin's lymphomas (NHL) exhibit a highly specific immunoglobulin heavy chain (IgH) gene rearrangement as a result of sequential assembly of their Ig variable (VH), diversity (D) and joining (JH) region segments. The analyses of Ig gene rearrangements in B cells may help to differentiate reactive lymphoproliferations from NHLs, and to identify of their B-cell origin. The aim of this study was to reveal the usefulness of polymerase chain reaction analysis of the Ig gene rearrangement in the diagnosis of B-cell NHLs, using native and formol-paraffin embedded samples. The authors analysed 67 biopsy samples of immunohistochemically characterized lymph nodes diagnosed at the Department of Pathology. University Medical School of Pécs, between 1993 and 1995, using IgH gene polymerase chain reaction. The 67 samples included 10 reactive lymphoproliferations, 47 B-cell, 5 T-cell NHLs and 5 Hodgkin's diseases. In 54 cases, fresh, unfixed, in 13 cases, formalin-fixed samples have been used. The polymerase chain reaction amplification of the Ig heavy chain third complementary determining region (CDR 3) was performed by IgVH specific sense and JH specific antisense primer pairs. The polymerase chain reaction products were evaluated by agarose gel electrophoresis containing ethidium bromide. Sixty-four % of fresh, unfixed and 54% of formol-paraffin fixed B-cell NHLs samples showed clonal Ig gene rearrangement. The applied polymerase chain reaction technique did not show clonal amplification in reactive lymphoproliferations, T-cell NHLs or Hodgkin's disease. The polymerase chain reaction amplification of the IgH gene can be a powerful tool in the diagnosis of monoclonal B-cell lymphoproliferative disorders.

摘要

大多数B细胞非霍奇金淋巴瘤(NHL)由于其免疫球蛋白重链(IgH)可变区(VH)、多样性区(D)和连接区(JH)片段的顺序组装而表现出高度特异性的IgH基因重排。分析B细胞中的Ig基因重排可能有助于区分反应性淋巴细胞增殖与NHL,并确定其B细胞起源。本研究的目的是通过使用新鲜和甲醛石蜡包埋样本,揭示Ig基因重排的聚合酶链反应分析在B细胞NHL诊断中的实用性。作者使用IgH基因聚合酶链反应分析了1993年至1995年间在佩奇大学医学院病理科诊断的67例经免疫组织化学特征化的淋巴结活检样本。这67个样本包括10例反应性淋巴细胞增殖、47例B细胞、5例T细胞NHL和5例霍奇金病。在54例中使用了新鲜、未固定的样本,在13例中使用了福尔马林固定的样本。通过IgVH特异性正向引物和JH特异性反向引物对进行免疫球蛋白重链第三互补决定区(CDR 3)的聚合酶链反应扩增。通过含溴化乙锭的琼脂糖凝胶电泳评估聚合酶链反应产物。64%的新鲜、未固定的B细胞NHL样本和54%的甲醛石蜡固定的B细胞NHL样本显示克隆性Ig基因重排。所应用的聚合酶链反应技术在反应性淋巴细胞增殖、T细胞NHL或霍奇金病中未显示克隆性扩增。IgH基因的聚合酶链反应扩增可以成为诊断单克隆B细胞淋巴细胞增殖性疾病的有力工具。

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