Utility of microdissection and polymerase chain reaction for the detection of immunoglobulin gene rearrangement and translocation in primary intraocular lymphoma
Utility of microdissection and polymerase chain reaction for the detection of immunoglobulin gene rearrangement and translocation in primary intraocular lymphoma
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DOI：
10.1016/s0161-6420(98)99036-4
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发表时间：
1998-09-01
期刊：
影响因子：
13.7
通讯作者：
Chan, CC
中科院分区：
文献类型：
--
作者：
Shen, DF;Zhuang, ZP;Chan, CC
目的：原发性眼内淋巴瘤是一种原发性中枢神经系统淋巴瘤，属于非霍奇金淋巴瘤。诊断通常通过鉴定玻璃体、眼、脑和脑脊液中的恶性大B淋巴细胞来进行;然而，这些细胞很少、易碎且难以识别。近年来，克隆性重链免疫球蛋白（IgH）基因重排和bcl-2基因易位已被报道在系统性B细胞淋巴瘤，并用于检测恶性细胞和作出诊断。作者调查了四名PCNSL患者的三只眼睛和脉络膜视网膜活检标本中的分子变化。设计：人体组织研究。材料：收集了五名PCNSL患者的眼部标本。干预：第一名患者进行了左眼诊断性眼球摘除术。第二例患者接受诊断性脉络膜视网膜活检。在第三个案例中，一对尸检的眼睛与反应性淋巴浆细胞浸润的患者与获得性免疫缺陷综合征（艾滋病）进行了研究。第4例为一例AIDS相关淋巴瘤患者的眼球摘除标本。主要观察指标：采用显微切割和聚合酶链反应（PCR）技术对常规、石蜡包埋、甲醛固定和冰冻组织切片中淋巴瘤细胞的bcl-2和IgH基因进行分析。上述4例淋巴瘤细胞均在V-H区的第三构架区发现IgH重排基因。结论：IgH基因重排可作为PCNSL的分子标记。显微切割允许从复杂的异质组织的组织切片中获得特定的、选择的、微小的细胞群并进行分析。IgH和bcl-2作为凋亡“存活”信号和原癌基因的易位可能参与PCNSL的发病。显微切割和PCR的结合是研究小病变和细胞群以及了解疾病机制的有力工具。
Objective: Primary intraocular lymphoma, a non-Hodgkin's lymphoma, is a primary central nervous system lymphoma (PCNSL). Diagnosis is usually made by identifying malignant, large B lymphocytes in the vitreous, eye, brain, and cerebral spinal fluid; however, these cells ave few, friable, and difficult to recognize. Recently, clonal heavy chain immunoglobulin (IgH) gene rearrangement and bcl-2 gene translocation have been reported in systemic B-cell lymphoma and are used for the detection of malignant cells and in making a diagnosis. The authors investigated the molecular changes in three eyes and a chorioretinal biopsy specimen of four patients with PCNSL.Design: Human tissue study.Materials: Five ocular specimens of PCNSL were collected.Intervention: The first patient had a diagnostic enucleation of the left eye. The second patient underwent diagnostic chorioretinal biopsy. In the third case, a pair of autopsied eyes with reactive lymphoplasmacytic infiltrates of a patient with acquired immune deficiency syndrome (AIDS) were studied. In the fourth case, an enucleated eye of a patient with AIDS-associated lymphoma was sampled.Main Outcome Measures: The bcl-2 and IgH genes of the lymphoma cells from routine, paraffin-embedded, formaldehyde-fixed, or frozen histologic tissue sections were analyzed using microdissection and polymerase chain reaction (PCR) technique.Results: Lymphoma cells obtained from the above four cases showed IgH rearrangement gene in the third framework of the V-H region. Bcl-2-associated translocation also was detected in three cases (cases 1, 2, and 4).Conclusion: Rearrangement of the IgH gene can serve as a molecular marker for PCNSL. Microdissection allows for procurement and analysis of specific, selected, minute cell populations that are obtained from histologic sections of the complex, heterogeneous tissue. Translocation of IgH and bcl-2, the apoptotic "survival" signal and proto-oncogene, could contribute to the pathogenesis of PCNSL. The combination of microdissection and PCR is a powerful tool for studies of small lesions and cell populations and for understanding disease mechanisms.