聚合酶链反应扩增IgH CDR-Ⅲ序列对眼部淋巴样增生病的诊断研究

目的从分子生物学角度探讨眼部淋巴样增生病的诊断、分类及肿瘤细胞的起源。方法应用聚合酶链反应（ｐｏｌｙｍｅｒａｓｅｃｈａｉｎｒｅａｃｔｉｏｎ,ＰＣＲ）对２９例（３１只眼）眼部淋巴样增生病的石蜡切片进行免疫球蛋白重链基因第三互补决定区（ＩｇＨＣＤＲ－Ⅲ）重排检测,并结合形态学及免疫表型进行分析检查。结果１７只眼恶性淋巴瘤、５只眼反应性增生均呈ＩｇＨ基因克隆性重排,电泳产物可见一明显的扩增窄带（１００～１２０ｂｐ）,其中２例双眼恶性淋巴瘤的两侧病变扩增带一致,１只眼恶性淋巴瘤、８只眼反应性增生均呈ＩｇＨ基因多克隆性重排。结论采用ＰＣＲ扩增ＩｇＨＣＤＲ－Ⅲ序列能确定此类疾病的克隆性质,并能发现免疫组化及形态学不能识别的小克隆Ｂ细胞病变,为诊断眼部淋巴样增生病提供了一个较为客观、敏感而又实用的手段。

A study on diagnosis of ocular adnexal lymphoid hyperplasia by polymerase chain reaction amplified IgH CDR-III sequence

OBJECTIVE: To study the diagnosis and classification of ocular adnexal lymphoid hyperplasia and investigate its cell origin by using some molecular biological techniques. METHODS: 29 cases (31 eyes) of ocular adnexal lymphoid hyperplasia were examined for immunoglobulin heavy-chain (IgH) gene rearrangements by means of polymerase chain reaction (PCR) to amplify the complementarity determining region III (CDR-III) with formalin-fixed and paraffin-embedded tissue combining with histopathologic examination and immunophenotypic analysis. RESULTS: 17 eyes of malignant lymphoma and 5 eyes of reactive lymphoid hyperplasia demonstrated clear single band of the rearranged gene for the IgH chain between 100 bp. to 120 bp. The bilateral ocular adnexal lymphoma occurring simultaneously in 4 eyes (two individuals) exhibited identical band. 1 eye of malignant lymphoma, 8 eyes of atypical lymphoid hyperplasias and 6 eyes of reactive lymphoid hyperplasia showed no band or a smear band. CONCLUSIONS: The results show that this method can determine the clonality of the lymphoid hyperplasias. It may be as well in the examination of small monoclonal B cell populations for which immunophenotypical analysis is difficult. The method is an objective, accurate and useful approach for the diagnosis of ocular adnexal lymphoid hyperplasias.