免疫球蛋白重链基因重排检测对眼附属器淋巴增生性病变的诊断研究

The value of IgH gene rearrangement in the diagnosis of ocular adnexal lymphoproliferative lesions

目的探讨多聚酶链反应(PCR)检测免疫球蛋白重链(IgH)基因重排在眼附属器淋巴增生性病变良恶性鉴别中的应用价值。设计实验性研究。研究对象32例眼附属器淋巴增生性病变存档蜡块标本。方法应用PCR检测眼附属器淋巴增生性病变的IgH基因重排,结合常规HE染色和免疫组织化学染色结果进行分析。主要指标组织病理形态,免疫表型特征及基因重排形式。结果17例淋巴瘤中12例IgH基因呈单克隆性重排,阳性率为70.6%;10例反应性淋巴细胞增生中1例呈单克隆性重排,阳性率为10%。两者差异有统计学意义(P=0.004)。5例不典型淋巴细胞增生中,3例基因呈单克隆性重排,支持恶性淋巴瘤的诊断;2例呈多克隆性重排,支持良性反应性增生的诊断。结论依靠常规HE染色和免疫组织化学染色有时难以明确眼附属器淋巴增生性病变的良恶性,此时应用PCR检测病变的IgH基因重排,有助于鉴别其良恶性。(眼科,2008,17:33-36)

Objective To determine whether polymerase chain reaction （PCR） for the assay of B-cell IgH gene rearrangement in patients with ocular adnexal lymphoproliferative lesisons could be useful in the diagnosis of lymphoma, especially in differentiating a benign lesion from a malignant one. Design Experimental study, Participants Formalin-fixed and paraffin-embedded tissues of lymphoproliferative leasions of 32 patients. Main Outcome Measures Histopahthological and immunophenotypic features, the rearrangement of immunoglobulin heavy chain （IgH） gene. Methods In addition to clinical, pathological and immunohistochemical evaluations, 32 cases of orbital lymphoproliferative lesions were examined for IgH gene rearrangement by means of PCR to amplify the third frame work region （FR3） with formalin-fixed and paraffin-embedded tissues. Results The PCR using primer FR3A showed that 16 cases had discrete single products , which were within the molecular weight range of 100 to 120 bp , and were therefore interpreted as monoclonal. The positive rates of FR3 region of IgH gene monoclonal rearrangement in patients with malignant lymphoma, benign reactive lymphoid hyperplasia were70.6 % （12/ 17） , 10.0 % （1/ 10） respectively. The difference was statistically significant （Fisher＇s Exact Test, P〈0.05）. Three cases of atypical lymphoid hyperplasia showed monoclonal rearrangement of IgH gene. A faint smear pattern was demonstrated in the remaining 16 cases , representing polyclonal populations of lymphoid cells. Conclusion Molecular genetic analysis by PCR using the FR3A primer is helpful in the diagnosis of ocular adnexal lymphoproliferative lesions, especially those in which the diagnosis can not be made by routine histopathological findings and immunohistochemistry evaluation.