流式细胞免疫分型联合PCR检测抗原受体基因重排在淋巴增殖性疾病诊断与鉴别诊断中的应用

Application of flow cytometric immunophenotyping combined with polymerase chain reaction examination for antigen receptor gene rearrangement in the diagnosis of lymphoproliferative diseases

背景与目的:淋巴增殖性疾病是一组病理形态、免疫表型、临床特征高度异质性的疾病。本文旨在评价流式细胞免疫分型联合聚合酶链反应(PCR)检测抗原受体基因重排在淋巴增殖性疾病诊断中的应用价值。方法:分离79例拟诊淋巴增殖性疾病患者的新鲜病理标本(淋巴结、脾脏、胃活检组织和肝穿组织),流式细胞术分析细胞的免疫表型特征;同时采用BIOMED-2多重PCR分析组织的抗原受体基因重排状况及其克隆来源;并与病理诊断及免疫组织化学结果进行比对。结果:患者最终确诊为非霍奇金淋巴瘤(non-Hodgkin'slymphoma,NHL)37例(46.8%),霍奇金淋巴瘤(Hodgkin's lymphoma,HL)3例(3.8%),淋巴组织良性病变34例(43.0%),恶性肿瘤淋巴结转移4例(5.1%)。在19例B-NHL确诊患者中,流式免疫分型和抗原受体基因重排诊断符合率均为94.7%(18例):18例T-NHL确诊患者中,流式免疫分型和抗原受体基因重排诊断符合率分别为72.2%(13例)和50%(9例);在34例淋巴组织良性病变的患者中,流式细胞免疫分型虽未见单克隆幼稚细胞,但有7例(19.4%)抗原受体基因重排呈单克隆性;在3例HL确诊患者中,流式免疫分型未能检测出来,抗原受体基因重排均为多克隆性;4例恶性肿瘤转移患者中(3例转移浸润淋巴结,1例转移浸润肝脏),流式细胞免疫分型均检测到大量异常幼稚细胞,其中3例无特殊抗体标记,另外1例表达幼稚髓系标记;抗原受体基因重排均为多克隆性。结论:应用流式细胞免疫分型联合抗原受体基因重排分析,对于淋巴组织良、恶性增殖的鉴别和不同类型恶性淋巴瘤的诊断,具有很好的辅助价值。

Background and purpose：Lymphoproliferative disease is a disease of highly heterogeneous pathology,immunophenotype and clinical features.The study was aimed to estimate the diagnostic value of flow cytometric immunophenotyping combined with polymerase chain reaction for detection of antigen receptor gene rearrangements for lymphoproliferative diseases.Methods：Fresh samples（lymph nodes,spleen,gastric and hepatic tissue obtained by biopsy or operation）of 79 cases suspected as lymphoproliferative diseases were isolated and their immunophenotypes were examined by flow cytometry.Meanwhile,gene rearrangement and clonal origin of antigen receptors of these samples were analyzed by BIOMED-2 multiplex polymerase chain reaction.And the outcome was compared with pathological diagnosis and immunohistochemical results.Results：Thirty-seven of the 79 cases （46.8%）were confirmed as non-Hodgkin＇s lymphoma（NHL）,3 cases（3.8%）Hodgkin＇s lymphoma（HL）,34 cases （43.0%）benign lymphoid tissues,while 4 cases（5.1%）were finally diagnosed as metastatic malignant tumor.The accurate rates of flow cytometric immunophenotyping and antigen receptor gene rearrangement diagnosis in 19 cases confirmed as B-NHL were both 94.7%（18 cases）.Among 18 cases diagnosed as T-NHL,the conformity rate of the former was 72.2%（13 cases）and the latter was 50%（9 cases）.In the 34 cases proved to be benign lymphoid tissue, flow cytometric immunophenotyping showed no monoclonal naive cells.However,monoclonal rearrangements were found in 7 samples（16.7%）.Polyclonal rearrangements were found in the 3 cases of HL which failed to be confirmed by the flow cytometric immunophenotyping.And a large number of immature cells were detected by flow cytometric immunophenotyping in the 4 metastatic malignant tumor cases（3 with lymph node invasion,1 with liver metastasis）, of which no specific antibody was found in 3 cases,while immature myeloid marker was detected in 1 case.And polyclonal rearrangements were showed in all of these 4 samples.Conclusion：Combined use of flow cytometric immunophenotype and antigen receptor gene rearrangement analysis is good auxiliary method for differential diagnosis of lymphoproliferative diseases and identification of different types of malignant lymphoma,particularly in atypical cases.