多参数流式细胞术在淋巴细胞白血病和非霍奇金淋巴瘤骨髓侵犯诊断的应用

背景与目的：骨髓形态学检查可诊断淋巴细胞白血病,结合淋巴结活检可诊断非霍奇金淋巴瘤(non—Hodgkin‘s lymphoma,NHL)侵犯骨髓,但形态学仅是初步的诊断,采用免疫分型而获得肿瘤细胞来源及发育阶段的资料是目前诊治淋巴系统恶性肿瘤所必需的。本研究探讨多参数流式细胞术(flow cytometry,FCM)在淋巴细胞白血病、NHL骨髓侵犯的诊断和免疫分型方面的应用价值。方法：取初治白血病患者骨髓标本11例和NHL患者骨髓侵犯的骨髓标本41例,以及2例分别表现为巨大纵隔肿块和腹部巨大肿块的患者因无法取得病理标本而取骨髓检测的骨髓标本2例。采用B细胞系列抗体、T细胞系列抗体、粒细胞系列抗体,按常规行FCM免疫表型检测,CD45结合两个系列单抗或阶段特异性单抗进行三色免疫荧光染色,CD45／SSC设门后可将骨髓细胞清晰地分出成熟细胞和幼稚细胞群,然后行FSC、SSC、McAb1-FITC、McAb2-PE、CD45-Cychrome五参数分析。结果：11例白血病患者骨髓形态学经FCM的免疫分型获得进一步确诊和分型。41例NHL骨髓侵犯标本免疫表型与其淋巴结病理免疫组化相符合的为80．5％(33／41),不相符的有19．5％(8／41),结合临床、病理、骨髓形态学诊断和骨髓FCM的结果,最终获得明确诊断。2例分别为巨大纵隔肿块和腹块的患者仅靠骨髓形态学和骨髓FCM确诊为T—NHL和B—NHL。结论：多参数FCM能进一步明确白血病和NHL骨髓侵犯的诊断,对急性淋巴细胞白血病和NHL的病例还可同时获得T或B细胞来源和细胞分化早期或后期的参数,有助于临床诊断、鉴别诊断和治疗方案的确定。

The value of multiparameter flow cytometry in diagnosis of lymphocytic leukemia and bone marrow involvement of non-Hodgkin's lymphoma

BACKGROUND & OBJECTIVE: Lymphocytic leukemia and bone marrow involvement of non-Hodgkin's lymphoma (NHL) can be diagnosed by bone marrow morphology or lymph node biopsy combined with bone marrow examination. The data of original and differentiation status of tumor can be analyzed by immunophenotype of bone marrow. These are necessary for diagnosis and treatment of lymphocytic malignancy. This study was designed to investigate the value of multiparameter flow cytometry in diagnosis of leukemia and bone marrow involvement of NHL. METHODS: The samples from 11 cases of untreated leukemia bone marrow and 41 cases of untreated NHL with bone marrow involvement and 2 cases of bone marrow whose biopsy could not be obtained due to huge mass in mediastinum and abdomen were detected by multiparameter flow cytometry using antibodies of T, B, Myeloid cell series. Three-color staining was done by CD45 combined with two cell series or special phase antibodies. Using CD45/SSC set gate to identify blast cells from mature cells. The samples were analyzed using five parameters [forward scatter (FSC), side scatter(SSC), McAb1-FITC, McAb2-PE, and CD45-cychrome]. RESULTS: Immunophenotype and diagnosis of 11 cases of leukemia were further confirmed by flow cytometry (FCM). Of 41 cases of NHL with bone marrow involvement, 33 cases (80.5%) lymph nodes immunophenotype by pathology diagnosis were consistent with bone marrow immunophenotype by FCM, 8 cases (19.5%) were inconsistent, but right diagnosis were made by combining with clinical presentation, pathology, bone marrow morphology and FCM. Another 2 cases with huge mediastinal mass and abdominal mass were diagnosed as T-NHL and B-NHL by bone marrow morphology and FCM without lymph node biopsy. CONCLUSION: Multiparameter flow cytometry of bone marrow can further ascertain the diagnosis of leukemia and NHL with bone marrow involvement. It also gives us data of cells lineage and differentiation status for leukemia and NHL. It is helpful for diagnosis, differential diagnosis, and treatment.