流式细胞术对非霍奇金氏淋巴瘤的诊断价值评价

目的:探讨流式细胞术对非霍奇金氏淋巴瘤(NHL)的诊断价值。方法:收集郑州市中医院2009年1月至2014年8月收治的经病理组织学、免疫组织化学确诊的NHL患者20例临床资料。标本采集分为淋巴结细针穿刺13例,骨髓穿刺标本2例,外周血标本2例,淋巴结活检标本3例。结果:经FCM免疫分型、细胞涂片观察后,共有B-SLL6例,B-NHL 1例,FL 1例,DLBCL 5例,T-NHL 2例,T-LBL 3例,NK/T-NHL 1例,PTun 1例。结论:流式细胞术对NHL有一定的诊断价值,能够辅助临床诊断。     

流式细胞术在淋巴瘤免疫表型分析中的作用

目的探讨流式细胞术免疫表型分析在淋巴瘤诊断中的应用。方法采用流式细胞术对92例淋巴瘤患者进行免疫表型分析。结果在92例受检者中,B细胞NHL67例,T细胞NHL25例。B细胞NHL标记抗原出现频率较高的为CD19、CD22;T细胞NHL标记抗原出现频率最高的为CD7。结论流式细胞术在淋巴瘤的诊断分型方面有重要意义。更多还原     

流式细胞免疫表型分析在恶性淋巴瘤诊断中的应用

目的探讨多参数流式细胞免疫表型（FCI）分析在淋巴瘤诊断及鉴别诊断中的应用价值；诠释恶性淋巴瘤亚群免疫表型和分类，以进一步确定该技术在常规临床病理诊断中应用的可行性。方法收集2004年11月至2006年8月临床怀疑恶性淋巴瘤病例的新鲜组织标本74例，运用FCI技术检测分析每例标本的免疫表型，总结恶性淋巴瘤的FCI特征，并对比FCI结果与组织学诊断之间的符合率。结果74例FCI结果显示，61例（82．4％）与组织学结果相符，其中B细胞非霍奇金淋巴瘤（B-NHL）29例，T细胞非霍奇金淋巴瘤（T-NHL）6例，胸腺瘤6例，癌7例，淋巴结良性病变13例，与组织病理学结果相符率分别为93．5％、100％、100％、100％和81．3％。结论多参数FCI分析在NHL的诊断、分类及亚型判定中具有重要价值，是恶性淋巴瘤诊断和鉴别诊断的必要辅助方法。     

非何杰金淋巴瘤流式细胞术DNA含量分析的临床应用

非何杰金淋巴瘤(NHL)是一组高度异质性疾病,病理组织学分类和临床分期是评价预后的重要依据,但还远不能解释临床上遇到的复杂现象.近年来的研究证实NHL病变组织DNA含量的变化与预后明显相关.本文介绍NHL流式细胞术DNA含量分析的临床应用.综合文献报告细胞周期测定结果,123例对照组淋巴结(包括淋巴结反应性增生,淋巴结炎及原因不明的良性淋巴结肿大)     

Livin在非霍奇金淋巴瘤中表达的临床意义

本研究旨在探讨凋亡抑制因子Livin在非霍奇金淋巴瘤组织中的表达及其临床意义。应用免疫组织化学法检测Livin蛋白在30例非霍奇金淋巴瘤(non-Hodgkin′s lymphoma,NHL)患者及11例淋巴结反应性增生(reac-tive hyperplasia of lymph node)患者淋巴结中的表达,并应用real-time PCR对其中20例非霍奇金淋巴瘤、10例淋巴结反应性增生及4例正常人的淋巴结组织中Livin mRNA表达水平进行检测,分析livin蛋白和mRNA表达与NHL患者临床表现及其特征的相关性。结果表明,NHL患者淋巴结组织中Livin mRNA表达水平与正常淋巴结组织以及淋巴结反应性增生组织比较均显著增高(RQ中位数分别为12.4vs0.34和12.4vs0.61)(p0.01)。30例非霍奇金淋巴瘤中有16例Livin蛋白表达阳性,阳性率为53.3%;11例淋巴结反应性增生患者仅1例表达阳性,阳性率为9.1%;而且观察到Livin蛋白主要在细胞浆中表达,在胞核内极少见有表达。研究还显示,无论是LivinmRNA表达还是蛋白表达均与NHL的临床分期(p=0.023;p=0.009)、B症状(p=0.015;p=0.026)、血β2微球蛋白(β2-MG,p=0.031;p=0.012)及血清乳酸脱氢酶(LDH,p=0.037;p=0.007)密切相关,而两者的表达与年龄、性别及分型无明显相关性(p0.05)。结论:Livin mRNA及蛋白在NHL患者中表达增高,并与临床多项指标存在相关性,因此Livin表达水平对评价患者临床分期及预后可能具有重要的指导意义;对Livin在NHL中的作用机制的深入探讨将有助于揭示NHL的发生、发展及治疗的机理,并有望成为NHL未来治疗的重要突破点。     

Skp2和p27kip1蛋白在B细胞性非霍奇金淋巴瘤中的表达、定位及意义

目的 探讨B细胞性非霍奇金淋巴瘤(B-NHL)组织中Skp2和p27kip1蛋白的表达、定位及意义.方法 用免疫组织化学方法检测72例B-NHL和14例反应性增生淋巴结组织Skp2、p27kip1蛋白及细胞增殖指标Ki-67的表达情况,结合病理资料进行统计分析;用免疫荧光双标记技术检测淋巴瘤Raji细胞株Skp2和p27kip1蛋白的定位情况.结果 Skp2蛋白在B-NHL组织中的表达高于反应性增生的淋巴结(不含生发中心),在侵袭性B-NHL组织中的表达高于惰性组;p27kip1蛋白在B-NHL组织中的表达低于反应性增生淋巴结(不含生发中心),在侵袭性B-NHL组织中的表达低于惰性组;skp2蛋白主要在Raji细胞的胞浆中表达,p27kip1蛋白主要表达于胞核.结论 在B-NHL组织中,Skp2的高表达和p27kip1的低表达可能与其发生发展有关;skp2和p27kip1在Raji细胞中的亚细胞定位与其生物学功能一致.     

淋巴瘤白血病的病理类型与临床特征分析

目的 探讨淋巴瘤白血病(LML)患者的病理类型和临床表现.方法 按照2008年世界卫生组织(WHO)造血与淋巴组织肿瘤分类方案,回顾性分析692例非霍奇金淋巴瘤(NHL)患者中127例LML患者病理类型与临床特征.应用组织形态学、免疫组织化学、流式细胞术和骨髓检查结合临床资料进行研究.结果 15种NHL病理类型发生LML,其中发病率较高的NHL病理类型依次为B原始淋巴细胞淋巴瘤(B-LBL)、小淋巴细胞淋巴瘤(SLL)和T原始淋巴细胞淋巴瘤(T-LBL).LML的T和B细胞亚型分别为45例和74例.两组总体生存(OS)率差异有统计学意义(P＜0.01).81例患者初诊时已为LML,46例患者在病程1～88个月发展为LML,原发淋巴结内和淋巴结外者分别为96例和31例,LBL和急性淋巴细胞白血病、SLL和慢性淋巴细胞白血病有不同的发病形式和发展阶段.结论 LML并不少见.发生LML的病理类型有15种之多.其临床表现比较特殊,尤其是原发淋巴结外的LML更为独特.     

侵犯骨髓的非霍奇金淋巴瘤41例病理诊断探讨

目的 研究组织病理学在非霍奇金淋巴瘤(NHL)侵犯骨髓中的诊断价值.方法 骨髓活检标本观察组织形态,流式细胞学(FC)、免疫组化(IHC)进行免疫表型分析.结果 41例确诊为NHL病例中,B细胞淋巴瘤27例,T细胞淋巴瘤14例.侵犯骨髓的NHL分型:弥漫型15例(36.59%),混合型11例(26.83%),间质型9例(21.95%),结节型5例(12.20%),窦内型1例(2.44%).FC可对37例NHL(90.24%)分型,其余4例需结合病史及骨髓IHC结果 确定.15例同时行IHC检测,其中12例明确分型,其余3例(2例SLL/CLL,1例HCL)需结合FC进一步分型.1例DLBCL经FC仍不能分型需结合IHC确定.结论 骨髓病理组织形态、免疫表型在NHL侵犯骨髓的诊断方面具有重要价值,流式细胞学与免疫组化在免疫分型方面互为补充.     

sB7-H4在恶性淋巴瘤患者血清和淋巴瘤组织中的表达及其对淋巴瘤诊断和复查的价值

目的:分析可溶性B7-H4(sB7-H4)在恶性淋巴瘤患者血清和淋巴瘤组织中的表达及其对淋巴瘤诊断和复查的价值。方法:收集咸阳市中心医院83例恶性淋巴瘤患者的血清标本,其中初发恶性淋巴瘤患者69例为A组,包括霍奇金淋巴瘤患者11例为HL组,非霍奇金淋巴瘤患者58例为NHL组; 14例复发的患者为B组,另同期收集于本院体检合格者50例的血清标本,并作为对照组。用双抗体夹心酶联免疫吸附试验(ELISA)检测各组血清标本中的sB7-H4水平,并采用免疫组织化学法检测sB7-H4在恶性淋巴瘤组织和淋巴细胞反应性增生组织中的表达。结果:A组血清中sB7-H4的水平较B组和对照组均显著升高,而B组的水平又显著高于对照组(P 0. 05);在NHL组血清中sB7-H4的水平较HL组、对照组均显著升高,而HL组的水平又显著高于对照组(P 0. 05)。sB7-H4在淋巴细胞反应性增生组织中的表达均为阴性,而在恶性淋巴瘤组织的阳性表达率为47. 50%。结果提示,sB7-H4在恶性淋巴瘤组织中的阳性表达率较其在淋巴细胞反应性增生组织中的阳性率显著升高(P 0. 05)。结论:sB7-H4在恶性淋巴瘤患者血清和淋巴瘤组织中呈现高表达,其对淋巴瘤诊断和复查具有一定的价值。     

Cyclin A与PCNA在B细胞性非霍奇金淋巴瘤中的表达

目的 探讨细胞周期蛋白A(cyclin A)和增殖细胞核抗原(PCNA)在B细胞性非霍奇金淋巴瘤(B-NHL)中的表达及意义。方法 采用免疫组织化学法检测65例B-NHL及20例反应性增生淋巴结中cyclin A和PCNA蛋白的表达情况。结果 cyclin A和PCNA在B-NHL中的平均标记指数(1abel index,LI)明显高于反应性增生性淋巴结(生发中心以外的淋巴组织);侵袭性B-NHL中cyclin A的LI和PCNA的LI均高于惰性组;cyclin A与PCNA的表达成正相关。结论 cyclin A和PCNA的高表达与B-NHL的发生发展有关,cyclin A的LI与PCNA LI可以作为衡量B-NHL恶性程度的重要指标。     

Flow cytometric immunophenotyping including Bcl-2 detection on fine needle aspirates in the diagnosis of reactive lymphadenopathy and non-Hodgkin's lymphoma

BACKGROUND: Fine-needle aspiration (FNA) with immunophenotyping by immunocytochemistry (IC) on cytospins has recently received increased consideration in the diagnosis of lymphoma. The aim of our study was to establish the diagnostic value of a four-color flow cytometric (FCM) panel, including cytoplasmic Bcl-2, in cytologic diagnosis of malignant non-Hodgkin's lymphoma (NHL) and reactive lymphoid hyperplasia (RH). METHODS: We investigated 424 FNAs from 396 patients. FCM panel included lambda/kappa/CD19/CD5, CD23/CD10/CD20/CD19, CD4/CD7/CD8/CD3 and Bcl-2/CD10/CD19/CD3 in fluorescein isothiocyanate, phycoerythrin, and peridinin chlorophyll protein or a tandem conjugate of R-phycoerythrin and indodicarbocyanine and allophycocyanin. Bcl-2 expression was evaluated separately for gated B and T cells. RESULTS: In 97% of 172 RH samples, FCM was concordant with the diagnosis. FCM gave correct immunologic diagnosis in 95% of low-grade B-cell NHLs, 78% of high-grade B-cell NHLs, and 53% of T-cell lymphomas. Malignant B cells had higher Bcl-2 expression than did reactive B and T cells. This helped to establish a correct diagnosis especially in cases where no clear-cut monoclonality could be shown by kappa/lambda staining or where there was no expression of surface light chain. The highest Bcl-2 expression was found in follicular lymphomas. CONCLUSION: Our FCM panel allowed precise classification of NHL in FNA material in 89.5% of all samples. Bcl-2 staining can be recommended for primary differentiation between reactive hyperplasia and NHL.     

125例非霍奇金淋巴瘤患者的临床和预后分析

本研究旨在分析非霍奇金淋巴瘤（NHL）的临床资料，探讨影响预后的因素。对125例NHL病例进行了回溯性分析。结果发现，125例病例中B—NHL85例（68％），T—NHL35例（28％），不能分类的5例（4％）。B—NHL易于侵犯骨髓，而T—NHL多伴有B症状，乳酸脱氢酶（LDH）升高，临床分期晚，国际预后指数（IPI）评分高，且T—NHL的3年存活率明显低于B—NHL。生存分析显示，年龄、B症状、LDH水平、临床分期为预后相关因素，免疫分型对预后无显著性影响。骨髓侵犯的发生率为31．2％（39／125），在B—NHL多见。骨髓侵犯方式与年龄、B症状、LDH水平、T／B免疫分型无关，但弥漫型骨髓侵犯多伴有肝脾肿大，且总体生存率明显低于局灶型。结论：年龄、B症状、LDH水平和临床分期是影响NHL预后的主要因素，免疫分型尚不足以成为判断预后的独立因素，骨髓侵犯方式对预后有一定的提示意义。     

Diagnosis of lymphoproliferative disorders: experience of a single institution in the long-term follow-up of discordant cases

OBJECTIVE: To evaluate the usefulness of morphologic diagnosis, immunophenotyping and immunoglobulin (Ig) and T-cell receptor (TcR) gene rearrangement studies in the diagnosis of lymphoproliferative disorder. DESIGN: A retrospective cohort study with clinical follow-up of controversial cases. SETTING: Single institution, tertiary care centre. PATIENTS: All 273 patients whose lymphoid tissue samples were sent for molecular analysis by Southern blotting over a 4-year period. INTERVENTIONS: Patient reports were retrieved from the laboratory data system. Repeat assessment and clinical follow-up was done for discordant cases. OUTCOME MEASURES: Correlation between morphologic features and the results of immunophenotype and gene rearrangement studies of the samples. Value of the different tests in discordant cases. RESULTS: The 273 samples included 130 of non-Hodgkin's lymphoma (NHL), 23 of Hodgkin's disease, 80 of benign lymphoid hyperplasia, 16 of atypical lymphoid hyperplasia and other diagnoses. Of the 130 NHL cases diagnosed by morphologic study, 22 (17%) did not show gene rearrangement. Of the 80 morphologically benign samples, 4 (5%) showed gene rearrangement, and malignant disease developed later in those patients. Five (31%) of the 16 cases of atypical lymphoid hyperplasia showed gene rearrangement. Six of the remaining 11 cases had no detectable gene rearrangement, but hematologic malignant disease developed. No gene rearrangement was detected in Hodgkin's disease samples. One carcinoma showed gene rearrangement. Of the NHL group, 86% of the B cells and 50% of the T cells showed gene rearrangement. Seven samples showed both Ig and TcR gene rearrangement. CONCLUSIONS: Gene rearrangement analyses correlate highly with conventional morphologic diagnosis and phenotyping. The detection of gene rearrangement in lymphoid tissue has a high specificity (99%) and a reasonable sensitivity (83%) to the development of a lymphoproliferative disorder.     

慢性粒-单核细胞白血病合并结外非霍奇金淋巴瘤的临床特征分析

目的：探讨慢性粒-单核细胞白血病(chronic myelomonocytic leukemia, CMML)合并结外非霍奇金淋巴瘤（non-Hodgkin's lymphoma,NHL）患者的临床特征及其鉴别诊断。方法：回顾性分析2例CMML合并结外NHL患者的临床资料、影像学检查、病理形态学及免疫组织化学（免疫组化）标志特征,并复习国内外相关文献进行探讨。结果：2例CMML合并结外NHL患者均为老年女性,因乏力或出现相应部位肿瘤侵犯症状就诊。例1患者诊断为CMML后发生NHL,例2患者则同时诊断为CMML和NHL。实验室检查示,2例患者的外周血白细胞计数增高,以单核细胞为主;乳酸脱氢酶升高;骨髓增生活跃伴有病态造血;细胞免疫表型为CD14⁺ CD64⁺;染色体荧光原位杂交分析BCR-ABL为阴性(-)。PET/CT 检查显示 2例患者均未发现骨髓高代谢灶,例1患者表现为小肠异常高代谢,最大标准化摄取值（maximum standard uptake values,SUVmax）值为12.60;例2患者表现为肺内异常高代谢,SUVmax值为6.72。病理检查提示2例患者均为B细胞来源的NHL;免疫组化检查结果提示,LCA、CD20、CD79a、Bcl-6、Ki-67均为阳性,CD5、CD10、CD3、CylinD1、CD56、CD23、CD21阴性。结论：CMML合并结外NHL罕见,患者缺乏特异性的临床表现,而结合实验室、影像学检查及组织病理、免疫组化检查进行分析,可提高其诊断的准确率。     

Immunophenotyping by flow cytometry of fine needle aspirates in the diagnosis of lymphoproliferative disorders: A retrospective study.

In order to determine the value of flow cytometric (FCM) immunophenotyping of fine-needle aspirates (FNA) in the diagnosis and classification of lymphoproliferative diseases, 61 tissue samples were studied and compared with the cytologic/histological results. In vivo and ex vivo FNA biopsy yielded the material for FCM, which comprised an extensive number of lymphoid cell markers. In all but three cases sufficient cells were collected. Overall, malignancy was diagnosed in 33 cases from a total of 47 (70.2%), and in the remaining cases malignancy was not detected. Eleven cases were correctly diagnosed as reactive processes (11/11). There were no false positive cases of malignancy, as diagnosed by FCM-FNA. The best accuracy was achieved in the low-grade B-cell lymphomas and lymphoblastic lymphoma/leukemia. We conclude that in a significant number of cases, FCM-FNA permits the separation between lymphoid malignancies and reactive processes without false positive results. It was found to be particularly useful in the differential diagnosis of mantle-cell and small-lymphocytic lymphoma and in the identification of lymphoblastic lymphoma/leukemia.     

伴有明显淋巴滤泡的非特指外周T细胞淋巴瘤

目的　探讨伴有明显淋巴滤泡增生的非特指外周T细胞淋巴瘤的形态学特点和免疫表型。方法　对 3例特殊形态的外周T细胞淋巴瘤进行形态学观察 ,免疫组化EnVision法检测CD3、CD4 5RO、CD4 3、CD2 0、CD79a、cyclinD1、bcl 2、CD4、CD8、S 10 0等抗体 ,并用PCR方法进行T细胞受体(TCR)基因重排检测。结果　 3例患者均以全身浅表淋巴结肿大为主要表现 ,初发部位为颈部和颌下 ,发病过程中出现低热及全身皮疹 ,并有肝、脾肿大。形态学的显著特点为淋巴滤泡增生 ,套区消失 ,边缘区有中等大小、胞浆透亮的瘤细胞浸润生长。免疫表型标志为T细胞淋巴瘤。基因检测有TCRγ基因阳性条带。结论　部分非特指外周T细胞淋巴瘤可伴有明显淋巴滤泡增生 ,应注意与淋巴结反应性增生、滤泡性淋巴瘤、边缘区淋巴瘤及套区淋巴瘤等鉴别     

Detection of T-regulatory cells has a potential role in the diagnosis of classical Hodgkin lymphoma

Previous studies have demonstrated an increase in T-regulatory cells in the involved lymph nodes and peripheral blood of patients with Hodgkin lymphoma. Our study examined whether the detection of T-regulatory cells by flow cytometry could distinguish classical Hodgkin lymphoma (CHL) from benign cases and B-cell non-Hodgkin lymphomas (B-NHL). We measured CD4, CD25, and CD152 in 14 CHLs, 2 nodular lymphocyte-predominant Hodgkin lymphomas, 31 B-NHLs, and 54 benign cases. All T-regulatory cell parameters, including percent lymphocytes CD4+/CD152+ and CD4+/CD25+/CD152+, and mean and median CD152 expression in CD4+/CD25+ lymphocytes, were higher in CHL than in B-NHL and benign. Mean CD152 in CD4+/CD25+ lymphocytes distinguished CHL from benign with 79% sensitivity and 100% specificity, and from B-NHL with 71% sensitivity and 90% specificity. Overall, our results show that T-regulatory cells are increased in CHL and their detection may be a useful tool in differentiating CHL from other entities.     

Flow cytometric analysis of cytologic specimens in hematologic disease

Cytologic evaluation of body fluids and fine needle aspirations (FNA) is frequently required in patients with hematologic diseases. In this study we have correlated immunophenotyping and DNA analysis by flow cytometry with cytologic findings and tissue biopsies from 20 patients with body fluid specimens and 5 with FNA. Nineteen of 25 cases, including all FNA cases, had an immunophenotype consistent with malignancy: 12 monoclonal B-cell lymphomas, 2 T-cell lymphomas, 2 T-ALL and 3 non-T-ALL. By cytologic examination, 14 of these 19 cases were positive for malignant cells, 2 suspicious and 3 negative; the latter 5 cases, including 2 FNA cases, had small monoclonal B-cell populations detected by flow cytometry. Six cases had a benign immunophenotype; cytologic examination was benign in 4 of these and suspicious for lymphoma in 2. Our results show the feasibility of using flow cytometry to evaluate body fluids or FNA and demonstrate that small malignant populations that may be missed by routine cytology can be detected by flow cytometry.     

Multiple monoclonal B cell expansions and c-myc oncogene rearrangements in acquired immune deficiency syndrome-related lymphoproliferative disorders. Implications for lymphomagenesis

AIDS (acquired immune deficiency syndrome) and ARC (AIDS-related complex) are associated with a spectrum of lymphoproliferative disorders ranging from lymphadenopathy syndrome (LAS), an apparently benign polyclonal lymphoid hyperplasia, to B cell non-Hodgkin's lymphoma (B-NHL), i.e., malignant, presumably monoclonal B cell proliferations. To gain insight into the process of lymphomagenesis in AIDS and to investigate a possible pathogenetic relationship between LAS and NHL, we investigated the clonality of the B or T lymphoid populations by Ig or T beta gene rearrangement analysis, the presence of rearrangements involving the c-myc oncogene locus, and the presence of human immunodeficiency virus (HIV) sequences in both LAS and B-NHL biopsies. Our data indicate that multiple clonal B cell expansions are present in a significant percentage of LAS (approximately 20%) and B-NHL (60%) biopsies. c-myc rearrangements/translocations are detectable in 9 of our 10 NHLs, but not in any of the LAS cases. However, only one of the B cell clones, identified by Ig gene rearrangements carries a c-myc gene rearrangement, suggesting that only one clone carries the genetic abnormality associated with malignant B cell lymphoma. Furthermore, the frequency of detection of c-myc rearrangements in AIDS-associated NHLs of both Burkitt and non-Burkitt type suggest that the biological alterations present in AIDS favor the development of lymphomas carrying activated c-myc oncogenes. Finally, our data show that HIV DNA sequences are not detectable in LAS nor in NHL B cell clones, suggesting that HIV does not play a direct role in NHL development. Taken together, these observations suggest a model of multistep lymphomagenesis in AIDS in which LAS would represent a predisposing condition to NHL. Immunosuppression and EBV infection present in LAS can favor the expansion of B cell clones, which in turn may increase the probability of occurrence of c-myc rearrangements leading to malignant transformation.