Diminished expression of CD19 in B-cell lymphomas

BACKGROUND: CD19 is expressed on most B-cell lymphomas; however, the frequency and types of B-cell lymphomas with low-level expression of CD19 are not well characterized. METHODS: We reviewed flow cytometric histograms specifically for decreased CD19 expression on 349 cases analyzed by the Flow Cytometry Laboratory at University Hospitals of Cleveland (Cleveland, Ohio). Results of flow cytometry were correlated with the morphologic diagnosis. RESULTS: Of the cases reviewed, 125 (36%) showed a visible decrease in CD19 expression compared with normal B lymphocytes. Decreased CD19 expression was noted in 79% of follicular lymphomas (27 of 34), 36% of small lymphocytic lymphomas/chronic lymphocytic leukemias (82 of 228), 31% of mantle cell lymphomas (4 of 13), 24% of diffuse large B-cell lymphomas (8 of 33), and 13% of marginal zone B-cell lymphomas/lymphoplasmacytoid lymphomas (4 of 30) and was not observed in any Burkitt lymphoma (0 of 5) or hairy cell leukemia (0 of 6). Decreased CD19 expression was significantly more frequent in follicular lymphomas than in other lymphoma subtypes (P < 0.001). No significant difference was observed in the frequency of decreased CD19 expression based on histologic grade of follicular lymphoma. CONCLUSIONS: Diminished expression of CD19 expression occurs frequently in B-cell lymphomas, in particular follicular lymphoma, and may be helpful in identifying B-cell lymphoma cells in complex cell mixtures such as bone marrow specimens.     

Flow fluorimetry in differential diagnosis of diffuse large B-cell lymphoma

AIM: To analyse immunophenotype of diffuse large B-cell lymphoma (DLBCL) with flow cytometry. MATERIAL AND METHODS: Combinations of antibodies against the following antigens were used: CD3/ CD19/CD45, CD5/CD19/CD38, CD19/CD10/CD23, CD4/CD8/CD3, kappa/lambda/CD19, CD25/CD20/FMC7; CD43/CD22/CD20; CD79a/Ki-67/CD3; cytoplasmic kappa/lambda. The analysis was made on flow fluorimeter FacsCalibur using computer program CellQuest (Beckton Dickenson, USA). RESULTS: Specific coexpression of markers is not detectable in DLBCL, in the greatest degree the phenotype corresponds to lymphoma from the cells of the marginal zone. The study of cells with maximal direct light diffusion provides more precise assessment of clonality and proliferative potential of tumor cells than the analysis of the whole lymphocytic polygon. The proliferative index in 33 cases of DLBCL varied in the range 10-60%. CONCLUSION: Flow cytometry in most DLBCL cases allows identification of B-cell clonality, more precise assessment of a proliferative potential of the tumor.     

Guess what: Chronic 13q14.3+/CD5-/CD23+ lymphocytic leukemia in blood and t(11;14)(q13;q32)+/CD5+/CD23- mantle cell lymphoma in lymph nodes!

We report a case of a patient with two B-cell lymphoproliferative disorders: CD5(-)/CD23(+) B-cell chronic lymphocytic leukemia and CD5(+)/CD23(-) mantle cell lymphoma. These disorders were diagnosed simultaneously based on flow cytometry, immunohistochemistry, fluorescence in situ hybridization, and polymerase chain reaction-based molecular studies. The B-cell lymphocytic leukemia clone predominated in the blood and bone marrow, whereas the mantle cell clone predominated in lymph nodes.     

Utility of flow cytometry in subtyping composite and sequential lymphoma.

Composite lymphoma (CL) is defined as more than one distinct lymphoma variant occurring in the same anatomic site, and sequential lymphoma (SL) is defined as different lymphoma variants occurring at different sites or at different times in the same patient. The utility of flow cytometry immunophenotyping in evaluating CL and SL has only been investigated in a few single-case studies. To further define the utility of flow cytometry in evaluating these tumors, records were searched at two institutions. Cases representing high-grade progression of low-grade lymphoma were excluded. For each CL/SL, clinical data was obtained and morphology was evaluated in routinely processed H&E-stained tissue sections. Tumor components were subtyped using revised European-American classification (REAL) criteria. Follicle center components were graded using modified Rappaport criteria. Immunophenotype was determined using two-color flow cytometry and paraffin-section immunostains. Four cases were identified. Case 1, nodal follicle center, follicular, grade III plus marginal zone CL, showed two discrete populations of monoclonal B-cells that differed in their expression of CD10. Case 2, cutaneous lymphoplasmacytoid lymphoma followed by mesenteric non-Hodgkin's lymphoma (lymphoplasmacytoid plus follicle center, follicular, grade III) plus Hodgkin's disease CL, showed CD5-/CD10-/CD19+/kappa+ cells by flow cytometry in both tissue samples. The Hodgkin's disease component showed CD3-/CD15-/CD20-/CD30+ Reed-Sternberg cell variants in paraffin-section immunostains. Case 3 represented nodal follicle center lymphoma, follicular, grade I (CD3-/CD5-/CD10-/CD19+/kappa+) followed by cutaneous anaplastic large T-cell lymphoma (CD2+/CD4+/CD5+/CD19- cells with partial expression of CD3 and CD7). Case 4 represented cutaneous follicle center lymphoma, follicular, grade I (CD5-/CD10+/CD19+/CD23+/lambda+) followed by bone marrow B-cell small lymphocytic lymphoma (CD5+/CD10-/CD19+/CD23+/kappa+). Results show that flow cytometry is a potentially useful adjunct in characterizing CL and SL.     

淋巴结细针穿刺标本的流式细胞术分析

目的 评价淋巴结细针穿刺标本流式细胞术分析方法在诊断淋巴结病变,以及在恶性淋巴瘤和淋巴结反应性增生鉴别诊断中的应用价值.方法 对99份疑为淋巴结病变的淋巴结针吸标本涂片进行常规细胞学分析,并结合病理活组织检查确诊分型.使用三色标记的流式细胞术方法分析针吸标本中各细胞免疫表型(CD3、CD3、CD4、CD5、CD10、CD19、CD20、CD23、CD45、K、λ、FMC7、 CD34),确定标本中各细胞组成及有无异常表型细胞.对于淋巴瘤病例,则按照WHO分型标准,根据免疫表型进一步确定其亚型,并对各类病例流式细胞术分型结果和细胞学结果进行比较.结果 99份标本中,细胞涂片检出淋巴瘤40例,转移癌29例,反应性增生、坏死、结核30例,有2例非霍奇金淋巴瘤(NHL)被误诊为反应性增生;对其中18例NHL进行了病理活组织检查,其中包括B淋巴细胞非霍奇金淋巴瘤(B-NHL)16例,T淋巴细胞非霍奇金淋巴瘤2例.流式细胞术分析结果显示,99份标本中检出淋巴瘤35例(淋巴母细胞淋巴瘤4例,T淋巴细胞病变1例,其余30例为B-NHL);有28份B-NHL检测到K或λ轻链限制性表达,其K:λ或λ:K大于3:1,B淋巴细胞所占比例为(73.2±27.2)%,其中26份能根据免疫标志物的表达确定其亚型.经病理活组织检查确定为B-NHL的16份标本中,仅2例滤泡淋巴瘤与流式细胞术分型结果不一致.对于反应性增生及转移癌等标本,流式细胞术分析未查见异常淋巴细胞,其k:λ或λ:k均小于3:1.结论 淋巴结细针穿刺标本的流式细胞术分析有助于淋巴结病变的诊断和鉴别诊断,并可确定NHL的亚型.     

Circulating levels of soluble CD23 reflect clinical and biological features of leukemic B-cell chronic lymphoproliferative disorders

Summary One hundred and twenty-four sera, from patients with various leukemic B-cell chronic lymphoproliferative diseases were investigated at diagnosis by ELISA for their soluble CD23 content. Immunophenotyping was carried out in all patients, and in a selected subset the mean number of membrane-bound CD23 molecules per cell was also investigated. Seventy-three patients had typical B chronic lymphocytic leukenia, 41 leukemic B-cell disorders with atypical morphological and/or immunophenotypic features, 5 had low-grade follicular cell lymphoma in the leukemic phase, and 5 had splenic lymphoma with villous lymphocytes Soluble CD23 levels were significantly higher than in normal sera (mean±SD: typical B chronic lymphocytic leukemia 3,650±4,654 U/ml, atypical B chronic lymphocytic leukemia 3,440±4,671 U/ml, follicular cell lymphoma 3,200±1,511 U/ml, splenic lymphoma with villous lymphocytes 8,236±7,294 U/ml, controls 137±128 U/ml;P<0.001). More advanced Rai's stages were related to higher soluble CD23 levels (P<0.01), both in typical and atypical B chronic lymphocytic leukemias, the highest levels and the best correlation with the absolute number of circulating CD19+ cells (r=0.50) being observed in the typical form. The number of membrane-bound CD23 molecules per cell was significantly higher in typical than in atypical B chronic lymphocytic leukemias (mean number 156,727±94,668 vs. 12,010±10,643,P<0.001). Our data suggest that soluble CD23 levels correlate with the clinical and biological features of leukemic B-cell lymphoproliferative disorders.     

Spontaneous remission of B-cell chronic lymphocytic leukemia associated with T lymphocytic hyperplasia in bone marrow

Spontaneous remissions in B-cell chronic lymphocytic leukemia (B-CLL) are rare and none of them has been studied with immunophenotyping (by flow cytometry and immunohistochemistry) and genotyping. In this patient, studied after spontaneous remission had occurred, there was a residual T-lymphocytic hyperplasia in the bone marrow with a normal CD4:CD8 ratio. Absolute CD4 and CD8 counts and CD4:CD8 ratio in the peripheral blood were normal. Flow cytometry revealed no B-CLL cells in the peripheral blood and less than 2% B-CLL cells in the bone marrow.     

脾脏套细胞淋巴瘤临床病理分析

目的 :探讨脾脏套细胞淋巴瘤 (SMCL)的临床病理特征、诊断和鉴别诊断。方法 :对 3例SMCL进行临床、组织病理和免疫组化分析。结果 :男性 1例 ,女性 2例 ,平均年龄 5 9 67岁。 3例除脾脏肿大外 ,还伴有骨髓及外周血的侵犯。镜下特征为形态单一的瘤细胞呈弥漫性和 /或结节性生长 ,瘤细胞免疫表型特征为CD5 ( +) ,cyclinD1 ( +) ,IgD( +)。 结论 :诊断SM CL必需结合组织学特点和免疫表型 ,还需与滤泡性淋巴瘤、小淋巴细胞性淋巴瘤、脾边缘区B细胞淋巴瘤鉴别。     

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.     

肝血管内大B细胞性淋巴瘤2例临床病理观察

目的探讨肝血管内大B细胞性淋巴瘤的临床病理特征、诊断与鉴别诊断、治疗及预后。方法回顾性分析2例肝血管内大B细胞性淋巴瘤患者的临床资料、组织病理学形态和免疫组化结果。结果光镜下肝窦内和小血管内可见较多具有明显异型性的淋巴样细胞浸润,汇管区可见慢性炎细胞浸润,亦可见少许异型淋巴细胞样细胞,未见明确纤维化。免疫组化示CD20、PAX5弥漫(+),CD3散在少许(+),Ki-67阳性率为70%,AE1/AE3、CD117和CD56均(-);其中例1 CD5弥漫(+)。结论血管内大B细胞性淋巴瘤是一种具有高度侵袭性的结外弥漫性大B细胞性淋巴瘤的亚型,由于该病临床表现多样及不典型性,造成了部分患者的诊断困难,因此,掌握临床病理及免疫组化特征对该病的诊断和鉴别诊断具有重要意义。     

恶性血液病细胞中tankyrase表达与端粒酶活性关系研究

为研究以白血病为主的恶性血液病细胞中端粒酶活性正向调控基因tankyrase的表达与端粒酶活性的关系并初步探讨tankyrase对端粒酶活性调控的机理和意义 ,以实时定量RT PCR技术对髓细胞系白血病细胞株K5 6 2 ,HL 6 0 ,U937,NB4 ,THP 1,HEL ,Dami,T淋巴细胞性白血病细胞株 6T CEM ,Jurkat和B细胞淋巴瘤细胞株Raji中tankyrase的表达进行检测 ,同时检测端粒酶逆转录酶hTERT的表达确定端粒酶活性 ,并以经磁珠分离的正常人CD3 ,CD19 和CD33 细胞和 10份正常人骨髓单个核细胞做对照。结果发现 :tankyrase在恶性血液病细胞株中的表达明显高于正常对照 (U =19,P <0 .0 1) ,其中髓系白血病细胞株中的表达高于正常人CD33 细胞 ,T淋巴细胞性白血病细胞株和B细胞淋巴瘤细胞株中的表达分别高于正常人CD3 和CD19 细胞。髓系恶性血液病细胞株tankyrase的表达 (0 .0 0 32± 0 .0 0 10 )明显低于淋系恶性血液病细胞株的表达 (0 .0 12± 0 .0 0 16 ) (F =2 3,P <0 .0 1)。Tankyrase与hTERT的表达呈正相关 (相关系数为 0 .395 ,P <0 .0 5 )。结论 :tankyrase在恶性血液病细胞株中呈高表达 ,与端粒酶活性呈正相关 ,提示tankyrase可能是恶性血液病中端粒酶活性增高的原因之一。     

慢性淋巴细胞系统白血病免疫表型分析

目的：研究国内慢性淋巴细胞系统折血病的免疫表型特点。方法：采用单参数和多参数流式细胞术分析了163例慢性淋巴细胞系统白血病的免疫表型。结果：71.8％（117/163）患者共表达CD5和B细胞标志。采用WHO引用的计分系统，将病例分为B-慢性淋巴细胞白血病（B-CLL），毛细胞白血病（HCL）和其他B淋巴细胞增殖性疾病。典型的B-CLL表达CD5、CD23、CD20、CD19、HLA-DR，但仍有部分患者表达CD22、CD11c、CD25和FMC7。CD103似为HCL最特异的标志。但仅仅依靠免疫表型难以鉴别非典型B-CLL、B细胞-幼淋巴细胞白血病（B-PLL）和外套细胞淋巴瘤（MCL），细胞遗传学或分子生物学检查将有助于鉴别诊断。以同一标本中残存的正常淋巴细胞为内参照，计算前向角光散射（FSC）指数和抗原表达指数，可定量地表示细胞的大小和抗原表达的强度，使不同的标本具有可比性。结论：免疫表型分析是诊断慢性淋巴细胞系统白血病非常有用的依据。     

子宫体原发性弥漫性大B细胞淋巴瘤临床病理观察

目的探讨子宫体原发性弥漫性B细胞淋巴瘤的病理诊断与鉴别诊断要点。方法对2例子宫体原发性弥漫性B细胞淋巴瘤进行临床病理特点观察,使用LCA、CD30、CD79a、CD68、CD45RO、CD3、CD20和CEA等免疫组化染色辅助诊断,并文献复习。结果子宫体原发性弥漫性B细胞淋巴瘤临床诊断困难,镜下病理形态多样,可见子宫内膜和/或肌层内淋巴样细胞增生,免疫组化B细胞标记（＋）。结论子宫原发性弥漫性B细胞淋巴瘤少见,临床易漏诊且侵袭性较高,因此准确、完整的病理诊断对预后治疗十分重要,免疫组化有辅助诊断作用。     

鼻咽部良性淋巴组织增生209例临床病理分析

目的总结鼻咽部良性淋巴组织增生（NBLA）的临床病理特点、诊断及鉴别诊断要点,提高对NBLA的认识和病理诊断水平。方法观察209例NBLA的临床及组织病理学,用免疫组化EnVision法检测60例病变组织中浸润的淋巴细胞、雌激素（ER）和孕激素（PR）的表达情况,用原位杂交方法对16例病变组织进行了EBER原位检测。结果镜下可将NBLA分为滤泡型（188／209）及弥漫型（21／209）;黏膜固有层浅层可见浆细胞灶状或带状浸润占56％（117／209）,淋巴上皮样病变占99％（207／209）,血管壁浸润占34．9％（73／209）。免疫组化：滤泡型CD79a和CD20阳性细胞集中于套区及生发中心,CD10阳性细胞位于生发中心,bcl-2阳性细胞主要位于套区,CD3、CD45RO阳性细胞主要分布于弥散淋巴组织。弥漫型CD79a和CD20阳性细胞与CD3和CD45RO阳性细胞混杂分布,bcl-2阳性细胞弥漫分布,CD10（-）。淋巴上皮样病变内浸润的淋巴细胞主要是CIY20和CD79a（＋）,浸润血管壁的淋巴细胞以CD3和CD45RO（＋）为主。17例（17／60）个别细胞ER弱（＋）,PR均（-）。16例EBER均（-）。结论NBLA镜下可分为滤泡型和弥漫型,弥漫型应注意与多种B细胞淋巴瘤鉴别,其发病与病变部位的ER、PR及EB病毒感染无关。     

Molecular and flow cytometry characterization during the follow-up of three simultaneous lymphoproliferative disorders: hairy cell leukemia, monoclonal B-cell lymphocytosis, and CD4(++) /CD8(+/- dim) T-large granular lymphocytosis--a case report

The simultaneous diagnosis of hairy cell leukemia and monoclonal B-cell lymphocytosis with the characteristics of "indolent" chronic lymphocytic leukemia is rare but not unknown. However, an association with a third clonal lymphoproliferative disorder has not previously been described. We report the simultaneous presence of hairy cell leukemia, monoclonal B-cell lymphocytosis, and alpha beta CD4(++) /CD8(+) T-cell large granular lymphocytosis in a 63-year-old man. After the diagnosis, the three lymphoproliferative disorders (i.e., two of B-cell lineage and one of T-cell lineage) were characterized by analysis of multiple sequential bone marrow and peripheral blood samples using flow cytometry and molecular techniques. We discuss these findings in the context of chronic antigen stimulation, immunosuppression, and apoptotic pathway alterations, which might be implicated in the accumulation of these abnormal clones in the same patient. Because the phenotype of the three clones is compatible with fully differentiated B lymphocytes (consistent with a postgerminal origin) and T-CD4(++) cells, we favor the possibility of an antigen-driven mechanism and a dysregulation of homeostatic apoptosis in this patient.     

用流式细胞术从B淋巴增殖性疾病中鉴别脾边缘带淋巴瘤

脾边缘带淋巴瘤（Splenicmarginalzonelymphoma,SMZL）是一种相对少见的原发于脾脏的惰性B淋巴增殖性疾病,表现为外周血淋巴细胞计数和／或比例增高、脾大,而外周浅表淋巴结往往不大,需要应用诊断性脾切除病理检查得以确诊,但患者多数不能接受,早期诊断困难。本研究探索以流式细胞术（flowcytometry,FCM）为主的诊断途径的可行性。选取6例疑诊SMZL患者为研究对象,同时以10名骨髓健康供者及确诊的10例慢性淋巴细胞白血病（CLL）、3例毛细胞白血病（HCL）、3例淋巴浆细胞淋巴瘤／华氏巨球蛋白血症（LPL／wM）初诊患者为对照。对所有患者及对照组进行骨髓FCM免疫分型,所选抗体组合包括CD45、CD5、CD10、CD19、CD20、CD22、CD23、CD25、CD103、CD11c、CD123、κ、λ、CyclinD1等,同时结合骨髓细胞形态学检查。结果表明：6例疑诊sMzL患者表现为淋巴细胞增高和脾大。因外周淋巴结无肿大6例患者均未行淋巴结活检,仅1例患者行病理诊断性脾切除。经骨髓FCM免疫表型分析,除健康供者外均可发现骨髓中存在不同程度的异常成熟B细胞克隆,并通过CD5、CD10表达与否结合其他表型进一步与其他B细胞肿瘤鉴别。结果6例SMZL患者均CDl9＋、CD20＋,而CD10-,4例患者CD5-,2例CD5＋,而CD23、CD38、ZAP-70、CD11c、CD103、CD123、CyclinD1均不表达。骨髓细胞形态学检查可见有短绒毛的异常淋巴细胞,结合临床特征将6例患者诊断为SMZL。1例患者因合并脾功能亢进需要行脾切除,术后病理证实该病。10例CLL主要表达CD5、CD23之外;在3例HCL除表达CD11c、CD103、CD123,形态学上更有典型的”毛”;3例LPL／WM具有轻链限制性表达、IgM显著升高、浆样淋巴细胞增多的特点。结论：FCM免疫表型分析结合骨髓形态学可作为临床诊断SMZL有力的工具。     

Clinical significance of monoclonal B cells in cerebrospinal fluid

BACKGROUND: Morphologically malignant lymphocytes in the cerebrospinal fluid (CSF) are highly suggestive of central nervous system involvement by lymphoid malignancy. Although flow cytometry is increasingly used to detect a monoclonal B-cell population in the CSF, the significance of this finding in the absence of morphologically identifiable malignant cells is unknown. METHODS: We reviewed CSF flow cytometric results in 32 patients studied at a single institution over 5 years and identified patients who had monoclonal B-cells in the CSF. Clinical presentation and course were reviewed. RESULTS: Twelve patients had a monoclonal B-cell population in the CSF, but only three had clinical evidence of malignant CNS disease. Of the other nine patients, 4 had nonmalignant neurologic disease and five had a lymphoproliferative disorder: chronic lymphocytic leukemia (n = 4) and mantle cell lymphoma (n = 1). In patients who had chronic lymphocytic leukemia and mantle cell lymphoma, the monoclonal B-cell population was small and had an immunophenotype identical to that of circulating malignant B cells. None of these nine patients developed clinical evidence of malignant CNS involvement during follow-up. CONCLUSION: In patients who have indolent B-cell malignancies, the presence of monoclonal B cells in the CSF may not be diagnostic of clinically significant CNS involvement by a lymphoid malignancy.     

Effect of interferon-alpha on CD20 antigen expression of B-cell chronic lymphocytic leukemia

Chimeric CD20 monoclonal antibody as alternative therapy in relapsed low-grade non-Hodgkin's lymphoma (NHL) has produced responses in nearly 50% of patients. Augmenting CD20 expression on tumor cells and/or inducing its expression may increase the cell kill and effectiveness of antibody therapy. Peripheral blood lymphocytes from 19 patients with B-cell chronic lymphocytic leukemia (B-CLL) were incubated in vitro in the presence of interferon-alpha (IFN-alpha) (500 U/ml and 1,000 U/ml) for 24 and 72 hours. The effect on CD20 expression was studied by flow cytometry. The differences in the percentage positivity, the mean fluorescence intensity (MFI), and the product of percentage positivity and MFI were used to assess upregulation. There was a significant upregulation of CD20 expression on B cells seen at both concentrations after 24-hour priming (p < 0.01). B-CLL cells cultured for 72 hours in the presence of IFN-alpha also showed upregulation of CD20 expression; however, the degree of upregulation was much lower than that seen at 24 hours. There was no statistically significant increase in CD20 antigen expression on normal lymphocytes following cytokine exposure. These results suggest that IFN-alpha priming may augment the effectiveness of antibody therapy by directly upregulating CD20 antigen expression in addition to its indirect action through effector cells of the host.     

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 features of angioimmunoblastic T-cell lymphoma

BACKGROUND: The immunophenotypic features of angioimmunoblastic T-cell lymphoma (AILT) have not been well described. METHODS: We retrospectively reviewed our institutional experience with the flow cytometric features of 16 cases of AILT. RESULTS: Multiparameter flow cytometry was able to identify a distinct population of immunophenotypically aberrant T cells in 15 of 16 cases. In 13 lymph node specimens, the neoplastic cells ranged from 1.9 to 87% (median 23%) of cells. The ratio of reactive to neoplastic T cells ranged from 0.01 to 20 (median 1.5); reactive T cells outnumbered neoplastic in 9/13 (69%) cases. The neoplastic populations expressed CD2, CD4, CD5, and CD45RO in all cases, lacked expression of CD8 and CD56 in all cases, and showed negative or dim surface CD3 in most cases. CD10 was expressed by the neoplastic populations in 11 of 14 cases at diagnosis; in 3 of these 11 only a subpopulation of the neoplastic cells was CD10(+). CD10 tended to be absent on neoplastic cells in staging bone marrows. The neoplastic population in all but one of the 15 positive cases possessed multiple immunophenotypic abnormalities and these were generally retained during the follow-up analyses of several cases. CONCLUSIONS: These results indicate the potential utility of flow cytometry in the diagnosis and follow-up of AILT.