Cluster differentiation antigen expression, proliferative activity and clinical stage in centroblastic centrocytic lymphomas

Using a panel of B-cell antibodies recognizing clusters of leucocyte differentiation antigens, immunostaining patterns of eight reactive lymph nodes and 28 centroblastic/centrocytic and centrocytic lymphomas have been studied. Centroblastic/centrocytic and centrocytic lymphomas retained many of the B-cell differentiation antigens and neoplastic follicles partially recapitulated the staining patterns observed in reactive follicles. Centrocytic lymphomas usually expressed a heavy chain mantle zone-like phenotype. Nearly one-half of follicular lymphomas showed extension of neoplastic cells into interfollicular areas as evidenced by positivity for CD10 (common acute lymphoblastic leukaemia) and/or CD9 (immature B-cell) and CD23 (B-blast cell) antigens. Cases showing interfollicular involvement also manifested considerable phenotypic heterogeneity. Light chain restriction could not be used to determine interfollicular involvement because of the presence of many non-neoplastic cells. Most follicular lymphomas retained a polyclonal mantle around at least some neoplastic follicles and in no case was a monoclonal mantle seen. Most lymphomas (16/21) were diploid when examined by flow cytometry. Diploid tumours exhibiting interfollicular lymphomatous involvement had high proliferation (S + G2) fractions and these lymph nodes were usually derived from patients with widespread disease. Tumours containing a high percentage of cells in the G0/G1 phases displayed fewer B-cell differentiation antigens than tumours with low G0/G1 fractions.     

CD30 expression in non-Hodgkin''s lymphoma

The CD30 antigen has been reported as the immunophenotypic hallmark of a recently described category of non-Hodgkin's lymphoma, termed anaplastic large cell lymphoma. From a series of approximately 500 lymphomas, 17 cases showing typical anaplastic features have been identified. They were strongly labelled by monoclonal antibodies recognizing CD30 (Ki-1 or BerH2). However, 36 other lymphomas, mainly high-grade, of non-anaplastic cytology also expressed CD30, either diffusely or focally, with a staining pattern identical to that seen in anaplastic large cell lymphomas. This clearly suggests that such lymphomas cannot be identified solely on the basis of being high-grade non-Hodgkin's lymphomas showing CD30 positivity. From the present results, the distinction between the anaplastic and non-anaplastic types would be better made with antibodies to epithelial membrane antigen than to CD30. Clinical data, available for 48 of the patients (16 with anaplastic large cell lymphomas and 32 with non-anaplastic) revealed no significant differences with regard to age at presentation, sex or clinical signs. A short-term follow-up study of 25 patients revealed that for the first 2 years after diagnosis there were no significant differences in patient survival between anaplastic large cell lymphoma, other CD30+ high-grade lymphomas and all high-grade non-Hodgkin's lymphomas considered together. These findings, which must be confirmed by larger studies, suggest that in a general lymphoma clinic there is probably little justification for differentiating anaplastic large cell lymphomas or CD30+ lymphomas from other high-grade non-Hodgkin's lymphomas.     

Activation and differentiation antigen expression in B-cell non-Hodgkin's lymphoma

In an attempt to establish whether extended immuno-phenotyping allows more accurate definition of subgroups of B-cell non-Hodgkin's lymphoma (NHL) we have stained a series of 145 cases with a large panel of monoclonal antibodies that recognize B-cell differentiation and activation antigens. No antigen was expressed by all cases. The B-cell histogenesis in many cases could be confirmed only by using a panel of immunoglobulin and pan B-cell markers. There was marked phenotypic heterogeneity within and between major groups of B-cell NHL as delineated by the Kiel classification although the differentiation antigens CD5 (lymphocytic and centrocytic NHL) and OKT10 (plasma cell tumours) were more often expressed by certain morphological groups. The activation antigens 4F2 and transferrin receptor were expressed more strongly and more often by high grade NHL but other activation antigens (CD23 and CD25) were not more frequently associated with these tumours. Extended phenotyping may be of value in improving the understanding of biological abnormalities and processes involved in B-cell NHL, but we conclude that a limited panel of markers (CD3, CD5, CD22, CD45, IgM, kappa, and lambda) should be sufficient for routine diagnosis and classification of most cases.     

Flow cytometric detection of B-clonal excess in fine needle aspirates for enhanced diagnostic accuracy in non-Hodgkin''s lymphoma in adults

Fine needle aspiration (FNA) cytology is a valuable aid to diagnosis and tumour staging in patients with non-Hodgkin's lymphoma. These tumours are often multicentric and involve sites such as the liver or the spleen which are not easily accessible to surgical biopsy. Particularly with splenic involvement, there is a diagnostic problem of morphologically distinguishing the lymphoma cells in an admixture of normal lymphocytes. Since most lymphomas in adults are of B-cell origin, we studied the diagnostic value of adding a surface immunoglobulin (sIg) light chain analysis to the cytological evaluation of FNAs. B-clonal excess was determined by flow cytometric analysis of the sIg light chain distribution and a monoclonal finding was considered diagnostic of lymphoma. In primary diagnostic procedures the light chain analysis established a diagnosis of lymphoma in 5/14 (36%) aspirates from patients with poorly differentiated tumours. Fine needle aspirates performed as part of staging procedures were morphologically normal or inconclusive in 19 cases; in seven of these (37%) lymphoma involvement was diagnosed by the light chain analysis. Diagnostic precision was enhanced by combining morphological and immunological evaluation of fine needles aspirates in patients with established or suspected non-Hodgkin's lymphoma.     

High-grade non-Hodgkin''s lymphoma complicating polypoid nodular lymphoid hyperplasia and multiple lymphomatous polyposis of the intestine

Three patients with multiple lymphoid polyps of the small intestine--two with nodular lymphoid hyperplasia and one with multiple lymphomatous polyposis--developed high-grade B-cell lymphomas. A literature search has revealed only 10 previous cases of nodular lymphoid hyperplasia complicated by lymphoma and none of an association between multiple lymphomatous polyposis and high-grade lymphoma.     

Follicular lymphoma mimicking marginal zone lymphoma in lymph node: a case report

Nodal follicular lymphoma (FL) is typically composed of follicular or nodular proliferation of small cleaved lymphoid cells, presumably derived from germinal center (GC) B cells. The hallmark of FL is t(14;18)(q32;q21) chromosomal translocation, which juxtaposes anti-apoptotic gene BCL2 to immunoglobulin heavy chain (IGH) promoter. Reflecting this background, FL cells are immunohistochemically positive for BCL2 as well as GC B cell markers CD10 and BCL6. It is known that low grade B-cell lymphomas, including FL, chronic lymphocytic leukemia/small lymphocytic lymphoma, and marginal zone lymphoma, are sometimes associated with marginal zone differentiation or plasmacytic differentiation. The marginal zone differentiation obscures the morphological differences among these, providing diagnostic challenges for histopathologists. In this paper, we present a case of FL, originally mimicking marginal zone lymphoma in the axillary lymph node. Subsequent bone marrow biopsy showed paratrabecular infiltration of small to medium-sized lymphoid cells. Immunohistochemical analysis of the bone marrow biopsy together with histopathology and flow cytometry of the axillary lymph node led to a final diagnosis of FL with marginal zone differentiation in the axillary lymph node and its bone marrow infiltration. Our case illustrates and reconfirms the importance of clinicopathological correlation which leads to a correct diagnosis.     

Correlation of morphology, immunophenotype, and flow cytometry with remission induction and survival in high grade non-Hodgkin's lymphoma

A series of cases of high grade non-Hodgkin's lymphomas has been studied by morphology (110 cases), immunocytochemistry (90 cases), using reagents reactive in fixed paraffin-embedded tissue, and flow cytometry (77 cases). B-cell tumours constituted 67.0 per cent of the total, T-cell tumours 22.0 per cent, and unclassified cases 8.8 per cent. Immunocytochemistry revealed two anaplastic carcinomas. Of the 77 cases studied by flow cytometry, 67.5 per cent were diploid and 32.5 per cent DNA aneuploid. T-cell tumours were more likely to be diploid than B-cell tumours, though the difference did not reach statistical significance. T-cell tumours had a significantly lower proliferative index (%S + G2) than B-cell tumours (P = 0.002). The overall remission induction rate was 68 per cent and actuarial 3-year survival 47 per cent. There was a trend for cases with %S + G2 less than 22 per cent to survive longer (P = 0.07). This trend became statistically significant when aneuploid cases were added to the high PI group (P = 0.04). No correlation was seen between morphological or immunophenotypic groups and remission induction rates or survival.     

Relevance of Ki-67 expression in the classification of non-Hodgkin''s lymphomas: a morphometric and double-immunostaining study

The proliferation of reactive and neoplastic cells was retrospectively assessed in 92 cases of non-Hodgkin's lymphoma by morphometry using a double-immunoenzymatic technique including surface markers and the monoclonal antibody Ki-67. The findings were compared with the histological diagnosis. The overall Ki-67 positivity is not always a good measure of the corresponding corrected values and therefore we recommend that a correction should be made for the total number of complementary lymphocytes in the tumour. Taking the macrophages and the Ki-67 positivity of the reactive cells into account does not generally add any information. There was no difference in reactive cell content between follicular (counted within follicles) and diffuse lymphomas within the tumour areas. The value of the group mean for low-grade follicular (nodular) lymphomas was significantly higher than that of diffuse low-grade lymphomas, but not significantly different from that of intermediate-grade lymphomas. High-grade lymphomas exhibited significantly greater Ki-67 positivity than those of intermediate grade. In 76% of the cases there was significant agreement between malignancy grade (low/intermediate malignant versus high malignant) at 45% corrected Ki-67 counts.     

Prognostic significance of activation and differentiation antigen expression in B-cell non-Hodgkin's lymphoma

Immunophenotyping shows heterogeneity of expression of activation and differentiation antigens in B-cell non-Hodgkin's lymphoma (NHL). To investigate whether antigen expression correlates with clinical behaviour we have studied the clinical presentation and follow-up of a series of 111 B-cell lymphomas previously phenotyped for a panel of antigens including CD groups 5, 9, 10, 21, 23, 25, 30, 38, 4F2 antigen, and transferrin receptor. CD antigens 5, 10, and 23 were expressed significantly more often by low grade lymphomas whereas CD38, 4F2 antigen, and transferrin receptor were more often expressed by high grade lymphomas. There was a significant correlation with survival and age, stage at presentation, histological grade, and expression of 4F2 antigen and transferrin receptor but not with the other antigens studied. 4F2 antigen and transferrin receptor may identify a poor prognostic group of cases in low grade lymphoma but we conclude that phenotyping B-cell NHL for many of the antigens expressed at various stages of B-cell differentiation and activation does not provide clinically useful information in addition to that obtained from standard histological classifications.     

Primary non-Hodgkin's lymphoma of the intestine: a morphological, immunohistochemical and clinical study of 31 Chinese cases

Thirty-one cases of primary non-Hodgkin's lymphoma of the intestine were investigated. Twenty-one were of B-cell and 10 of T-cell origin. The B-cell lymphomas comprised two cases of low-grade B-cell lymphoma of mucosaassociated lymphoid tissue (MALT), one of centroblastic/centrocytic type, three of high-grade B-cell lymphoma coexisting with a low-grade B-cell lymphoma of MALT, nine of centroblastic, three of immunoblastic and three of Burkitt type. Of the T-cell lymphomas, eight were of pleomorphic medium-to large-sized cell type and two of large cell anaplastic type. All the B-cell lymphomas expressed CD20 (L26) and/or Ki-B5; in six there was monotypic immunoglobulin light chain restriction. Membrane positivity for CD45RO (UCHL1) was observed in the 10 cases of T-cell lymphoma, but the tumour cells did not express monocyte-macrophage markers. Clinically, the patients with T-cell lymphomas were usually young males with constitutional symptoms and their prognosis was significantly worse than those of patients with intestinal B-cell lymphoma.     

A quantitative exploration of surface antigen expression in common B-cell malignancies using flow cytometry

The use of flow cytometry to diagnose hematological malignancies has become routine due to its ability to often differentiate between morphologically similar diseases based on antigens expressed on the surface of malignant cells. In an attempt to expand on the utility of flow cytometry in the study of B-cell malignancies we have used the most reliable quantitative methodology, QIFI (quantitative indirect immunoflourescence assay), to study the expression of CD5, CD10, CD11c, CD19, CD20, CD22, CD23, and CD79b in 384 cases of several common B-lineage malignancies, including: B-ALL, CLL, SLL, hairy cell leukemia, diffuse large B-cell lymphoma, and follicular lymphoma. The impetus behind this extensive, single institution study of surface antigens was two-fold: evaluating similarities and differences of antigen expression between B-cell neoplasms and finding additional clinical utility for the quantitative flow cytometric data generated. Our results show that each distinct malignant histology has its own quantitative pattern of surface antigen expression. In most cases, these quantitative patterns do not increase the ability of flow cytometry to distinguish between them. However, a high expression of specific antigens on a given B-cell malignancy may potentially identify optimal therapeutic targets for current and/or future monoclonal antibody-based therapies.     

间变性大细胞淋巴瘤的p80蛋白表达及其临床意义

目的 研究染色体易位ｔ（２;５）所形成的嵌合基因ＮＰＭ－ＡＬＫ的蛋白产物－－Ｐ８０蛋白在间变性大细胞淋巴瘤（ＡＬＣＬ）中的表达及与其亚型和预后的关系。方法 对已进行临床、病理和免疫学分析胼有随访资料的１９例ＡＬＣＬ用免疫组织化学ＡＢＣ法标记Ｐ８０蛋白。结果 Ｐ８０蛋白在１９例中有９例呈强阳性,组织学亚型为普通型和小细胞型,无１例为霍金样型（Ｐ〈０．０５）。Ｐ８０蛋白阳性病例的免疫学表型为Ｔ细胞性产     

Ploidy, proliferative activity, p53 and bcl-2 expression in bronchopulmonary carcinoids: relationship with prognosis.

Bronchopulmonary well-differentiated neuroendocrine carcinoma (WDNEC) represents a more aggressive neoplasm than does typical carcinoid. Its biological behavior is variable and cannot be predicted on the basis of histopathological features. Nineteen typical carcinoids and 23 WDNECs were studied in order to obtain multiple parameters that should be used in the differential diagnosis between these two lesions and as prognostic markers of WDNEC. Flow-cytometry was performed on paraffin-embedded sections. Mutant p53 protein, the bcl-2 oncoprotein and the Ki-67 antigen were detected by immunohistochemical methods and evaluated quantitatively. WDNEC was more frequently aneuploid than typical carcinoid, had a higher percentage of Ki-67 positive nuclei and presented more frequently the mutant p53 protein. In WDNEC, the mutant p53 (p = 0.001), the bcl-2 oncoprotein (p = 0.002) and the high expression (> or = 16%) of Ki-67 (p = 0.0021) were associated with poor prognosis. The prognostic significance of mutant p53 and bcl-2 oncoprotein could be confirmed by Cox multiple regression survival analysis (p = 0.0005). It seems to be advisable to evaluate these features for the management of the patients affected by WDNEC.     

Defects in Thymocyte Differentiation and Thymocyte- Stromal Interactions in the Trisomy 16 Mouse

We have examined fetal thymic development in the trisomy 16 (Ts16) mouse, which is considered to be a model for human trisomy 21, or Down Syndrome. The Ts16 thymus contains 10 to 20% of the number of lymphocytes found in a normal thymus at a comparable stage. Expression of thymocyte differentiation markers (Thy-1, CD5, CD8, CD4, CD3, and HSA) is severely affected in Ts16 fetuses aged 14–18 gestational days. When thymuses from 14-day Ts16 mice were cultured in vitro, these markers eventually reached levels of expression comparable to those seen in normal thymuses in culture. On the other hand, expression of CD44 appears to be unaffected in Ts16 thymuses in vivo, but declines in vitro relative to normal thymuses. Reconstitution of depleted thymic stroma with thymocytes showed evidence of defects in both developmental compartments.     

Upregulation of nucleobindin expression in human-activated lymphocytes and non-Hodgkin's lymphoma

Nucleobindin (Nuc) was originally found to be an enhancement factor of antl-DNA antibody production secreted by a lymphold cell line derived from a lymphoprollferatlve MRL/Ipr mouse. It has been shown that Nuc has a unique structure containing a DNA- and two calcium-binding domains, and a leuclne zipper motif, but Its biological roles have not yet been fully elucidated. Expression of Nuc was first studied in human lymphocytes. Expression of Nuc mRNA in normal peripheral blood mononuclear cells was significantly increased upon mitogen stimulation. Anti-human Nuc monoclonal antibody H-1D8 immunopreclpitated Nuc protein in the nuclear extract of Molt-4 cells. Furthermore, in the immunohlstochemlcaf staining of tumor specimens from 108 patients with non-Hodgkin's lymphoma (NHL) with H-1D8, H-1D8-posltlve cells were observed in nearly all cases in varying frequency. According to the Working Formulation, the percentage of cases in which more than 90% of the tumor cells were stained with H-1D8 was 65% in the high grade of the histological malignancy, 54% In the Intermediate grade, and 22% in the low grade; however, normal ceils surrounding the tumor cells were virtually negative for H-1D8. These results showed that the level of Nuc expression in human lymphocytes reflects the status of activation or proliferation of the cells, thus providing a clue for the further investigation into biological roles of Nuc. In addition, It might be applicable to the clinlcopathological estimation of NHL as a novel Indicator.     

Cutaneous T cell lymphoma: Immunocytochemical study on activation/proliferation and differentiation associated antigens in lymph nodes,skin, and peripheral bloody

Summary The expression of activation/proliferation antigens (CD 25, CD 30, Ki 67, transferrin receptor) in lymph nodes and skin were compared in nine patients with mycosis fungoides (MF) with patients with erythroderma not related to MF, and patients with reactive lymphofollicular hyperplasias (a total of 14 patients). A panel of differentiation antigens was analyzed in addition. Reactivities were revealed by the APAAP technique. Activation/proliferation antigen scores in lymph nodes were related to the clinical stages of MF in most instances (low scores in cases of MF stage I/II, high scores in 3/4 cases of MF stage III/IV). They differed markedly in cases of non-MF-erythrodermia with the exception of one patient, and in all cases of reactive lymphofollicular hyperplasia. Expression of activation/proliferation antigens in lymph nodes were different in most cases from those in skin and peripheral blood. For diagnostic use, the activation/proliferation antigen scores were superior to the cell differentiation antigen profiles. Among cellular differentiation antigens, only the extent of CD1 + cells provided some diagnostic information, since the number of these cells were markedly increased in all cases of dermatopathic lymphadenitis with/without MF when compared with reactive lymphofollicular hyperplasia. In the diagnosis of MF, immunohistochemistry of activation/proliferation or differentiation antigens cannot replace routine paraffin histology, but may provide supplement any data in equivocal cases.Dedicated to Professor Dr. Theodor Nasemann on the occasion of his 65th birthday     

K562细胞株侧群细胞中部分白细胞分化抗原的表达

背景：白血病侧群细胞表型的研究对于理解肿瘤细胞的异质性和起源、分子标记和靶向治疗等都有积极意义。 目的：鉴定人慢性粒细胞白血病细胞株K562中是否存在侧群细胞,并观察侧群细胞亚群与非侧群细胞亚群中部分白细胞分化抗原的表达差异。 方法：采用流式细胞术检测K562细胞株中是否存在侧群细胞;并进一步分析K562侧群细胞和非侧群细胞两亚群间CD34+、CD34+CD38-、CD34+CD38+、HLA-DR+细胞的表达情况。 结果与结论：经Hoechst33342染色,流式细胞仪分析结果显示在K562中存在侧群细胞,这部分细胞比例少,为(2.7±0.5)%;统计学分析侧群细胞和非侧群细胞亚群中CD34⁺、CD34⁺CD38^-细胞表达率差异有显著性意义,而CD34⁺CD38⁺细胞表达率和HLA-DR+细胞表达率差异均无显著性意义;侧群细胞和非侧群细胞相比在分化抗原表达上有异质性。中国组织工程研究杂志出版内容重点：干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程全文链接：     

髓样细胞核分化抗原在强直性脊柱炎病人的表达

目的 通过比较强直性脊柱炎(AS)患者和健康志愿者的基因表达谱，寻找与AS相关的新基因并探讨其在病因和发病机制中的意义。方法 用含588个基因的cDNA微阵列，检测AS和健康志愿者的外周血单个核细胞的基因表达语，初步筛选出在AS中差异表达的基因，为了进一步验证cDNA微阵列差异表达基因结果，扩大各组病例数，再用RT-PCR技术同时检测、对比健康志愿者和AS患者外周血单个核细胞(PBMC)、关节液单个核细胞(SFMC)和关节滑膜细胞这些差异表达基因的变化。结果 和健康志愿者组的PBMC比较，AS病人骨髓样细胞核分化抗原(MNDA)、迁移抑制因子相关蛋白8(MRP8)和MRP14、粘附分子ICAM-1和integrin β1表达明显增高，其中MNDA和MRP8的变化相关系数为0．793。在AS的SFMC和关节滑膜细胞中，MNDA表达也显著增高(与健康志愿者组PBMC比较，P值分别为0．038和0．027)；MNDA区别AS和健康志愿者的统计学鉴别准确率达1。AS病人在接受抗TNF-α单克隆抗体治疗3个月后，MNDA在关节滑膜细胞的表达水平显著下降(P＝0．018)。结论 MNDA是AS发病过程中一个重要的并与单核／巨噬细胞致炎密切相关的免疫因子，其可能成为一种用于AS的诊断、治疗监控指标。     

Fine-needle aspiration cytology and flow cytometric immunophenotyping in diagnosis and classification of non-Hodgkin lymphoma in comparison to histopathology

This prospective study aimed to compare the value of fine needle aspiration (FNA) cytology (FNAC) and flow cytometric immunophenotyping (FCI) with histopatopathology (HP) in the diagnosis and classification of non-Hodgkin lymphoma (NHL). Twenty-nine excised lymph nodes suspected of NHL were evaluated using FNAC, FCI, and HP. Specimens were divided into two equal parts; one for HP and the other for FNAC and FCI. Results were compared in terms of diagnosis (malignant, benign or reactive, and metastatic) and NHL class. With combined FNAC/FCI, 11 (37.9%) cases were diagnosed as NHL, 11 cases (37.9%) as reactive lymph node, six cases (20.6%) as Hodgkin's lymphoma, and one case (3.4%) as metastasis. HP revealed nine cases (31%) of NHL, five cases (17.2%) of reactive lymph nodes and all the diagnosed metastatic and Hodgkin's lymphoma. Considering histology as a gold standard method in diagnosis, the sensitivity, specificity, PPV and NPV of FNAC/FCI in differentiate malignant and benign lesion were 73.9%, 83.3%, 94.4%, and 45.5%, respectively and in differentiate NHL from others were 75%, 93.8%, 90%, and 83.3%, respectively. Cytology and HP in addition to FCI and HP are significantly different from determination of NHL lesions point of view (P = 0.001 and P < 0.0001, respectively). However, FCI can be considered as an adjunctive method for Cytology especially because Cytology is not competent enough to differentiate between benign lesions and Lymphoma. Additionally, FCI is shown to be an accurate method in classifying NHL.