表型少见的慢性T淋巴细胞增殖性疾病2例报告

慢性T-淋巴细胞增殖性疾病在临床实践中较少遇见，其中的少见表型和变异型更是罕见。现报告1例CD4＋CD8-的大颗粒淋巴细胞白血病，1例由T慢性淋巴细胞白血病／小淋巴细胞性淋巴瘤转化的T-幼淋白血病，结合文献就其诊治进行讨论，以提高临床医生对该类疾病的了解和认识。     

10例慢性淋巴细胞白血病免疫表型分析

目的：探讨慢性淋巴细胞白血病（CLL）的免疫分型。方法：应用流式细胞仪对10例CLL患者外周血进行淋巴细胞表面抗原检测。结果：10例CLL患者CD19抗原阳性表达率为100%。其中1例患者CD5、CD19、CD20、HLA-DR、CD3、CD13均阳性,且CD34表达率大于10%;另有1例为CD5、CD19、CD20、HLA-DR阳性且伴CD7阳性。CD5、CD19、CD20、HLA-DR同时阳性的患者为50%。结论：CLL以B细胞慢淋为主,免疫分型对慢淋的诊断及鉴别诊断具有重要意义,其免疫分型可存在伴系表达。     

幼稚淋巴细胞白血病

幼稚淋巴细胞白血病(以下简称幼淋)是界于慢性淋巴细胞白血病(简称慢淋)与急性淋巴肉瘤细胞白血病(LcL)之间的一种类型。过去将此型归之于慢性LcL。多数认为是慢淋的变型,或为LcL的特殊类型。提高幼淋与慢淋及LcL的鉴别诊断,对决定治疗甚为重要。诊断: 幼淋的主要临床表现为:白细胞明显升     

慢性淋巴样白血病的研究进展(文献综述)

慢性淋巴样白血病是由不同疾患所组成的一类恶性疾病,其中大多数患者都为B细胞型,故又称“B细胞型慢性淋巴样白血病”。临床上主要包括三个疾病,即慢性淋巴细胞性白血病(CLL),幼淋巴细胞白血病(PLL)和毛细胞性白血病(HLC)。三者具有不同形态、临床和免疫特征,但均好发于老年患者。统计文献中211例HCL,确诊时平均年龄为52岁,最高达89岁,男女之比约3.6:     

单克隆B淋巴细胞增多伴肾功能不全

57岁男性,因血压升高伴肾功能减退2年入院,病程中体重减轻5 kg,脾大,外周血淋巴细胞增多,CD20~+ 1 420个/μl。骨髓流式细胞检测:淋巴细胞占有核细胞36.5%,其中B细胞占淋巴细胞78%,表达HLA-DR、CD5、CD19、CD20、CD22、sKappa(dim),lammda阴性。肾活检提示肾小球膜增生性病变伴单一IgG3沉积、肾间质多灶性CD20~+细胞浸润,最终诊断为膜增生性肾小球肾炎伴单克隆IgG沉积、B细胞慢性淋巴增殖性疾病-不能分类,予以利妥昔单抗治疗,随访13个月,血清肌酐、尿蛋白下降。对于病因不明的增生性肾炎伴持续性淋巴细胞增多者,需排除B细胞慢性淋巴增殖性疾病。     

老年急性淋巴细胞白血病患者免疫表型差异

目的探讨老年急性淋巴细胞白血病患者的免疫表型。方法选择2011年1月至2013年11月来该院接受治疗的55例老年急性白血病患者,取患者肘静脉血标本,分离外周血单核细胞并用流式细胞术对B细胞和T细胞进行免疫分型,评价不同免疫分型特征。结果 55例老年急性白血病患者中有35例(63.64%)B型淋巴细胞白血病,9例(16.36%)T型淋巴细胞白血病,11例(20.00%)B/T混合型淋巴细胞白血病,其中B型比例高于T型。B型淋巴细胞分化抗原表达异常患者中有33例(71.74%)CD19阳性,18例(39.13%)CD79a阳性,2例(4.35%)CD20阳性和3例(6.52%)CD10阳性。T型淋巴细胞分化抗原表达异常患者中有5例(25.00%)Cy CD3阳性,19例(95.00%)CD2阳性,18例(90.00%)CD4阳性,2例(10.00%)CD7阳性,2例(10.00%)CD5阳性。结论老年急性白血病患者主要以B淋巴细胞类型为主,且主要以CD19和CD79a为主。     

急性淋巴——粒细胞混合型白血病(附3例报告)

本所发现急性淋巴一粒细胞混合型白血病3例,现报告如下。例1,男6岁,患儿于1987年7月因反复发热3月,进行性苍白2月就诊,血象Hb57克/L,WBC6.5×10~9/L,plat232×10~9/L,分类早幼粒7%,幼淋巴19%。骨髓象(26/7):增生明显活跃,原始粒18.5%,早幼粒18%,淋巴系原始淋巴10%及幼稚淋巴细胞28%。细胞组织化学:POX阳性28%,PAS阳性76％,积分164,部分细胞呈粗颗粒状阳性反应。诊断为急性淋巴一粒细胞混合型白血病。取骨髓标本作电镜检查,细胞亚微结构形态学表现证实骨髓中存在着原始、幼稚淋巴及原始粒细胞.淋巴细胞免疫分析:E花环50%,M花环17%。混合花结:T54%,B15%,N30%,D1%。     

103例慢性淋巴细胞白血病患者免疫表型的研究

目的:探讨慢性淋巴细胞白血病(CLL)的免疫表型特征。方法:采用CD45/SSC双参数散点图设门,应用三色流式细胞术对103例初诊CLL患者骨髓或外周血标本进行免疫分型,另外选择同期5例T淋巴细胞增殖性疾病患者及92例急性淋巴细胞白血病(ALL),其中仅有的5例成熟急性淋巴细胞白血病(Mature-B-ALL)患者作为对照。结果:①103例患者中HLA-DR、CD19、CD20及CD5表达最常见,阳性率依次为100.0%、99.0%、96.0%及78.6%,T相关抗原CD2、CD3、CD4、CD7、CD8及CD25的表达率为3.0%、3.0%、1.9%、2.9%、1.0%和18.6%;5例T淋巴细胞增殖性疾病患者的HLA-DR、CD2和CD5表达阳性率最高,依次为100.0%、80.0%、80.0%,不表达B系相关抗原;CLL及T淋巴细胞增殖性疾病患者无一例表达CD1a、CD14、CD34及CD57;②CLL和T淋巴细胞增殖性疾病患者均伴有髓系抗原表达,CLL中CD13和CD11b的阳性率分别为51.1%和42.9%;T淋巴细胞增殖性疾病患者中CD13和CD11b的阳性率为100.0%和66.7%;③与ALL...     

胞浆内抗原CD_3、CD_(22)、MPO的检测方法及临床意义

目的 :探讨和明确胞浆内抗原 CD3、CD2 2 、MPO的检测方法及其在白血病诊断中的意义。方法 :采用多聚甲醛和甲醇联合固定细胞 ,单抗间接荧光法标记细胞内相应的抗原 ,在荧光显微镜下检测计数 ,并将结果与形态学和细胞表面抗原检测结果进行对照研究。结果 :19例初诊患者中 ,11例 MPO阳性 ,6例 Cy CD2 2 阳性 ,1例Cy CD3阳性 ,1例 3项均为阴性 ,结合形态学和表面抗原分析 ,全部病例获得明确诊断。结论 :胞浆内 CD3、CD2 2 、MPO的检测简便、准确、灵敏度高 ,可在临床广泛开展使用 ,对白血病的诊断和鉴别诊断具有重要意义     

微分化型急性髓系白血病5例临床分析

目的：探讨流式细胞免疫分型在微分化型急性髓系白血病（AML-M0）诊断中的作用。方法：分析5例AML-M0患者的临床资料,观察其骨髓细胞形态学和细胞化学染色的特征以及与流式细胞仪骨髓细胞免疫分型的符合程度。结果：5例患者骨髓细胞学均报告原始及幼稚淋巴细胞增多,POX染色阴性,PAS染色部分阳性,流式细胞免疫分型髓系标志CD13、CD33、CD117至少1项阳性,而淋巴系统及巨核细胞系统抗原阴性。结论：骨髓细胞形态学、细胞化学染色结合骨髓细胞免疫表型和细胞遗传学检查是诊断AML-M0的主要依据,其中原始细胞的免疫分型对诊断AML-M0是非常必要的。     

Quantitative analysis of CD79b, CD5 and CD19 in mature B-cell lymphoproliferative disorders.

BACKGROUND AND OBJECTIVE: Distinction between B-cell chronic leukemias can be difficult due to overlap in cell morphology and immunologic features. We investigated, by quantitative flow cytometry, the expression of CD79b, CD5 and CD19 in cells from a variety of B-cell disorders to see whether this analysis adds further information useful to the diagnosis and characterization of these diseases. DESIGN AND METHODS: Peripheral blood cells from 6 normal individuals were used as reference controls. The diseases of the 63 patients investigated comprised: 29 chronic lymphocytic leukemia (CLL), six of them with atypical morphology, 6 B-cell prolymphocytic leukemia (PLL), 12 splenic lymphoma with villous lymphocytes (SLVL) and 16 mantle-cell (Mc) lymphoma in leukemic phase. The study was carried out by triple immunostaining with directly conjugated monoclonal antibodies (MoAb) against CD79b, CD5 and CD19 and quantitative estimation of the antigens per cell assessed with standard microbeads (Quantum Simply Cellular). RESULTS: Compared to normal B-cells, the number of CD19 molecules was significantly lower in cells from all of the B-cell disorders except PLL. The intensity of CD5 in leukemic B-cells was significantly higher in CLL cells, including atypical cases, and Mc lymphoma than in normal B-cells, whilst PLL and SLVL had values similar to those of normal B-lymphocytes. CD79b was expressed at lower levels in all types of leukemic cells compared to normal B-lymphocytes but differences were statistically significant in CLL, Mc lymphoma and SLVL. The number of CD79b molecules per cell was significantly lower in typical CLL than in the remaining B-cell diseases whilst the comparison of CD5 and CD19 intensity between CLL and non-CLL samples failed to show any statistically significant difference. INTERPRETATION AND CONCLUSIONS: Distinct antigen density patterns for the various conditions emerged from this analysis: Typical CLL was characterized by moderate CD5 and weak or negative CD79b expression. Mc lymphoma showed an homogeneous pattern, characterized by similar expression of CD5 than CLL but significantly stronger expression of CD79b whilst PLL and SLVL had weak CD5 and moderate CD79b expression. Atypical CLL had an intermediate pattern of CD79b antigen expression ranging from weak to moderate with bright CD5. Unlike CD5 and CD79b, CD19 did not discriminate the various B-cell disorders but only between normal and leukemic cells.     

CD11c (LEU-M5) expression characterizes a B-cell chronic lymphoproliferative disorder with features of both chronic lymphocytic leukemia and hairy cell leukemia.

Chronic lymphocytic leukemia (CLL) and hairy cell leukemia (HCL) are two common chronic lymphoproliferative disorders, each having characteristic clinical, morphologic, and immunologic features. Phenotypically, CD5 reactivity in CLL and CD11c (Leu-M5) reactivity in HCL have characterized these two leukemias among B-cell disorders. In this study, we report 14 cases of a novel chronic lymphoproliferative disorder characterized by lymphocytosis and CD11c expression, but morphologically similar to CLL. The patients' ages ranged from 46 to 81 years (median 62). Eleven had palpable splenomegaly, five with markedly enlarged spleens; only one patient had generalized lymphadenopathy. The white blood cell count ranged from 5.2 to 131.0 x 10(9)/L (median 20.8). The morphologic diagnosis in all cases was CLL, with the cells usually having abundant cytoplasm. No morphologic features, of hairy cells were evident; tartrate-resistant acid phosphatase cytochemistry was negative in all cases. Bone marrow biopsies were available in 8 of 14. Four showed focal nodular infiltrates and two had diffuse infiltrates similar to CLL; two showed only minimal interstitial involvement. All cases expressed multiple B-cell markers, and 12 of 14 had monoclonal surface immunoglobulin. The leukemic cells of all cases strongly expressed CD11c, while CD5 was expressed in 7 of 14; only 1 of the 14 cases expressed the lymph node homing receptor, Leu-8. This unique group of leukemias appears to represent the malignant transformation of lymphocytes arising from a stage of lymphocyte differentiation between that found in typical cases of CLL and that of HCL. CD11c is known to have an important function in cellular adhesion and may be important in determining the pattern of lymphocyte tissue distribution found in this group of patients.     

In non-follicular lymphoproliferative disorders, IGH/BCL2-fusion is not restricted to chronic lymphocytic leukaemia

The translocation t(14;18) and its t(2;18) and t(18,22) variants, which involve the BCL2 genetic hallmark for follicular lymphoma (FL), have been reported in several cases of chronic B-cell lymphoproliferative disease (CLPD) and frequently in chronic lymphocytic leukaemia (CLL). We describe here the clinical, morphological, immunological, cytogenetic and molecular findings from 37 cases of t(14;18)-positive CLPD, identified from our series of non-FL B-cell neoplasms (n=993) that were routinely analysed in peripheral blood by conventional cytogenetics analyses. The FL diagnosis was excluded by morphology and immunology (the samples were CD10 negative in all cases). The BCL2 translocations were observed in 22 CLL cases, including 7 monoclonal B-cell lymphocytosis (MBL) cases re-classified according to the new International Workshop on CLL criteria, six small lymphocytic lymphoma (SLL) cases, 1 splenic marginal zone lymphoma (SMZL) case and eight cases of unclassifiable CLPD with overlapping CLL/MZL features. In the CLL cases, the IGH/BCL2 fusion was remarkably associated with trisomy 12 (13/22) and mutated IGHV status (20/21) and did not affect the outcome. Moreover, most of these CLLs harboured a low mutation load of BCL6 gene and unmutated FAS (CD95) loci, which points to a post-germinal-centre cellular origin.     

Characteristics of CD11c+CD5+ chronic B-cell leukemias and the identification of novel peripheral blood B-cell subsets with chronic lymphoid leukemia immunophenotypes.

Previous studies have indicated that chronic lymphocytic leukemias (CLL) are characterized by the coexpression of CD5 and B-cell antigens, while hairy cell leukemias (HCL) typically express CD11c+CD5- B-cell immunophenotypes. In this report we describe the features of B-cell leukemias with CD11c+CD5+ immunophenotypes and the identification of novel circulating B-cell subsets defined by the expression of CD20, CD5, and CD11c antigens. Morphologic evaluation of 14CD11c+CD5+ B-cell leukemias showed that they generally had larger cellular diameters (14 to 21 microns) and lower nuclear:cytoplasm ratios than typical small lymphocyte CLL. These cases did not exhibit the well-defined nucleoli characteristic of prolymphocytic leukemia (PLL). The presenting clinical features of CD11c+CD5+ B-cell leukemias were most consistent with CLL or PLL, and none of the evaluated cases had pancytopenia, splenomegaly, and cytoplasmic villi characteristic of HCL. Examination of normal peripheral blood (n = 6) by three-color flow cytometry identified four novel B-cell subsets with the following immunophenotypes (mean percent of total CD20+ B cells +/- SE): CD20+CD5+CD11c+ (8.0 +/- 1.6); CD20+CD5-CD11c+ (12.0 +/- 2.0); CD20+CD5+CD11c- (35.0 +/- 4.9); and CD20+CD5-CD11c- (44.0 +/- 5.0). Our findings suggest that CD11c+CD5+ B-cell leukemias with atypical morphologic features represent forms of CLL or PLL rather than HCL. In addition, we have identified novel subsets of circulating B cells defined by patterns of CD20, CD5, and CD11c expression that correspond to the immunophenotypes of chronic B-cell leukemias.     

Atypical chronic lymphocytic leukaemia witht(ll;14)(ql3;q32): karyotype evolution and prolymphocytic transformation

Summary. In order to define better the cytological and clinical features of atypical B-cell chronic lymphocytic leukaemia (B-CLL) with t(ll;14)(ql3;q32), sequential morphologic immunological and cytogenetic studies were performed in seven patients belonging to a series of 72 consecutive cases presenting with a diagnosis of CLL or atypical CLL according to the FAB criteria. Cytologic diagnosis in these seven patients with t(ll;14) was typical CLL in two cases presenting with < 10% large lymphocytes (LL) and prolymphocytes (PL) and atypical CLL in five cases in which LL and PL comprised between 10% and 55%. The diagnosis was supported by histologic findings on bone marrow biopsy (five cases) or splenectomy specimens (two cases). A progressive increase of peripheral LL and PL was observed, resulting in a switch of FAB diagnosis over a 6-60-month period from typical CLL into atypical CLL in two cases and from atypical CLL into prolymphocytic leukaemia in five cases. Immunophenotyping showed a mature B-cell phenotype with CD19, CD22, CD24 positivity and CD10 negativity in all patients. A bright-staining pattern for surface immunoglobulins (SIg) was detected in 6/7 cases, CD5 positivity in 6/7 cases, and CD23 positivity in 1/7 cases. The FMC-7 monoclonal antibody was positive in >40% cells in 5/6 cases. Chromosome changes in addition to t(11; 14) were seen in five cases; in two cases unbalanced translocations involving the 3q21 chromosome region, resulting in partial trisomy for the long arm of chromosome 3, were detected early in the course of the disease. Karyotype evolution that was associated with disease progression occurred in 3/6 assessable patients. Comparison of these findings with similar data from 65 B-CLL patients without t(ll;14) showed that atypical morphology, switch of FAB diagnosis during the course of the disease, and karyotype evolution were more frequently seen in cases with t(ll;14) (5/7 v 15/65 cases, P = (V015, 7/7 v 7/65 cases, P < 0-0001, and 3/6 v 5/45 assessable cases, P= 0-04, respectively). The frequency of positivity for CD2 3 and bright SIg staining differed significantly in the two groups. It is concluded that t(ll;14) identifies a cytologically atypical subset of B-CLL, characterized by frequent cytologic and cytogenetic evolution and by a distinct immunological profile, sharing some biological features with mantle cell lymphoma.     

CD5-Negative,CD10-Negative small B-cell leukemia: Variant of chronic lymphocytic leukemia or a distinct entity?

CD5- and CD10-negative chronic lymphocytic leukemias are quite uncommon as compared to the CD5-positive CLL. We reviewed 250 sequential cases of peripheral blood lymphocytosis to characterize cases of small B-cell lymphoproliferative disorders, submitted with a clinical diagnosis of chronic lymphocytic leukemia exhibiting a non-classic immunophenotypic profile. Six cases of CD5-, CD10-negative chronic lymphocytic leukemias and no tissue involvement were identified that revealed high-density surface-membrane immunoglobulin and CD20 expression, with variable expression of CD11c, CD23, and CD25. Most had a profound leukocytosis (mean WBC 180 x 10(9)/L) with proliferation of mature-appearing lymphocytes. Subsequent bone marrow biopsies showed diffuse infiltration by neoplastic cells in all evaluated patients. The clinical course appeared indolent, with follow-up revealing three patients alive (survival time 38-68 months), while two died of unrelated causes and one was lost to follow-up soon after diagnosis. These cases may represent somewhat unusual chronic lymphoproliferative disorders, with morphologic features and immunophenotypic profile not readily classifiable, but which are certainly atypical for classic chronic lymphocytic leukemia. Some of these features are reminiscent of those seen in marginal-zone lymphoma. However, it is most unusual for this known to be tissue-based disease to present primarily as leukemia rather than lymphoma.     

Distinctive features of "nurselike" cells that differentiate in the context of chronic lymphocytic leukemia.

A subset of blood mononuclear cells from patients with chronic lymphocytic leukemia (CLL) can differentiate in vitro into "nurselike" cells (NLCs) that can protect CLL cells from apoptosis. NLCs express cytoplasmic vimentin and stromal-derived factor 1 (SDF-1). NLCs also express CD14, as well as CD11b, CD33, CD40, CD45RO, CD68, CD80, CD86, HLA-DQ, and HLA-DR, but not CD1a, CD2, CD3, CD11c, CD19, CD45RA, CD83, CD106, or CD154. Consistent with this phenotype, NLCs failed to differentiate from blood mononuclear cells that were depleted of CD14+ cells or from isolated CD19+ cells. CD14+ blood cells of healthy donors could differentiate into cells with the morphology and phenotype of NLCs when cultured in direct contact with CLL B cells, but not with normal B cells. Despite expressing antigens in common with blood monocytes, monocyte-derived dendritic cells, and macrophages, NLCs expressed significantly higher levels of CD68 than these other cell types. Consistent with the notion that NLCs are present in vivo, CD14+ splenocytes from CLL patients have NLC morphology and express significantly higher levels of CD68 than CD14+ splenocytes from persons without known B-cell malignancy. These findings indicate that although NLCs may differentiate from blood monocytes, they probably represent a distinctive hematopoietic cell type that exists in vivo, differentiates from hematopoietic CD14+ cells in the context of CLL, and in turn protect CLL cells from apoptosis via a mechanism that is independent of CD106 (vascular cell adhesion molecule-1). The interaction between CLL cells and NLCs may represent a novel target for therapy of patients with this disease.     

Distinct morphophenotypic features of chronic B-cell leukaemias identified with CD1c and CD23 antibodies.

Morphological criteria usually applied to diagnose various subtypes of B-cell chronic lymphoid leukaemia are largely subjective. Immunophenotyping of 61 relevant cases using a selected panel of monoclonal antibodies (mAb), showed that CD1c and CD23 mAb were able to separate B-cell chronic lymphocytic leukaemia (B-CLL) from other chronic B-cell lymphoproliferative diseases. Lymphocytes of B-CLL were CD1c-, CD23+, whereas those of other types of chronic B-cell leukaemia were CD1c+/-, CD23-, and CD38/-. Non-B-CLL cases had a significantly higher amount of large peroxidase-negative (unstained) cells analyzed with an automated blood cell counter (Technicon H6000). This type of volumetric assessment allowed a separation between typical and "atypical" B-CLL, which otherwise were both CD1c-, and CD23+. These combinations of phenotypic markers corresponded to well-defined haematopathologic entities, conventionally diagnosed on peripheral blood (PB) and bone marrow smears, and on histologic sections of lymph nodes and spleen.     

Possible specific chromosome change in prolymphocytic leukemia

The chromosomes of unstimulated and stimulated blood lymphocytes from 5 cases with B-cell prolymphocytic leukemia (PLL) were examined following the use of polyclonal B-cell activators (PBA). Banding techniques revealed a common and specific chromosome abnormality to be present in each of the cases, which was due to a translocation between chromosomes 6 and 12 (t(6;12)(q15;p13]. The fact that this specific chromosome change has not been reported in other lymphoproliferative disorders may indicate that PLL is a distinct clinical entity and different from other lymphoproliferative disorders, whether it occurs de novo or complicates chronic lymphocytic leukemia (CLL).     

Prolymphocytic leukaemia: surface morphology in 21 cases as seen by scanning electron microscopy and comparison with B-type CLL and CLL in ''prolymphocytoid'' transformation

The surface architecture of leukaemic cells obtained from 21 cases of proven prolymphocytic leukaemia (PLL) and eight cases of chronic lymphocytic leukaemia (CLL) with 'prolymphocytoid' transformation (PL-CLL) was compared with the cell surface morphology of leukaemic cells obtained from 46 cases of B-type CLL, using the scanning electron microscope (SEM). All cases were defined by cytochemistry, immunological markers and transmission electron microscopy prior to SEM examination. B-CLL cells showed the well-recognized spectrum of surface architecture described in earlier studies. The majority of cells had moderate numbers of short microvilli, although in a minority, cells with relatively smooth surfaces predominated. In seven of the eight cases of PL-CLL, cells were villous in nature and in this respect similar to CLL cells; however, more cells with dense microvilli were seen. The prolymphocytic cells were recognized by their larger size and in 18 of the 19 cases of B-derived PLL, villous cells predominated. Two cases of T-derived PLL showed variable cell surface morphology ranging from smooth to moderately villous. It appears that B-PLL cells are most frequently villous and display more surface microvilli than B-CLL cells. B-prolymphocytes display the surface features regarded as characteristic for neoplastic B-cells as seen in patients with B-type lymphoma and leukaemia.