恶性淋巴瘤骨髓受累的免疫表型特征

目的:探讨淋巴瘤骨髓受累的免疫表型特征。方法:采用流式细胞仪CD45/SSC设门方法对34例恶性淋巴瘤患者的骨髓标本进行检测,以骨髓涂片细胞学检查作阳性对照。收集骨髓受累患者的CD分子表达数据。结果:①对34例恶性淋巴瘤患者的骨髓应用流式细胞仪进行检测,发现23例阳性,阳性率67.65%(23/34),95%可信区间(51.92%,83.37%)。②该23例阳性患者中,非霍奇金淋巴瘤(NHL)19例,霍奇金淋巴瘤(HL)4例。NHL患者中B细胞来源免疫荧光单克隆抗体标记抗原出现频率最高的为CD19,CD20;T细胞来源标记抗原出现频率最高的为CD7。而在HL患者中出现频率最高的为CD9。结论:采用流式细胞仪CD45/SSC设门方法,发现非霍奇金淋巴瘤骨髓受累患者免疫表型特征为:B细胞来源:CD19、CD20;T细胞来源:CD7。霍奇金淋巴瘤为:CD9。     

Immunophenotype of c-kit cells in normal human bone marrow: implications for the detection of minimal residual disease in AML

Summary. In the present study, seven normal human bone marrow samples from healthy volunteers have been analysed in order to investigate the immunophenotypic characteristics of the normal CD117⁺ cells and their utility for the detection of minimal residual disease in 71 acute myeloid leukaemia patients.
Our results show that most of normal BM CD117⁺ cells coexpress the HLADR and the myeloid associated CD33 antigen. In addition, almost half of CD117⁺ cells are CD34⁺, these cells displaying a different FSC/SSC distribution when compared to the CD117⁺/CD34⁻ cells. No CD117⁺/CD15⁺ and CD117⁺/CD10⁺ cells were detected and very few CD117⁺ cells (<1 × 10⁻³) expressing the HLADR⁻/CD34⁻, CD33⁺/HLADR⁻ and CD34⁺/HLADR⁻ phenotypes were found to be present in normal BM. In contrast, from the 71 AML patients analysed, 34 had CD117⁺/CD15⁺ blast cells and eight had the CD117⁺ phenotypes detected at low frequencies (<1 × 10⁻³) in normal BM.
In summary, the present study shows that the use of the CD117 antigen in different monoclonal antibodies combinations may be of great help for the detection of minimal residual disease in a high proportion of AML cases, especially in those patients displaying the CD117⁺/CD15⁺ phenotype, because cells coexpressing both antigens in normal BM, if present, are at very low frequencies.     

探讨Double/Triple Hit淋巴瘤的流式免疫表型特征在疾病诊断及生存结局相关性中的应用

目的：本研究目的是确立二次或三次打击淋巴瘤(Double/Triple- Hit lymphoma,D/THL)流式免疫表型特征,研究与B淋巴细胞白血病(B-LBL)在免疫表型上的差异,并探讨流式免疫表型特征与D/THL生存结局的相关性。方法 采用多色FCI分析法对40例D/THL患者和25 B-LBL患者的样本的免疫表型进行检测和评价,并将流式免疫表型结果与基因亚型以及总体生存率进行统计学对比分析。结果 90%(36 / 40)的D/THL病例CD10表达呈阳性,近一半病例(48%,19 / 40)缺乏表面轻链。45%(18 / 40)的病例CD45表达呈弱阳性, 40%(16 /40)CD19表达呈弱阳性, 43%(17 / 40)CD20表达呈弱阳性。28%(11 / 40)显示伴随CD19和CD20的弱表达,38%(15 / 40)的CD19和CD20表达呈差异化表达(variable expression)。在不同基因亚型之间,除Dim CD45和 BCL2阳性发生率达有差异外(BCL2+/MYC+组Dim CD45发生率明显低于BCL2+/BCL6+/MYC+组(p=0.03);BCL2+/MYC+组中bcl2阳性率为96%,而其它基因亚型组阳性率均为0%),其它表型发生率差异均无统计学意义。与B-LBL相比,B-LBL的CD45和CD20低表达发生率明显高于D / THL(c2 = 20.78、20.83,);CD19和CD20同时弱表达在D / THL发生率稍多(28%D/ THL vs. 10% B-LBL),但差异无统计学意义(c2 = 2.31)。B-LBL的TdT阳性发生率明显高于D / THL(cc2= 30.74,)。对30例已经死亡患者进行了生存分析显示,不同基因型总体生存率无显著性差异(BCL2+ MYC+对BCL6+/MYC+,P=0.08;BCL2+MYC+对BCL2+/BCL6+/MYC+,P=0.13;BCL6+/MYC+对BCL2+/BCL6+/MYC+,P=1.0);Dim CD45和Bcl2阳性预示着与较低的总体生存率(P = 0.01,0.015)相关。 结论D / THL流式免疫表型特征研究有助于D / THL鉴别诊断。对流式免疫表型与疾病结局的生存分析结果,将有利于临床对D／THL整体预后进行危险分层管理,以便能针对特定病人及时采取个体化治疗。     

Treatment with short-term,high-dose cyclosporin A in children with refractory chronic idiopathic thrombocytopenic purpura

We report on 14 children (seven boys, seven girls) with chronic idiopathic thrombocytopenic purpura (ITP) refractory to multiple treatments, who were given a short-term therapy (range between 6 and 10 weeks) with high doses of cyclosporin A (CyA) (median, 10 mg/kg/d). Six patients experienced adverse events and one developed severe systemic mycosis during therapy. A complete response (CR) was observed in four patients and a partial response (PR) in three patients. Only the four CR patients, who were all girls, had a sustained response. These data suggest that CyA may be effective in some children with chronic symptomatic ITP.     

A marked decrease in l‐selectin expression by leucocytes in infants with Bordetella pertussis infection: leucocytosis explained?

OBJECTIVE: Infants with Bordetella pertussis infection (whooping cough) have an unexplained lymphocytosis and leucocytosis characterized by an increase in small lymphocytes with convoluted and cleaved nuclei. To characterize these cells immunophenotyping using multiparameter flow cytometry was performed on leucocytes from a group of 11 infants aged 3-6 months with proven pertussis and from uninfected control subjects. METHODOLOGY: The panel of monoclonal antibodies used to elucidate leucocyte subtypes included activation, adhesion, costimulatory, memory, T-helper (Th) 1 and Th2 markers. RESULTS: Patients with pertussis showed an increase in absolute numbers of neutrophils, monocytes, T lymphocytes (both CD4 and CD8), B lymphocytes (including CD10+/CD19+ haematogones) and natural killer (NK) cells. All leucocyte subgroups showed a marked decrease in L-selectin (CD62L) expression. The expression of other adhesion molecules CD11a, CD44 and CD54 on all leucocyte subgroups was unchanged. Expression of costimulatory molecules, CD49D and CD28 on T cells and CD80 and CD86 on monocytes, was unchanged. Lymphocyte activation markers CD69, CD25 and HLA-DR were unchanged. There was an increase in CD45RA+/CD45RO+/CD4+ cells (activated) and CD62L-/CD45RO+/CD4+ cells (Th1-like) but no increase in CD7-/CD4+ T cells (Th2-like). CONCLUSIONS: L-Selectin expression mediates extravasation of leucocytes into tissues and is important for homing of peripheral blood lymphocytes to lymph nodes. The significant down-regulation of L-selectin on leucocytes in pertussis infection may prevent leucocyte migration to areas of infection and homing and adhesion of T and B cells to peripheral lymphoid tissues. The increase in lymphocytes with Th1 phenotype may be required for effective immune response to the infective organism. These data provide a possible explanation for the absolute leucocytosis observed in this disease.     

Immune parameter analysis of children with sickle cell disease on hydroxycarbamide or chronic transfusion therapy

Sickle cell disease (SCD) is increasingly appreciated as an inflammatory condition associated with alterations in immune phenotype and function. In this cross‐sectional study we performed a multiparameter analysis of 18 immune markers in 114 paediatric SCD patients divided by treatment group [those receiving hydroxycrabamide (HC, previously termed hydroxyurea), chronic transfusion (CT), or no disease‐modifying therapy] and 29 age‐matched African American healthy controls. We found global elevation of most immune cell counts in SCD patients receiving no disease‐modifying therapy at steady state. Despite the decrease in percentage of haemoglobin S associated with CT therapy, the abnormal cellular immune phenotype persisted in patients on CT. In contrast, in both univariate and multivariate analysis, treatment with HC was associated with normalization of the vast majority of leucocyte populations. This study provides additional support for HC treatment in SCD, as it appears that HC decreases the abnormally elevated immune cell counts in patients with SCD.     

125例成人急性髓系白血病免疫表型特点分析

目的：探讨成人急性髓系白血病(acute myelogenous leukemia,AML)患者的免疫表型特点及临床意义。方法2011年5月-2014年6月在河北省涿州市医院和解放军总医院就诊的125例成人AML患者为研究对象,采用多色流式细胞术,以CD45/SSC双参数散点图设门,对其骨髓进行免疫表型检测。结果 AML患者髓系特异性抗原表达阳性率依次为CD13(90.4%)、CD33(86.4%)、MPO(80.0%)、CD117(73.6%);CD14和CD15主要在M4和M5中表达,CD64和CD15同时强阳性伴HLA-DR高表达,提示M5可能性大;在M3患者中有较强的自发荧光,表达髓系抗原CD13、CD33和MPO,低表达CD34和HLA-DR。造血干/祖细胞标记抗原CD34、HLA-DR、CD38在AML中呈高表达,分别为53.6%、70.4%、71.2%。125例中56例伴有淋系抗原表达,各抗原表达率依次为CD56(25.6%)、CD7(19.2%)、CD19(9.6%)、CD4(7.2%)、CD2(6.4%)。有淋系抗原表达(LymAg+)组表达CD34+比例和肝脾大患者比例明显高于无淋系抗原表达(LymAg-)组(P＜0.01)。伴有CD7+患者完全缓解(CR)率明显低于CD7-患者(P＜0.05),而伴有淋系抗原表达的AML患者CR率(55.6%)与未伴有淋系抗原表达的AML患者CR率(76.2%)差异无统计学意义(P＞0.05)。结论 AML患者高表达CD13、CD33、MPO,部分患者同时表达髓系、淋系抗原;伴有CD7+表达的患者,化疗后CR率低,预后较差。     

基底细胞样乳腺癌的病理诊断

目的:探讨基底细胞样乳腺癌的病理形态特征,免疫表型、鉴别诊断及最新的研究进展。方法：观察20例基底细胞样乳腺癌的组织形态、免疫组化特点及临床资料,结合文献进行分析。结果：20例均为女性,年龄25-71,平均年龄45岁;镜下形态学特征主要包括推挤样边缘,地图样坏死,高级别核及核分裂易见。免疫组织化学结果：ER、PR、CerbB2均阴性表达,CK5/6有15例明显表达,占75%,5例弱表达,占25%,EGFR在7例病例中有表达,占35%。结论：基底细胞样乳腺癌是一种特殊类型的乳腺癌,具有独特的形态学特征及免疫表型,是一种独立亚型,应该提高对此类疾病的认识。     

Deciphering the genotype and phenotype of hairy cell leukemia: clues for diagnosis and treatment

ABSTRACT

Introduction: Hairy cell leukemia (HCL) is a rare, indolent B-cell neoplasm. The classical variant of the disease is characterized by the BRAF V600E mutation, which is present in virtually all cases. How this mutation leads to the signs and symptoms of the disease is currently not known.

Areas covered: This review explores the genetic background of HCL, especially the BRAF V600E driver mutation, but passenger mutations and their effects are also included. The clinical significance of BRAF mutations in other cancer types is discussed, as well as BRAF- induced senescence. An overview of the major forms of treatment of HCL (cytostatic drugs, specific BRAF inhibitors, B cell-specific antibodies) is given. Finally, possible mechanisms of the monocytopenia and hairy morphology so typical of this disease are discussed.

Expert opinion: Although being a rare disease, HCL and its pathogenesis can yield important information about BRAF-related cancer metabolism. Many aspects of the disease are still unclear, but with the right resources, this could change. This can lead to a more efficient and specific treatment, thus leading to decreased morbidity.