急性淋巴细胞性白血病细胞及其骨髓基质细胞粘附分子表达和粘附行为的探讨

分析了５８例急性淋巴细胞性白血病（ＡＬＬ）患者和１０例正常人骨髓单个核细胞及其骨髓基质细胞上粘附分子的表达。发现与正常人相比，白血病细胞上表达的ＣＤ１１ａ、ＣＤ１１ｂ、ＣＤ１８、ＣＤ３４、ＣＤ４４、ＣＤ４９ｄ和基质细胞上表达的ＣＤ５４，ＣＤ４９ｂ均有显著性改变（明显下降或显著增高）；同时，来自ＡＬＬ的基质细胞在粘附分子表达和粘附行为上亦不同于正常骨髓基质细胞，认为粘附分子的异常表达可能参与了ＡＬＬ的发病机制。     

PCR-SSCP分析急性淋巴细胞白血病TCR基因寡/亚克隆重排及临床意义

目的：为研究急性淋巴细胞白血病（ALL）TCR基因寡／亚克隆重排及与临床关系。方法：采用PCR－SSCP技术分析81例ALL TCR VγI-Jγ基因重排。结果：65例（80．2％）ALL病人存在TCR VγI-Jγ基因重排,8例（8．6％）病人存在寡／亚克隆重排;寡／亚克隆ALL的白细胞计数显著高于非寡／亚克隆ALL（P＜0．01）;寡／亚克隆ALL的完全缓解率为37．5％,显著低于非寡／亚克隆ALL（80．7％）（P＜0．005）。结论：应用PCR－SSCP方法可确定ALL的寡／亚克隆性,ALL病人寡／亚克隆检测有助于预测疗效和预后判断。     

急性淋巴细胞白血病患者HLA-DRB1基因多态性研究

目的 :研究北方汉族人群HLA DRB1等位基因多态性与急性淋巴细胞性白血病 (ALL)患者的遗传关联。方法 :采用聚合酶链反应 序列特异性寡核苷酸探针 (PCR SSO)方法 ,对 184例北方汉族人群ALL患者和 3 5 78例健康脐带血样本HLA DRB1等位基因多态性进行研究。结果 :184例ALL患者HLA DR9( 18 2 1%,χ2 =16 .0 7,P <0 0 1,RR =1 74,EF =0 0 774) ,DR12 ( 14 13%,χ2 =7.12 ,P <0 0 1,RR =1 5 2 3,EF =0 0 49) ,DR14( 7 88%,χ2 =5 .194,P <0 0 5 ,RR =1 5 74,EF =0 0 2 87)和DR16 ( 4 89%,χ2 =8.5 2 7,P <0 0 1,RR =2 0 6 5 ,EF =0 0 2 5 )基因频率显著高于正常人群 ,而HLA DR7( 8 15 %,χ2 =7.0 78,P <0 0 1,RR =0 6 0 ,EF =0 0 86 )和DR15 ( 12 2 3%,χ2 =4.0 49,P <0 0 5 ,RR =0 710 9,EF =0 0 6 7)基因频率显著下降。结论 :HLA DR9、DR12、DR14、DR16等位基因对ALL患者有遗传易感作用 ,而HLA DR7、DR15等位基因对ALL患者有遗传拮抗作用。     

急性淋巴细胞白血病患者BAALC基因表达水平也许有临床意义

目的研究急性淋巴细胞白血病（ALL）患者脑和急性白血病细胞胞浆（BAALC）基因的表达水平及其临床意义。方法57例经细胞形态学、组织化学染色及流式细胞术免疫学分型确诊的ALL患者,其中男性35例,女性22例;中位年龄23（14～66）岁。FAB分型L19例。L245例,L33例。建立实时荧光定量PCR检测BAALC基因表达的技术,并定量检测57例初发ALL患者BAALC基因的表达水平及进一步分析其与临床疗效的关系。结果57例初发ALL患者BAALC基因表达水平显著高于对照组。FAB各亚型中BAALC基因表达水平差异无统计学意义（P〉0．05）,BAALC基因表达水平的高低与初发AI上患者的免疫表型具有显著性相关．T—ALL患者和伴有髓系抗原表达ALL患者BAALC基因表达水平显著高于B—ALL（P〈0．05）和不伴有髓系抗原表达ALL患者（P〈0．01）。ALL中BAALC基因表达的水平与外周血白细胞计数（WBC）、血红蛋白（Hb）、血小板计数（Plt）及骨髓白血病细胞比例无明显相关性（P〉0．05）。BAALC基因高表达ALL患者的完全缓解率（73．3％）同低表达组f81．0％）比较差异无统计学意义（P〉0．05）,但BAALC基因高表达患者1年复发率（81．8％）明显高于低表达组（44．1％）（P〈0．05）。结论BAALC基因高表达提示可能是ALL患者一个不良预后因素,检测ALL患者BAALC基因表达水平可能有助于指导治疗。     

急性淋巴细胞性白血病细胞及其骨髓基质细胞粘附分子表达和…

分析了５８例急性淋巴细胞性白血病患者和１０例正常人骨髓单个核细胞及其骨髓其质细胞上粘附分子的表达。发现与正常人相比，白血病细胞上表达的ＣＤ１１ａ，ＣＤ１１ｂ，ＣＤ１８，ＣＤ３４，ＣＤ４４，ＣＤ４９ｄ和基质细胞上表达的ＣＤ５４，ＣＤ４９ｂ均有显著性改变。     

急性淋巴细胞白血病患者GLUT3、TSLC1的表达及临床意义

目的:探讨急性淋巴细胞白血病患者葡萄糖转运蛋白-3(Glucose transporter 3,GLUT3)、肿瘤抑制物1(Tumor suppressor in lung cancer 1,TSLC1)的表达及临床意义.方法:选取2018年4月至2020年4月期间我院诊治的59例急性淋巴细胞白血病患者作为研究对象的研究组,另选取53例同期非急性淋巴细胞白血病患者作为研究对象的对照组.比较两组患者骨髓单个核细胞中GLUT3、TSLC1的表达情况,以及患者的不同临床病理特征对骨髓单个核细胞中GLUT3、TSLC1表达的影响.结果:研究组GLUT3阳性表达率高于对照组、TSLC1阳性表达率低于对照组(P<0.05);血红蛋白(异常)、白细胞计数(异常)者GLUT3阳性率明显高于血红蛋白(正常)、白细胞计数(正常)者(P<0.05).白细胞计数(异常)者TSLC1阳性率明显高于白细胞计数(正常)者(P<0.05).结论:在急性淋巴细胞白血病患者骨髓单个核细胞中GLUT3阳性表达率高、TSLC1阴性表达率高.     

CD4+CD25+Treg在急性淋巴细胞白血病患者外周血中的表达及其体外实验研究

目的初步探讨CD4 CD25 调节性T细胞(CD4 CD25 regulatory T cells,CD4 CD25 Treg)在急性淋巴细胞白血病(acute lymphocytic leukemia,ALL)患者化疗前及化疗缓解后外周血中的表达水平,并研究患者血清能否诱导外周血CD4 CD25-T细胞转化为CD4 CD25 Treg。方法①采用流式细胞术分别检测ALL初诊组、化疗完全缓解或部分缓解组及正常对照组外周血中CD4 CD25 T细胞所占比例,然后通过荧光定量RT-PCR检测各组外周血中转录因子Foxp3mRNA的表达水平,并逐层分析比较。②采集正常人外周血单个核细胞后,对照组用正常人血清,实验组用患者血清并分别设浓度梯度进行培养,72h后采用流式细胞术、荧光定量RT-PCR分别检测CD4 CD25 T细胞和Foxp3mRNA表达。结果ALL化疗缓解组CD4 CD25 T细胞及Foxp3mRNA表达水平均明显高于ALL初诊组和正常对照组(P<0.05),后两者之间CD4 CD25 T细胞水平无统计学差异(P>0.05),但ALL初诊组Foxp3mRNA含量较正常对照组明显升高(P<0.01),差异具有统计学意义;并且血清培养对照组CD4 CD25 T细胞水平及Foxp3mRNA含量均明显低于实验组(P<0.05),且其表达并不随血清浓度的增加而升高。结论CD4 CD25 Foxp3 Treg在ALL初诊组及化疗缓解组患者外周血中比例明显升高,且初步表明患者血清中的可溶性物质可诱导外周血CD4 CD25 T细胞转化为CD4 CD25 Treg,提示CD4 CD25 Treg可能是ALL免疫抑制的一个重要原因。     

急性淋巴细胞白血病浸润牙龈1例

患者男性 ,30岁 ,在无任何诱因下出现左上颌牙龈包块2个月。检查 :贫血貌 ,颌下可触及肿大淋巴结 ,5 6 78颊侧面有一 5ｃｍ× 4ｃｍ× 2 5ｃｍ大小的肿块 ,表面呈菜花状 ,有糜烂。实验室检查 :Ｈｂ 6 6ｇ/Ｌ ,ＷＢＣ 2 4× 10 9/Ｌ ,Ｐｌｔ 190×10 9/Ｌ ,Ｌ 0 37,Ｎ 0 6 0 ,骨髓象增生明显活跃 ,淋巴系统显著增生 ,其中原幼淋占 0 6 5。临床初步诊断 :左上颌牙龈白血病浸润。遂活检左上颌牙龈包块以进一步证实。病理检查　灰白组织一块 2ｃｍ× 1 5ｃｍ× 1ｃｍ ,切面灰白 ,质细嫩 ,呈鱼肉状。镜检 :牙龈黏膜下方为弥漫浸润的肿瘤性原…     

细胞凋亡与急性淋巴细胞白血病治疗的研究进展

近年来,由于对细胞凋亡研究的深入,急性淋巴细胞白血病的临床治疗也有了新的可能。通过选择性诱导白血病干细胞的凋亡,可对急性淋巴细胞白血病的治疗开辟新的方向。而观察病程中细胞凋亡和其相关因子的情况,也可对急性淋巴细胞白血病的预后起到预报作用。虽然此类治疗方法并未大批量应用于临床,但相信随着细胞凋亡机理的进一步研究和其他工具学科的发展,细胞凋亡的选择性诱导也终将成为治疗急性淋巴细胞白血病的有效途径。因此,本文将对细胞凋亡和急性淋巴细胞白血病治疗的研究进展做一综述。     

急性白血病患者骨髓细胞中TIMP3、RUNX3蛋白表达情况检测及其临床意义

目的： 检测急性白血病(AL)患者骨髓细胞中的TIMP3、RUNX3蛋白表达的情况及其与基因甲基化的关系,并探讨其与AL患者预后的关系。方法: 采用Western blotting检测50例AL患者骨髓单个核细胞(BMMCs)及10例健康供者外周血单个核细胞(PBMCs)中TIMP3、RUNX3蛋白表达的情况,结合前期实验中TIMP3、RUNX3启动子区域甲基化检测结果,对与AL患者预后有关的因素进行统计分析。结果: 50例AL患者BMMCs中RUNX3甲基化组该蛋白表达明显少于未甲基化组及正常对照组(P<0.05),AL患者完全缓解(CR)率与RUNX3蛋白表达及骨髓中原始细胞比例有关,RUNX3蛋白失表达、骨髓中原始细胞比例高的患者CR率低,反之较高。结论: RUNX3启动子区域甲基化是导致该蛋白失表达的原因,其参与了AL的发病,并与不良预后有关,TIMP3甲基化与急性白血病发病无关。     

A direct bone marrow chromosome technique for acute lymphoblastic leukemia

We describe a direct bone marrow chromosome technique that was developed especially for use in studies of acute lymphoblastic leukemia (ALL). The features responsible for technical improvements include: the use of RPMI 1640 medium, supplemented with 30% fetal calf serum, to support cellular activity during both specimen transport and Colcemid treatment; the processing of only 0.1 ml of sedimented cells or less per centrifuge tube; the exposure of cells to Colcemid for a maximum of 25 min; control of the total time of exposure to hypotonic solution; the use of a steel wire as a stirring rod (fashioned to fit the centrifuge tube) for mixing cells; slide preparation by a specific edging-flaming technique; the natural aging of the slides to achieve optimal drying; and the use of a modified G-banding procedure that employs Wright's stain. This technique has been used in more than 350 cases of ALL and has consistently provided analyzable banded chromosomes, even in hyperdiploid cases with up to 91 chromosomes. It makes the previously recognized morphological difference between metaphases of residual normal cells and those from the leukemic clone less apparent. The edging-flaming technique of slide preparation is the most important component and is especially appropriate for spreading large numbers of chromosomes in ALL.     

Detection of residual acute lymphoblastic leukemia cells in cultures of bone marrow obtained during remission

We used a semisolid culture assay to quantitate leukemia cells in the bone marrow of patients with childhood acute lymphoblastic leukemia (ALL). In bone marrow cultures from 40 patients with newly diagnosed disease, the colonies that developed in vitro consisted of lymphoblasts with the same surface markers and abnormal karyotype as the original diagnostic marrow specimens. We also studied marrow cultures from 13 patients in chemotherapy-induced remission; 6 of these, including 1 obtained from a patient during successful engraftment after marrow transplantation, also yielded lymphoblast colonies in culture, with the same immunologic phenotype or abnormal karyotype as the original leukemic marrow. Four of these patients, including the one who underwent marrow transplantation, relapsed within 2 to 30 months of the abnormal cultures; the other two are still in remission, one of them 30 months after diagnosis. Bone marrow cultures from eight normal controls and from the other seven patients in remission did not yield lymphoblast colonies; all seven of the latter are still in remission. This assay appears to allow detection of small numbers of residual leukemic cells. We conclude that the technique will be valuable in monitoring the efficacy of chemotherapy and allogeneic bone marrow transplantation in acute lymphoblastic leukemia, as well as in evaluating the quality of purged marrow for autologous marrow transplantation.     

干扰素-α联合GM－CSF诱导慢性髓性白血病细胞向树突状细胞分化

目的：研究干扰素-α（IFN-α）对慢性髓性白血病（CML）骨髓单个核细胞来源的树突状细胞（DCs）发育的影响。 方法： 12例初发慢性期CML患者的骨髓单个核细胞，分别与含如下细胞因子,RPMI-1640培养液共育：rhGM-CSF 1×106U/L联合rhIFN-α 2×106U/L（IFN-α组）、rhGM-CSF 1×10⁶U/L联合rhIL-4 5×10⁵U/L（IL-4组）、单用rhGM-CSF 1×10⁶U/L和单用rhIFN-α 2×106U/L，培养7 d；于第8-10 d，部分孔加入rhTNF-α 5×10⁴U/L。形态学（Wright染色、倒置显微镜）、免疫学（CD80、CD86、CD83、CD1a、HLA-DR）检测；磷脂酰丝氨酸（PS）转位检测 DCs凋亡情况；荧光原位杂交（FISH）对1例CML进行细胞遗传学分析；混合淋巴细胞反应（MLR）检测刺激同种异体T淋巴细胞增殖的能力。 结果： CML骨髓单个核细胞经上述细胞因子诱导7 d后，IFN-α组和IL-4组均呈现树突状细胞的典型形态；免疫学鉴定，IFN-α组DCs CD80、CD86、CD83、HLA-DR的表达显著高于IL-4组（P<0.05），经5×104U/L rhTNF-α作用后，两组DCs CD80、CD86、CD83、HLA-DR进一步上调，其中IFN-α组DCs CD80、CD86、CD83、HLA-DR的表达显著高于IL-4组（P<0.05）；经FISH证实来源于白血病细胞；两组DCs均具有刺激同种异体T淋巴细胞增殖的能力，IFN-α组刺激淋巴细胞增殖的能力明显高于IL-4组(P<0.05)。 结论： IFN-α可促进CML骨髓单个核细胞来源的树突状细胞的分化、活化。这可能是IFN-α在CML中的治疗机制之一。     

急性淋巴细胞白血病患儿骨髓基质细胞的粘附行为

目的和方法:用MTT方法检测了急性淋巴细胞白血病(ALL)患儿骨髓基质细胞(BMSC)对正常骨髓造血细胞和淋巴瘤Raji细胞的粘附行为,用流式细胞仪检测了BMSC表面的粘附分子ICAM-1和VCAM-1的表达。结果:急性期组ALLBMSC对骨髓造血细胞的粘附能力明显下降。长期缓解组ALLBMSC对肿瘤细胞的粘附能力明显增高。急性期组ALLBMSC表面ICAM-1表达明显降低。结论:ALL患儿BMSC对骨髓造血细胞和肿瘤细胞的粘附行为的异常,BMSC粘附行为的变化与细胞表面粘附分子的表达异常有关。     

Immunosuppressive properties of mesenchymal stem cells derived from bone marrow of patients with chronic myeloid leukemia

Mesenchymal stem cells (MSCs) have emerged as excellent candidates for clinical application because of their capabilities of differentiating into multiple mesenchymal lineages and supporting hematopoiesis. Recently, MSCs have gained further interests after the demonstration of an immunosuppressive role. However, it is still unclear whether the immunosuppressive capability of MSCs will be altered with disease state. In this study, we obtained and expanded MSCs from bone marrow of patients with chronic myeloid leukemia (CML). Our results showed that MSCs derived from CML do not express costimulatory molecules CD40, CD80, and CD86. When MSCs derived from CML were added back to T cells stimulated by mitogens, a significant inhibition of T-cell proliferation was evident. MSCs differentiated into various mesenchymal lineages did not alter their immunosuppressive effect on T-cell proliferation. A significant T-cell inhibition was found in a transwell system, in which cell-cell contact between MSCs and effector cells was prevented. Furthermore, we found that transforming growth factor beta1 (TGF beta1) and hepatocyte growth factor (HGF) were major mediators of T-cell suppression by MSCs derived from CML. These results demonstrated that autologous MSCs derived from CML could effectively suppress T-cell proliferation.     

Immunophenotypic analysis of acute lymphoblastic leukemia using routinely processed bone marrow specimens

Monoclonal antibodies have been recently developed that react with antigens expressed on T and B lymphocytes in routinely processed, paraffin-embedded lymphoid tissues. In this study, we assessed bone marrow clot and/or core biopsy sections of 19 cases of acute lymphoblastic leukemia (ALL) using routinely decalcified, B5- or formalin-fixed, paraffin-embedded sections and a panel of monoclonal antibodies, including LN1, LN2, L26, Leu-22, UCHL-1, and LCA. Each case had been previously phenotyped using freshly obtained aspirate material and a standard immunophenotypic protocol. Our results demonstrate the utility of the LN2 antibody in differentiating between precursor B-cell (pre-B) and precursor T-cell ALL. The LN2 antibody stained 11 of 12 cases of pre-B ALL and did not react with any of the seven T-cell ALLs. The other antibodies tested were less helpful. The Leu-22 antibody stained both pre-B and T-cell ALLs, while the results with UCHL-1 revealed peculiar nuclear staining of pre-B and T-cell ALLs; this we attributed to processing artifact. The L26 antibody reacted with only one case of pre-B ALL (also CD20 antigen positive), while the LN1 antibody did not react with any pre-B ALLs. Neither L26 nor LN1 stained any cases of T-cell ALL. The LCA antibody stained in only four (21%) of 19 cases, two pre-B and two T-cell ALLs. The results also suggest that this panel of antibodies may be useful in differentiating ALL from mature B-cell and T-cell lymphomas involving the bone marrow.     

Spontaneous death of bone marrow and peripheral blood cells during remission of acute lymphoblastic leukemia in children

Programmed antileukemic chemotherapy is associated with prolonged cytotoxic effects on not only tumor cells, but on intact bone marrow and peripheral blood cells as well. The maximum spontaneous cell death is observed immediately after the end of therapy. It decreases with time after therapy, but does not reach the level observed in healthy subjects and does not depend on the type of therapy. The percentage of dead cells in peripheral blood directly correlates with the percentage of dead cells in the bone marrow. The processes of cell death in the peripheral blood and bone marrow are synchronous and parallel.