白细胞介素8及其受体在急性白血病的表达及其意义

目的探讨白细胞介素８（ＩＬ８）及其Ａ、Ｂ受体（ＩＬ８ＲＡ、ＩＬ８ＲＢ）的表达与急性白血病（ＡＬ）的分型、疗效及并发症的关系。方法用酶联免疫夹心法检测７７例初诊ＡＬ患者外周血ＩＬ８含量,用间接免疫荧光及流式细胞术检测骨髓单个核细胞（ＭＮＣｓ）表达ＩＬ８Ｒ,并检测了１５例ＡＬ完全缓解（ＣＲ）期脑脊液（ＣＳＦ）中ＩＬ８水平。结果ＡＬ患者外周血ＩＬ８水平高于正常人,其中急性髓系白血病（ＡＭＬ）高于急性淋巴细胞白血病（ＡＬＬ）,Ｍ４、Ｍ５高于Ｍ１～Ｍ３,ＢＡＬＬ高于ＴＡＬＬ（Ｐ＜０．０５）。ＡＬＬ中ＩＬ８＞１００ｎｇ／Ｌ组比≤１００ｎｇ／Ｌ组疗效差（Ｐ＜０．０５）;约３６．３６％的ＡＬ患者ＩＬ８Ｒ（＋）,ＩＬ８Ｒ（＋）者的外周血白细胞计数及幼稚细胞比例显著高于ＩＬ８Ｒ（－）者（Ｐ＜０．０５）;ＣＲ期患者ＣＳＦ中ＩＬ８水平与治疗前比较,差异无显著性（Ｐ＞０．０５）,合并中枢神经系统白血病（ＣＮＳＬ）时明显升高。结论外周血和ＣＳＦ中ＩＬ８水平的检测及检测骨髓ＭＮＣｓ膜ＩＬ８Ｒ的表达有助于ＡＬ各亚型的鉴别及预后判断;ＣＮＳＬ发生时ＣＳＦ中ＩＬ８增高。     

急性白血病骨髓细胞外基质的改变

作者应用组织化学、免疫组织化学染色及自动图象分析仪观测５９例急性白血病（ＡＬ）患者１１８次骨髓活检标本骨髓细胞外基质（ＥＣＭ）成分变化及其与造血细胞增殖的关系。其中男性３８例，女性２１例，中位数年龄３４岁。急性非淋巴细胞白血病（ＡＮＬＬ）４１例，急性淋巴细胞白血病（ＡＬＬ）１８例，初诊未治者（ＰＬ）２７例，难治复发者（ＲＬ）３２例，正常对照１０例。结果显示，ＡＬ与正常人骨髓ＥＣＭ成分有显著性差异（Ｐ＜０．０５）；ＡＬ网硬蛋白增加，纤维连接蛋白（Ｆｎ）、酸性粘多糖减低，中性粘多糖初诊未治者增高，难治复发者减低。ＡＬＬ和ＡＮＬＬ之间未见显著性差异；ＲＬ中难以缓解与复发再治者比较，后者改变更明显，而同复发次数无显著性差异。ＰＬ中追踪观察，６／８例（７５％）未缓解患者及８／１９例（４２．１％）完全缓解后复发患者骨髓基质Ｆｎ、酸性粘多糖、中性粘多糖有不同程度改变。骨髓基质Ｆｎ与造血细胞增殖在正常对照组显示呈正相关，而在ＡＬ呈负相关，ＰＬ与ＲＬ尚存在显著性差异（Ｐ＜０．０５）。提示ＡＬ骨髓造血微环境ＥＣＭ有明显变化，与发病机制及预后关系密切。     

急性白血病骨髓基质细胞肿瘤坏死因子α分泌活性及其意义

目的：探讨急性白血病（ＡＬ）骨髓基质细胞肿瘤坏死因子α（ＴＮＦα）分泌活性。方法：应用体外骨髓基质细胞培养法及ＴＮＦα生物活性检测法，对ＡＬ患者２０例及正常对照者１０名的骨髓基质细胞培养上清中的ＴＮＦα活性水平进行检测，并同步检测了患者血清、骨髓白血病细胞培养上清中的ＴＮＦα活性。结果：ＡＬ骨髓基质细胞形成能力较差，但其ＴＮＦα活性明显高于对照组（Ｐ＜０．０５）；少部分急性髓系白血病（ＡＭＬ）骨髓白血病细胞能自发分泌ＴＮＦα；患者血清中的ＴＮＦα活性显著高于正常对照（Ｐ＜０．００１）。结论：ＡＬ患者骨髓基质细胞存在量和质的异常，ＴＮＦα的异常分泌可能是导致ＡＬ增殖的一个重要因素     

白血病细胞膜相关因子（MAF－J6－1）单克隆抗体的制备及性质

用部分纯化的白血病细胞膜相关因子（ＭＡＦ－Ｊ６－１）对ＢＡＬＢ／Ｃ小鼠脾细胞进行体外免疫，然后与ＳＰ２／０小鼠骨髓瘤细胞融合获杂交瘤，经３次再克隆得到能产生ＭＡＦ－Ｊ６－１单克隆抗体（单抗）的杂交瘤细胞系。用ＡＢＣ免疫酶标法检测，该单抗在Ｊ６－１细胞上产生强阳性反应，并与ｒｈ－Ｍ－ＣＳＦ单抗对ＭＡＦ－Ｊ６－１及ｒｈ－Ｍ－ＣＳＦ产生交叉中和反应。ＭＡＦ－Ｊ６－１单抗还对Ｊ６－２、ＬＣＬ、Ｋ５６２等细胞系呈膜阳性反应，对白血病患者骨髓中部分原始和幼稚的单核、粒细胞呈阳性反应。在２４例被检测的白血病患者中，有三例呈强阳性反应，ＭＡＦ－Ｊ６－１可用作研究ｍ－Ｍ－ＣＳＦ与白血病关系的工具。     

30例急性白血病自体骨髓移植后造血功能重建的影响因素

对３０例急性白血病（ＡＬ）自体骨髓移植（ＡＢＭＴ）后造血功能恢复进行了单因素和多因素分析。移植后ＷＢＣ＞１．０×１０￣９／Ｌ、ＡＮＣ＞０．５×１０￣９／Ｌ、ＢＰＣ＞２０×１０￣９／Ｌ和胸骨髓增生活跃的中位时间分别为３０，３７，６１和５０天。白血病类型（急性髓细胞白血病）和骨髓细胞－１９６℃冻存是造血功能尤其是血小板延期恢复的主要因素（Ｐ＜０．０５），而年龄、性别、缓解（ＣＲ）至ＡＢＭＴ间期、停化疗至采髓及移植的间期、预处理方案及回输的骨髓细胞量等对造血功能的恢复无明显影响（Ｐ＞０．１）。血小板的延期恢复造成输血及血小板量明显增加和住院时间明显延长（Ｐ＜０．０１）。移植后９０天ＢＰＣ＜２０×１０￣９／Ｌ者早期复发的可能性较大（Ｐ＝０．０６）。     

血尿酸与代谢相关脂肪性肝病患者肝功能异常的相关性及因果研究

摘要：目的 研究血尿酸（ＵＡ）与代谢相关脂肪性肝病（ＭＡＦＬＤ）患者肝功能异常的相关性及因果关系。方法 纳入于四川大 学华西医院就诊的 ＭＡＦＬＤ 患者 ９８ 例。用 Ｓｐｅａｒｍａｎ 相关和多元 Ｌｏｇｉｓｔｉｃ 回归分析 ＵＡ 与丙氨酸氨基转移酶（ＡＬＴ）、天冬氨酸 氨基转移酶（ＡＳＴ）间的相关性。用双向孟德尔随机化研究分析 ＵＡ 与 ＡＬＴ、ＡＳＴ 之间的因果关系。结果 ＵＡ 与 ＡＬＴ、ＡＳＴ 呈 正相关，相关系数分别为 ０．５６（Ｐ＜０．００１）和 ０．４６（Ｐ ＝ ０．０１）。高 ＵＡ 组相较于低 ＵＡ 组 ＡＬＴ（ＯＲ ＝ ３．５８，Ｐ ＝ ０．００３）、ＡＳＴ（ＯＲ ＝ ３．２４，Ｐ＝ ０．００５）水平增加。正向因果中，ＵＡ 每升高 １ 标准差（ＳＤ），ＡＬＴ 增加 ０．０７ＳＤ（Ｐ ＝ ０．００２），ＡＳＴ 增加 ０．０６ＳＤ（Ｐ ＝ ０．０３）。 而在反向因果中，尚未发现 ＡＬＴ、ＡＳＴ 与 ＵＡ 间相关（Ｐ＞０．０５）。结论 ＵＡ 可导致 ＡＬＴ、ＡＳＴ 水平升高，是 ＭＡＦＬＤ 患者肝功能 受损潜在的危险因素。     

血清可溶性L—选择素,sICAM—1,HBV DNA拷贝数及肝功能损害的相关性研究

目的 探讨Ｌ－ｓｅｌｅｃｔｉｎ（Ｌ－选择素）和ＩＣＡＭ－１（细胞间粘附分子－１）在乙型肝炎病毒感染造成肝功能损害过程中的作用。方法 定量检测４２例乙型肝炎病人血清ＨＢＶ ＤＮＡ拷贝数、可溶性Ｌ－选择素（ｓＬ－ｓｅｌｅｃｔｉｎ）、可溶性细胞产粘附分子－１（ｓＩＣＡＭ－１）及肝功能相关生化指标,并对结果进行统计分析。结果 ２２例ＤＮＡ阴性和２０例ＤＮＡ阳性病人血清ｓＬ－ｓｅｌｅｃｔｉｎ和ｓＩＣＡＭ－１水平均明显高于对照组（Ｐ〈０．０５）,但ＤＮＡ阴性组和阳性组之间的ｓＬ－ｓｅｌｅｃｔｉｎ和ｓＩＣＡＭ－１水平均无显著差异（Ｐ〉０．０５）;ＤＮＡ拷贝数与ｓＩＣＡＭ－１水平呈负相关（ｒ＝－０．５０１,Ｐ〈０．０５）;ＤＮＡ拷贝数与肝功能损害呈负相关;４２例乙肝病人的ｓＬ－ｓｅｌｅｃｔｉｎ和ｓＩＣＡＭ－１水平均与肝功能损害     

MDM2和p53基因蛋白在急性白血病细胞中表达及与化疗反应的关系

目的：研究人急性白血病（ＡＬ）细胞ＭＤＭ２及ｐ５３基因蛋白表达水平、它们的相互关系及对ＡＬ化疗反应预测的价值。方法：免疫组化方法检测ＭＤＭ２及ｐ５３蛋白。结果：①４６例ＡＬ细胞ＭＤＭ２蛋白阳性率为７１．７％，ｐ５３蛋白阳性率为２１．７％，无细胞类型不同的差别（Ｐ＞０．０５），复发难治组比初发未治组约高１倍；②ＭＤＭ２与ｐ５３蛋白呈反向表达者多见，以ＭＤＭ２（＋），ｐ５３（－）方式表达（６７．４％）高于同向方式表达（１５．２％）（Ｐ＜０．０１）；③ＭＤＭ２蛋白阴性者骨髓缓解（ＢＭＲ）率（６９．２％）高于阳性者（３３３％）（Ｐ＜０．０５），两种蛋白不同表达关系对化疗效应有影响；④ＡＬ患者４例中２例化疗后ＭＤＭ２蛋白转阴性，获ＢＭＲ，另２例则反之。结论：人ＡＬ细胞ＭＤＭ２蛋白阳性率高，ｐ５３蛋白阳性率低；在同一ＡＬ细胞中两者多呈反向表达，且不同表达方式对化疗有影响；在ＡＬ细胞中确实存在ＭＤＭ２（＋），ｐ５３（－）表达方式。ＭＤＭ２蛋白，特别是ＭＤＭ２蛋白与ｐ５３蛋白联检可能预测ＡＬ细胞对化疗的敏感性     

急性白血病患儿外周血白血病细胞TfR表达与细胞增殖力和铁代谢的关系

目的探讨急性白血病患儿外周血白血病细胞转铁蛋白受体表达与细胞增殖力和铁代谢的关系。方法采用放射性配体结合分析法测定细胞转铁蛋白受体（ＴｆＲ）位点数及解离常数Ｋｄ值。结果和结论急性淋巴细胞白血病（ＡＬＬ）ＴｆＲ位点数为（７．８２６±６．０５４）×１０４／细胞,Ｋｄ为（６４６８±４．７７７）ｎｍｏｌ／Ｌ;急性髓系白血病（ＡＭＬ）ＴｆＲ为（２０．４０６±１７．８７６）×１０４／细胞,Ｋｄ为（８．６８３±４８９０）ｎｍｏｌ／Ｌ,两组间Ｋｄ值差异无显著性。完全缓解组ＴｆＲ位点数明显低于复发、死亡组。两组白血病细胞和经植物血凝素刺激的外周血单个核细胞ＴｆＲ位点均与细胞３ＨＴｄＲ掺入值呈正相关,说明ＴｆＲ表达与细胞增殖力有关。ＡＬＬ和ＡＭＬ组血清铁蛋白（ＳＦ）、细胞内铁蛋白（ＣＦ）均较对照组高（Ｐ＜０．０５）,Ｔｆ、铁结合力较对照组低,血清铁水平变化不大。ＴｆＲ位点与ＳＦ和ＣＦ均呈正相关,与Ｔｆ呈负相关。     

简便,灵敏的天冬氨酸转氨酶同工酶电泳法

以醋纤薄膜为载体，半胱氨酸亚磺酸为氨基供体，ｍ－ＰＭＳ和ＭＴＴ为显色剂，电泳分离天冬氨酸转氨酶同工酶ｍ－ＡＳＴ和Ｃ－ＡＳＴ。灵敏度达２．２Ｕ／Ｌ，批内重复性７．５Ｕ／Ｌ（ｎ＝１０）ＣＶ＝９．５％；２１．４Ｕ／Ｕ（ｎ＝１０）ＣＶ＝８．４％。用１４０Ｕ／Ｌｍ－ＡＳＴ作倍化稀释考核其线性，结果为Ｙ＝０．０８５２９＋０．９９６２Ｘ（理论值），ｒ＝０．９９９７。与Ｔｅｒａｎｉｓｈｉ法比较，Ｙ＝１．７５５８＋０．８３１１Ｘ（本法），ｒ＝０．８９４０。     

Roles for the heliodynamic hormones, all trans retinoic acid and 1 alpha, 25-dihydroxyvitamin D3, in control of the hematopoietic cell cycle

It is now well established that the production of primary hematopoietic cells is controlled at different levels of the biological organization. Bone marrow (BM) stromal cells, the extracellular matrix (ECM), polypeptide hematopoietic growth factors (HGF) as well as endogenous cell-division cycle (CDC) related factors play a dominant role in this control. Recent information suggest that the 2 lipophilic hormones, transRA and 1 alpha,25D3, depending on and/or perhaps mediating solar energy, play a role in the maintenance of BM homeostasis. Here we show that both transRA and 1 alpha,25D3: a) modulate the growth and/or stimulate the adipocytic differentiation of fibroblastic stromal cells (F-CFU); b) inhibit the synthesis and extracellular processing but stimulate the solubilization of matrix collagen; c) modulate the clonal growth of myeloid progenitor cells (GM-CFU) in synergy with HGFs; and d) inhibit the production of lactic acid in standard, normal long-term BM cultures (LTBMC). Comparative analysis of normal, preleukemic and leukemic BM cells in LTBMC indicated a positive correlation between the induction of terminal differentiation and reduced lactate production elicited by transRA or 1 alpha,25D3. These results raise a hypothesis according to which the terminal differentiation induced by the helicodynamic hormones is dependent on the mitochondrial aerobic ATP-generating system whose impairment may be a critical step during the process of leukemic transformation.     

Long-term marrow culture of cells from patients with acute myelogenous leukemia. Selection in favor of normal phenotypes in some but not all cases.

Long-term cultures were initiated with leukemic marrow aspirate cells from each of 13 newly diagnosed acute myelogenous leukemia (AML) patients. Initial assessment of the clonogenic potential of the marrow suggested that normal hemopoietic progenitors were reduced in most cases and progenitors of abnormal colonies and clusters were present in 10 cases. Subsequent assays of both nonadherent and adherent fractions of long-term cultures revealed two patterns of progenitor cell behavior. The most common pattern (nine cases) featured the detection after 1-4 wk of near normal numbers of typical erythroid, granulopoietic, and mixed colony-forming progenitor cells. Progenitors of abnormal (blast) colonies and clusters initially demonstrable in eight of these nine cases were, in these cases, not sustained in long-term culture and could not be found after 4 wk. Conversion to cytogenetic normalcy in long-term culture was confirmed in two experiments in this group. The second pattern (four cases) was characterized by the failure of progenitors capable of normal differentiation to become detectable in long-term cultures, and the concomitant maintenance of blast progenitors in the two cases in this group where such cells were initially demonstrable. Although progenitors capable of producing abnormal (blast) colonies or clusters in methylcellulose were not detected in either of the other two experiments, the maintenance for 6 wk of a hypercellular nonadherent blast population in one of these suggested the persisting activity of an "adherent layer-dependent" leukemic progenitor cell. Taken together, these findings indicate a strong correlation between the presence of leukemic blasts and their progenitors and a decreased level of normal hemopoiesis. In addition, the failure of leukemic cells to be maintained in long-term marrow cultures from some (but not all) AML patients suggests new applications of this methodology for studies of early stages of leukemic cell development.     

Membranes replace irradiated blast cells as growth requirement for leukemic blast progenitors in suspension culture

The blast cells of acute myeloblastic leukemia (AML) may be considered as a renewal population, maintained by blast stem cells capable of both self-renewal and the generation of progeny with reduced or absent proliferative potential. Blast progenitor renewal is manifested in suspension culture by an exponential increase in clonogenic cells. This growth requires that two conditions be met: first, the cultures must contain growth factors in media conditioned either by phytohemagglutinin (PHA)-stimulated mononuclear leukocytes (PHA-LCM), or by cells of the continuous bladder carcinoma line HTB9 (HTB9-CM). Second, the cell density must be maintained at 10(6) blasts/ml; this may be achieved by adding irradiated cells to smaller numbers of intact blasts. We are concerned with the mechanism of the feeding function. We present evidence that (a) cell-cell contact is required. (b) Blasts are heterogeneous in respect to their capacity to support growth. (c) Fractions containing membranes from blast cells will substitute for intact cells in promoting the generation of new blast progenitors in culture. (d) This membrane function may be specific for AML blasts, since membranes from blasts of lymphoblastic leukemia or normal marrow cells were inactive.     

HLA identical leukemia cells and T cell growth factor activate cytotoxic T cell recognition of minor locus histocompatibility antigens in vitro.

Lymphocytes from a healthy HLA-identical bone marrow transplant donor were tested for their ability to destroy her brother's acute myelogenous leukemia blasts in vitro. Primary mixed lymphocyte culture (MLC) and cell-mediated lysis (CML) responses between the patient's remission (pretransplant) and donor's lymphocytes were negative. Stimulation of donor lymphocytes for 7 d in vitro with irradiated leukemia cells, leukemia cells plus allogeneic irradiated lymphocytes, or a pool of irradiated lymphocytes from 10 donors, did not activate any cytotoxic cells able to destroy the HLA identical leukemic blasts. Further culturing for 7 additional d in T cell growth factor (TCGF) generated lymphocytes that induced effective cytotoxicity against the leukemic blasts, but not against autologous lymphocytes. Effective killing against the leukemia was observed only in cultures initially stimulated with the irradiated leukemia cells. These cytotoxic cells were maintained in TCGF and mediated persistent killing against the leukemic target cells. They were also able to destroy lymphocytes from the patient's mother and father, but not from an unrelated cell donor. This suggested specific recognition of non-HLA antigens inherited by the patient, that were foreign to the HLA identical bone marrow donor. These lymphocytes were cloned by a limiting dilution technique and one clone maintained cytotoxicity to the AML blasts and the father's lymphocytes, but not lymphocytes from the mother or an HLA-identical donor. This cytotoxicity was inhibited by a monoclonal anti-HLA antibody. Thus, in vitro sensitization of this sibling's lymphocytes with AML blasts followed by TCGF expansion, and cloning, enabled the detection of HLA-restricted cytotoxic cells that recognize minor locus histocompatibility antigens. This immune recognition may be relevant to the "graft vs. leukemia" effect that has been observed in leukemic animals and patients following histocompatible hematopoietic transplants.     

急性髓细胞白血病细胞来源的树突状细胞的诱导及功能研究

为了研究急性髓细胞白血病(AML)细胞诱导成树突状细胞(DC)的方法及其DC的功能，分离25例AML患者骨髓或外周血单个核细胞，在含有细胞因子rhGM—CSF，rhIL-4，rhTNF—α的IMDM完全培养基中培养8—12天，进行细胞形态学、表型、遗传学及功能测定。结果表明：20／25例自诱导培养的第3天起，在倒置显微镜下可观察到部分细胞体积增大，形态由圆形变得不规则，可见驼峰样或细刺状胞浆突起；在第8—9天，该类细胞所占的比例达到峰值。至第12天，细胞总数及上述形态的细胞数量明显减少。诱导培养结束时收集细胞，应用流式细胞术检测11／20例诱导前后细胞表面标志的表达，结果表明诱导前细胞不表达CD1a、CD80及CD83，低表达CD86、CD54和HLA—DR；诱导后细胞CD1a、CD80、CD83、CD86、CD54及HLA—DR的表达明显上调。在同种异体混合淋巴细胞反应(Allo-MLR)中，该类细胞刺激同种异体淋巴细胞增殖一对^3H-TdR的摄取能力明显高于AML细胞。FISH证实诱导生成的具有DC特征细胞的AML起源。结论：体外联合应用细胞因子可将AML细胞诱导成具有DC形态、表型及功能特征的细胞(AML—DC)。     

造血生长因子对急性髓系白血病细胞动力学耐药的克服

髓系白血病细胞“细胞动力学耐药”是指处于G_0期或增殖缓慢、细胞周期长于药物接触时间的白血病细胞对某些细胞周期特异性细胞毒药物产生耐药的现象。阿糖胞苷(Ara-C)为目前治疗急性髓系白血病(AML)标准诱导方案的药物之一。它是一种通过抑制DNA的合成,特异性地杀伤处于S期的白血病细胞的细胞周期特异性的细胞毒药物。~3H-胸腺嘧啶核苷体内标记研究证实,在AML存在一部分处于增殖静止状态和增殖缓慢的白血病细胞。体外研究发现,某些造血生长因子(HGF),如干细胞因子,IL-3,GM-CSF和G-CSF,在单用或联合应用条件下,不但能动员髓系白血病细胞进入细胞周期,使其对Ara-C的敏感性增加,细胞周期动力学耐药得以克服,而且对正常造血祖细胞有一定的保护作用。目前,有关HGF克服AML细胞动力学耐药临床研究结果表明,GM-CSF有可能提高患者的无病生存时间,副作用为血小板恢复延迟。显然,为明确HGF在克服AML细胞动力学耐药中的实际效果和可能的副作用,进一步进行HGF单独和联合条件下的大样本随机对照的前瞻性研究是十分必要的;另外,寻找能更有效的克服AML细胞周期耐药的HGF,也是应进一步研究的课题。     

骨髓增生异常综合征造血细胞生物学指标检测的意义

目的 研究骨髓增生异常综合征（MDS）造血细胞的生物学特性改变及与疾病进展的关系。方法对37例MDS患者骨髓细胞进行染色体核型分析、CFU－GM体外半固体琼脂培养、增殖细胞核抗原（PCR）表达、细胞周期分析以及造血克隆性检测,观察其对MDS疾病进展的关系。 结果 （1）MDS染色体异常检出率31％,随访过程中染色体核型正常的9例中有2例,异常的5例中有3例转化为急性髓系白血病（AML）。（2）MDS患者骨髓细胞体外CFU－GM集落产率减少、集簇产率增加。43％的患者CFU－GM产率极低或无生长。随访中9例CFU－GM集簇生长为主的患者中6例疾病进展或转化为AML;6例集落、集簇不生长和3例生长基本正常者中仅1例转化为急性白血病。（3）MDS患者骨髓细胞PCNA表达明显升高,RAEB／RAEB－t的PCN表达较RA／RAS高。（4）MDS患者骨髓细胞溴脱氧尿嘧啶（BrdU)标记指数减低（5．07％）,DNA合成期时间和潜在倍增时间延长,分别为8．57h和8．69d,并与病程进展关。（5）7例女性HUMARA等位基因为杂合子的RA患者中,2例为多克隆造血,5例为单克隆造血。3例RAEB和1例MDS／AML均为单克隆造血。结论 随着MDS病程进展,骨髓中原始细胞增多,造血细胞生物学特性呈现以下的演变趋势,造血转变为单克隆性;体外培养CFU－GM接近于白血病性生长特征;PCNA表达增高;细胞周期延长。     

Studies of Cellular Proliferation in Human Leukemia. I. Estimation of Growth Rates of Leukemic and Normal Hematopoietic Cells in Two Adults with Acute Leukemia Given Single Injections of Tritiated Thymidine

Two adults with rapidly progressive acute myeloblastic and myelomonoblastic leukemia were given single injections of tritiated thymidine, and measurements were made of the growth rates of their leukemic and normal hematopoietic cells by radioautographic methods. Although almost all leukemic blasts in both marrow and blood were metabolically active as shown by their ability to incorporate tritiated uridine and leucine in vitro, only 5.6% and 6.1% of the blasts in the marrow and even fewer in the blood incorporated tritiated thymidine. The mitotic indexes of the marrow blasts were 0.66% and 0.52%; no circulating blasts were dividing. The mean generation times of the actively proliferating blasts were estimated to be 49 and 83 hours. This cannot be equated with the doubling time of the total leukemic population as there is evidence that many blasts fail to continue dividing and die. The mean durations of the phases of the blasts' mitotic cycles were as follows: DNA synthesis (S) = 22 and 19 hours, premitosis (G(2)) = 3 hours, mitosis (M) = 0.47 and 0.62 hour (minimal estimates), and postmitosis (G(1)) = 24 and 61 hours. In both patients the maximal mean transit time of the blasts in the blood was 36 hours, and the minimal numbers of actively dividing blasts present were 1.6 and 2.6 x 10(9) per kg of body weight.Estimates were also made of the rates of proliferation and maturation of the residual normal erythrocytic and granulocytic cells in these two patients. Although total production was markedly diminished because of reduction in the number of normal elements, the relatively few remaining normal cells appeared to be dividing and maturing at rates that are about the same or only slightly slower than those found in normal subjects.We conclude that main reason leukemic blasts displace normal hematopoietic precursors in acute leukemia is that the blasts largely fail to differentiate. Many die but many others persist in the marrow and elsewhere as primitive cells and continue to proliferate. As the blasts accumulate, they gradually displace the normal hematopoietic cells, most of which continue their normal course of differentiation and leave the marrow as nondividing mature cells. It is not known why the over-all production of normal cells is not adequately increased to compensate for the anemia, granulocytopenia, and thrombocytopenia that develop, but apparently the leukemic cells somehow interfere with the proliferation or differentiation or both of normal stem cells.     

Prognostic potential of morphological and cytogenetic indices in patients with myelodysplastic syndrome

AIM: To examine prognostic potential of the number of bone marrow (BM) blasts and cell karyotype as risk factors of transformation of myelodysplastic syndrome (MDS) in acute myeloblastic leukemia AML. MATERIAL AND METHOD: The analysis of examination was made for 72 patients with primary MDS in the groups formed by number of blasts in BM, karyotype and IPSS variant. MDS was diagnosed by WHO criteria. Transformation into AML was established in blastosis > 20% in peripheral blood and/or BM. The karyotype was studied according to GTG technique. RESULTS: More frequent progression of MDS was seen in patients with blastosis > 10%, unfavourable karyotype and high IPSS risk. The least number of leukemic transformations occurred in karyotype of intermediate prognosis while disease-free survival in patients with karyotype of good prognosis was similar to that of patients with unfavourable karyotype. The number of blasts in BM and IPSS variant appeared to be prognostic markers of duration of leukemia-free survival in one-factor analysis. The multifactorial analysis found out one factor of MDS transformation in AML: number of blasts in BM puncture biopsy. CONCLUSION: Prognostic priority of the number of BM blasts as a risk factor of MDS progression compared to karyotype is explained by biological heterogenicity of MDS.     

Quantitative expression of Toll-like receptor-2, -4, and -9 in dendritic cells generated from blasts of patients with acute myeloid leukemia

BACKGROUND: Dendritic cells (DCs) generated from leukemic blasts constitute a promising tool in immunotherapy for acute myeloid leukemia patients (AML-DCs), because AML-DCs express human leukocyte antigens and costimulatory molecules such as CD40, CD80, and CD86 at a higher level than leukemic blasts. Potentiation of AML-DC vaccine might become feasible by the addition of adjuvants such as lipopolysaccharides (LPS) or CPG-rich oligodeoxyribonucleotides binding to Toll-like receptors (TLR) and inducing a stronger Type 1 T-cell response. STUDY DESIGN AND METHODS: mRNA and protein expression of TLR-2, -4, and -9 were analyzed with quantitative real-time polymerase chain reaction, Western blot, and flow cytometry for mature monocyte-derived DCs generated from 14 AML patients versus 14 healthy volunteers (HV-DCs), and the response of the AML- and HV-DCs to different microbial TLR ligands was determined by enzyme-linked immunosorbent assay for the proinflammatory cytokines tumor necrosis factor (TNF)-alpha, inducible protein (Ip)-10, and interleukin (IL)-6. RESULTS: AML-DCs and HV-DCs strongly expressed TLR-2 and TLR-4, while TLR-9 was expressed at a lower level in both groups. There was no significant difference in TLR expression between the two groups of AML-DCs and HV-DCs. In accordance with the TLR expression levels, DCs generated from both AML patients and HVs responded to the known microbial ligands peptidoglycan (PGN) and lipoteichoic acid for TLR-2 and LPS as ligand for TLR-4, by producing TNF-alpha and IL-6. A response to the ODNs 2006 and 2216 binding to TLR-9 was only detected in AML-DCs. CONCLUSION: Microbial ligands like ODNs and LPS constitute promising adjuvants for enhancing (AML-) DC vaccines.