THE LYMPHOCYTES OF CHRONIC LYMPHOCYTIC LEUKEMIA: THEIR PROLIFERATION AND CELL CYCLE KINETICS

Lymphocytes obtained from CLL patients exhibited a delayed and reduced response to PHA when cultured in diffusion chambers. DNA synthesis (8–10 hr) and general time (15–19 hr) of the late-developing CLL blasts were consistent with normal values ( T _s: 8–10 hr; T _c: 14–17 hr). However, the G₂ period of CLL blasts seemed more variable, and their mitotic index during the response at 5–6 days was 30–50% of the values determined for normal blasts during their peak response at 2–3 days.     

慢粒白血病中c-abl 癌基因的重排

作者采用α-³²P-dCTP标记的v-abl癌基因探针与7例慢粒白血病细胞DNA的PvuⅡ、Hind Ⅲ和Bg 1 Ⅱ酶切片段杂交。结果发现其中3例慢粒病例的Pvu Ⅱ和Hind Ⅲ酶切杂交图谱与正常对照相比有明显变化,且Pvu Ⅱ酶切位点有多处改变。表明在慢粒白血病中,Ph染色体重排亦可累及 c-abl癌基因内部,并可能有多次多点重排发生。     

KINETICS OF ERYTHROPOIETIN STIMULATED PROLIFERATION OF ERYTHROID PROGENITORS IN THE SPLEEN

The effect of RBC transfusion and erythropoietin (EPO) on the proliferation of immature erythrocyte progenitors was studied in the spleens of RBC transfused, lethally irradiated mice injected with bone marrow. Transfusion decreased expansion of the progenitors and slowed their proliferation: the mean cycle time as measured by per cent labelled mitosis (PLM) on the third day after injection of bone marrow was 10.7 hr in transfused as compared to 5.6 hr in non-transfused mice. One injection of five units of erythropoietin on day 2 decreased the mean cycle time to 7.3 hr in transfused mice and increased expansion of the progenitor cells. The effects of erythropoietin on cell proliferation were prompt: a significant increase of incorporation of ³H-TdR into DNA occurred within 2 hr of injection. Erythroblasts were absent from the spleens of transfused, irradiated bone marrow injected mice; however, erythroblasts appeared by 72 hr and 48 hr following EPO injection either 2 days or 5 days after transplantation respectively. Increased uptake of radioactive iron in spleen after erythropoietin injection preceded the appearance of erythroblasts by 2 and 1 days when erythropoietin was injected either 2 or 5 days after marrow transplantation respectively. The increase in cellular proliferation induced by erythropoietin in transfused irradiated mice injected with bone marrow equivalent to 0.35 femoral shaft was manifested as an increase of the total DNA content in the spleen by 119 μg (11.9 × 10⁶ cells) within 48 hr of injection. The cellular increment produced by EPO injection on day 5 to mice given 0.05 femoral shaft consisted mainly of undifferentiated mononuclear cells, most of which were labelled, with erythroblasts comprising only one quarter of the increment. Erythropoietin inactivated by mild acid hydrolysis failed to increase cellular proliferation.     

FINE STRUCTURAL ALTERATIONS OF INTERPHASE NUCLEI OF LYMPHOCYTES STIMULATED TO GROWTH ACTIVITY IN VITRO

This report describes fine structural changes of interphase nuclei of human peripheral blood lymphocytes stimulated to growth by short-term culture with phytohemagglutinin. Chromatin is found highly labile, its changes accompanying the sequential increases of RNA and DNA synthesis which are known to occur in lymphocyte cultures. In "resting" lymphocytes, abundant condensed chromatin appears as a network of large and small aggregates. Early in the response to phytohemagglutinin, small aggregates disappear during increase of diffuse chromatin regions. Small aggregates soon reappear, probably resulting from disaggregation of large masses of condensed chromatin. Loosened and highly dispersed forms then appear prior to the formation of prophase chromosomes. The loosened state is found by radioautography to be most active in DNA synthesis. Small nucleoli of resting lymphocytes have concentric agranular, fibrillar, and granular zones with small amounts of intranucleolar chromatin. Enlarging interphase nucleoli change chiefly (1) by increase in amount of intranucleolar chromatin and alteration of its state of aggregation and (2) by increase in granular components in close association with fibrillar components.     

ZINC REQUIREMENT FOR DNA REPLICATION IN STIMULATED HUMAN LYMPHOCYTES

The requirement for Zn⁺⁺ in DNA replication by phytohemagglutinin-stimulated human lymphocytes was studied. When 6 µM o-phenanthroline, a chelator with a high affinity for Zn⁺⁺, is added to cultures of stimulated lymphocytes a nearly complete inhibition of thymidine incorporation results within a few hours. In contrast, the incorporation of uridine is only slightly reduced and the incorporation of leucine is unaffected. m-Phenanthroline, a nonchelating analogue, does not alter the rate of thymidine incorporation even when present in 10-fold greater amounts than o-phenanthroline. The inhibition of thymidine incorporation by o-phenanthroline could be entirely reversed by the addition of Zn⁺⁺ to the cultures, or could be prevented by the prior addition of either Zn⁺⁺ or Ni⁺⁺. All other divalent cations tested were incapable of reversing the o-phenanthroline inhibition of thymidine incorporation.     

ABILITY OF THYMIC LYMPHOCYTES TO ALTER CFU KINETICS IN RADIATION CHIMERAS

It is known that the poor colony-forming ability of B6 bone marrow transplanted into B6D2F₁ hybrids can be improved if B6 lymphocytes are given in addition. It was recently reported that the augmenting lymphocytes decrease the doubling time of differentiating hemopoietic cells. to determine whether thymus cells alter the self-renewal of CFUs in this parent F₁ combination, retransplan-tation and ³H-thymidine ‘suicide’ were employed as methods to determine the cell-division rate. We have observed that in the presence of thymocytes, parental bone marrow cells are seeded more efficiently in the spleen, and the lag phase of the CFUs growth curve is shortened. However, thymic lymphocytes do not increase the slope of the exponential growth phase of CFUs.     

EXCLUSION OF SERUM FACTORS IN THE IN-VITRO DEPRESSION OF LYMPHOCYTES' RESPONSE TO MITOGENS IN CHRONIC LYMPHOCYTIC LEUKAEMIA

To find out whether serum factors are involved in the depression of the mitogenic response of lymphocytes of patients with chronic lymphocytic leukaemia, sera from these patients were used in the culture medium for normal lymphocytes. the results were considered on the basis of the dynamics of responses of these normal lymphocytes, comparing a standard human group AB serum with twenty sera from patients with CLL. No heat-stable inhibitory factors peculiar to CLL were found. In an experiment in which fresh CLL and normal sera were compared, no heat-labile inhibitory factors were apparent.     

L615白血病鼠胸腺和脾脏淋巴细胞脱氢酶同工酶的研究

应用聚丙烯酰胺凝胶电泳法对比研究了纯系６１５鼠和Ｌ６１５可移植性淋巴细胞型白血病鼠胸腺和脾脏淋巴细胞的葡萄糖－６－磷酸脱氢酶同工酶（ＥＣｌ．１．１．４９,Ｄ－葡萄糖－６－磷酸：ＮＡＤＰ氧化还原酶,Ｇ６ＰＤ）和乳酸脱氢酶同工酶（ＥＣｌ．１．１．２７,Ｌ－乳酸：ＮＡＤ氧化还原酶,ＬＤＨ）。并应用定量细胞化学法对ＬＤＨ和６ＰＤ全酶活性进行了测定。结果：在Ｌ６１５白血病鼠胸腺淋巴细胞中（白血病细胞占４０％）,Ｇ６ＰＤ同工酶谱显示异常,全酶活性明显增高、ＬＤＨ同工酶谱及全酶活性未发生明显变化。Ｌ６１５白血病鼠脾脏淋巴细胞中（白血病细胞占８４％）,Ｇ６ＰＤ和ＬＤＨ同工酶谱均显示异常,同时全酶活性也明显增强。提示：Ｇ６ＰＤ对白血病细胞的恶性增殖和浸润似乎更为敏感。     

Con A刺激致T淋巴细胞胞浆游离Ca~(2+)浓度升高

本文分别应用荧光Ca~(2+)指示剂Quin2和Indo-1研究了Con A刺激的T淋巴细胞[Ca~(2+)]i升高过程及其发生机制.结果表明Con A与T淋巴细胞作用可导致细胞[Ca~(2+)]i的迅速升高.这种增加的胞内游离Ca~(2+)不仅来自胞外Ca~(2+)的内流,也来源于胞内钙库的释放.其中Ca~(2+)内流与T细胞钙通道的开放有关.可被钙通道抑制剂戊脉胺抑制,细胞的去极化及钾通道阻断剂四乙胺均不能阻断Ca~(2+)的内流,提示Ca~(2+)内流不是通过电位操纵的钙通道实现的,也与拥通道的开闭无关.Ca~(2+)内流可能是通过Con A受体活化的受体操纵的钙通道而实现的.     

GRANULOPOIESIS AND CELL KINETICS IN CHRONIC MYELOID LEUKAEMIA

The present study is an analysis of the kinetic data for chronic myeloid leukaemia derived from personal observations and cases reported in the literature. A comparison is made with the parameters for normal granulopoiesis and the growth pattern of the CML cell population discussed.     

清栓酶在治疗慢性粒细胞白血病中的地位：附5例报告

慢性粒细胞白血病慢性期全血粘度,血浆粘度增高伴高粘滞综合症表现.在用马利兰作化疗同时应用清栓酶治疗,结果发现在马利兰未使白细胞下降前,患者全血粘度,血浆粘度明显下降及高粘滞综合症得以缓解,而其它血液流变学指标治疗前后也有明显改变.证明清栓酶能降低慢性粒细胞白血病化疗前血粘度增高,缓解高粘滞综合征临床表现.     

THE KINETICS OF CELL PROLIFERATION IN THE TRACHEOBRONCHIAL EPITHELIA OF RATS WITH AND WITHOUT CHRONIC RESPIRATORY DISEASE

Labelling indices of the tracheobronchial epithelia of conventionally-derived rats with chronic respiratory disease (CRD) and minimal-disease rats without CRD have been determined. The duration of the DNA synthesis phase (t_s) computed from the percentage of mitoses labelled at various intervals of time after injection of tritiated thymidine was 7 hr: t_G2 was 3.5 hr. Using the measured value of t_s and the labelling indices, the mean turnover times of the tracheobronchial epithelia in three groups of six 5-week-old conventionally-derived rats were calculated to be 11.2, 14.6 and 22.4 days, while in similar groups of 5-week-old minimal-disease rats the turnover times were found to be 24.3, 36.5 and 41.6 days. The majority of cell divisions in the tracheobronchial epithelium of these minimal disease rats were probably required for growth rather than renewal. The mean turnover time of this tissue in 5-week-old Syrian hamsters was 73 days. The cells of the rat tracheobronchial epithelium have been classed as basal or superficial, depending on their shape and proximity to the basement membrane. The mean turnover time of the basal cells in 5-week-old minimal-disease rats was 11.7 days calculated from labelling indices. The migration method of Brown & Oliver (1968) gave a similar value for the basal cells in minimal-disease rats, and a value of 9.5 days for the basal cells in a group of conventionally-derived rats. The mean turnover time in the latter was only 5.4 days if two rats with tracheobronchitis were included. Consideration of the slow rate of fall in mean grain count over labelled cells at intervals of time after labelling and the calculated turnover times suggests that the proliferative fraction of the basal cell population is close to unity. Well-labelled cells were still present in both basal and superficial populations in the minimal-disease rats at 10 days after labelling. The marked effects of CRD on cell proliferation in this epithelium are emphasized and the significance of this in relation to published work is discussed.     

白血病患者的肤纹研究

本文分析了l70名经骨髓检查确诊的白血病患者肤纹形态的手纹部分,数据经电子计算机进行判别分析,指纹类型的判对率为52.58±2.67％。根据掌褶类型判别时,其判对率为57±3.8％。指纹类型在判别急非淋患者及正常人时有显著性。掌褶类型的判别力对急非淋及慢粒患者有显著性。慢粒患者猿线出现率高于正常组。急淋患者右手悉尼线出现率高于正常组。此悉尼线在判别分析中有显著意义。急性白血病患者的弓斗指数高于对照组。白血病患者组掌纹白线的出现率也高于正常组。     

蟾蜍血液淋巴细胞姐妹染色单体互换及细胞周期动力学研究

本文对中华大蟾蜍Bufo bufo gargarizans血液淋巴细胞在植物血球凝集素(PHA)刺激条件下的细胞周期动力学及丝裂霉素C(MMC)诱发姐妹染色单体互换(SCE)的敏感性进行了研究。用姐妹染色单体差别染色方法确定细胞的分裂次数。实验结果表明,中华大蟾蜍血液淋巴细胞在体外培养2天后出现第二次分裂细胞,3天后达峰值(占分裂细胞中期相的38.9％),此后,其百分比逐日下降,培养后4天出现第三次分裂细胞,第5天出现第四次分裂细胞。Brdu浓度在8—24μg/ml范围内对SCE本底无影响。6ng/ml MMC所诱发的SCEs比对照组高3倍以上,说明中华大蟾蜍对MMC的诱变作用相当敏感。     

THE EFFECTS OF THROMBIN ON PHYTOHEMAGGLUTININ RECEPTOR SITES IN HUMAN PLATELETS

We have previously demonstrated that lentil phytohemagglutinin (lentil-PHA) binds to human platelet membranes without causing either aggregation or the release reaction. When platelets are treated with thrombin, there is an increase in lentil-PHA binding suggesting the appearance of new receptor sites on the cell surface. We prepared a lentil-PHA-ferritin conjugate using affinity chromatography which was used to saturate cell surface receptor sites. Studies using this conjugate suggest that thrombin causes a complex change in the platelet surface involving a decrease in the number of lentil-PHA receptor sites on the external platelet surface with a marked increase in sites within the center of the canalicular system. These increased sites may result from fusion of granule membranes with the canalicular membranes during the secretion process. There is no obvious relationship between lentil-PHA receptor sites and intramembranous particles.     

热休克T细胞中mRNA的表达

 富集的人外周血T淋巴细胞经40—42℃保温(热休克)可诱导产生90kd和71kd的两种主要热休克蛋白(HSP),此外,62和34kd HSP也可在不同条件下诱导。从不同处理的T淋巴细胞中提取mRNA并在免网织细胞裂解液系统中进行体外转译,显示出相同的主要HSP。其中71和62kdHSP不仅在升温时还可在低温(4℃)下诱导,表明淋巴细胞中几种主要HSP的诱导机制不完全相同。比较L-~(35)S-Met参入实验和体外转译的结果提示淋巴细胞中HSP基因表达主要在转录水平调控。     

靛玉红治疗慢性粒细胞白血病作用原理的研究

 靛玉红是一种新型抗癌药物,治疗慢性粒细胞白血病(CML)有较好的疗效。本文通过体内及体外实验采用靛玉红—脂质体为剂型,观察药物对CML细胞的作用。据荧光偏振度的测定结果表明,此药物分子能降低大白鼠白细胞膜流动性。还测定服药3天的患者外周白细胞中DNA聚合酶Ⅰ的活性,比治疗前降低74±1％。为靛玉红抑制CML细胞DNA合成的机理提供依据。此外,还使用载有靛玉红的脂质体进行体外实验,表明靛玉红可以直接降低细胞膜的流动性以及抑制CML细胞中DNA聚合酶Ⅰ活性。对靛玉红治疗CML的作用机理提出一些看法和讨论。