Growth of Human Lymphoma Cells in SCID Mice

Two EBV-negative human lymphoma cell lines raised in this laboratory and peripheral blood cells of a patient with large cell lymphoma in leukemic phase were injected intravenously or intraperitoneally into C.B.17 SCID mice. One line (OCI-LY18) was derived from the pleural fluid of a patient with a large cell, immunoblastic malignant lymphoma. Cells of this line are of B cell origin and characterized by multiple rearrangements of the JH locus. The second line (OCI-LY17) was grown from the peripheral blood of a patient with a large cell lymphoma of T-cell phenotype which has characteristic rearrangement of the T-cell receptor beta chain. The cells directly obtained from a patient with large cell non-cleaved malignant lymphoma were of B-cell origin.

Animals carrying OCI-LY18 developed large tumor masses within 6-8 weeks of inoculation. The tumors were detected in the intestine, mesentery, retroperitoneum, lymph nodes, spleen, lung and kidney. The masses resembled the primary tumor with respect to histological appearance and immunological phenotype. It was possible to generate secondary cell lines from the tumors found in the inoculated SCID mice. Injection of one of these secondary cell lines into SCID mice resulted in the rapid development of lymphoma and as few as 10 cells were sufficient to establish the disease in the inoculated animals. In contrast cells of OCI-LY17 produced small tumor aggregates that did not appear to progress over time and did not cause death of the animals. The tumors were identified by the same phenotypic profile as that seen in the primary cell line and sections of the patient derived lymph node biopsy.

The sample of peripheral blood cells directly derived from the patient gave rise to tumor nodules in multiple locations. Their morphology and immunological phenotype was consistent with the original disease seen in the patient.     

癌症患者群体LAK细胞的体外抗肿瘤的细胞毒活性

本研究应用~(125)I-UdR释放实验,测定了大量随机取样于111位癌症患者的外周血淋巴细胞以及从它们中诱导出的处于不同培养期的淋巴因子激活的杀伤细胞对K562和Raji肿瘤细胞的体外细胞毒活性.对560个有效数据的统计分析,获得以下结果:1.经过培养系统的作用,对K562细胞的细胞毒活性,从培养前的34.78±25%上升到8～13天培养期的68.04±17.3%之后基本稳定于近70%的水平,直至23～27天的培养期.此细胞毒活性的构成比模式表明,在培养之前,样品分布于很广的活性区域(10～90%);培养 8～13天和以后的培养期,约85%的样品集中到高活性区域(50～95%).2.对 Raji细胞的细胞毒活性,从培养前的8.9±8%上升到8～13天培养期的42.1±22%之后,在随后的培养期中,维持于约35%的水平.对Raji的细胞毒活性的构成比显示,在培养前,全部样品处于低活性区域(<25%);在培养过程中,部分样品(约30%)向高活性区域发生明显的扩展(50～90%),但另一部份样品(约40%)一直位于低活性区域(<35%).这样形成高低两个分布区域.这种现象的机理值得进一步探讨.     

去甲氧柔红霉素联合阿糖胞苷治疗急性髓系白血病的临床疗效

目的：探讨急性髓系白血病患者采用去甲氧柔红霉素联合阿糖胞苷治疗的临床疗效。方法70例急性髓系白血病患者按照随机数字表法分为对照组与治疗组,各35例。对照组采用柔红霉素治疗。治疗组采用去甲氧柔红霉素治疗。比较2组临床疗效、生存质量、毒副作用。结果治疗后治疗组治疗有效率为45．71％（16／35）与对照组[40．00％（14／35）]比较P＞0．05;治疗组治疗获益率为57．14％（20／35）,明显高于对照组[45．71％（16／35）]（P＜0．05）;2组患者KPS改善率比较（P＜0．05）。治疗组治疗期间不良反应发生率为48．57％（17／35）明显低于对照组[77．14％（27／35）]（P＜0．05）。结论去甲氧柔红霉素联合阿糖胞苷治疗急性髓系白血病患者的临床总体疗效较好,且显著改善患者生存质量,减少不良反应,值得推广应用。     

The Response of Peripheral Blood Stem Cells to Standard Chemotherapy for Lymphoma

Peripheral blood stem cells (PBSC) represent an alternative to bone marrow for autologous transplantation. These progenitors are found in the blood in greatly increased numbers after chemotherapy for acute leukaemia and after high dose cyclophosphamide when they can be harvested. We have assessed the response of colony forming units—granulocyte monocyte (CFU-GM) and colony forming units-granulocyte, erythroid, monocyte, megakaryocyte (CFU-GEMM) in 46 patients with lymphoma undergoing standard chemotherapy. Seventeen out of 18 patients with non Hodgkin's lymphoma (NHL) receiving CHOP had a 6.9 fold increase over baseline in CFU-GM and a 5 fold increase in CFU-GEMM occurring on day 19. In 3 patients with NHL, ALL type therapy induced a 17.5 and 16 fold increase in these progenitors occurring on day 21. Only 4 of the 10 patients with Hodgkin's disease (HD) treated with ChIVPP demonstrated a rise in progenitors and only to 3.3 and 2.7 fold in CFU-GM and CFU-GEMM respectively. ABVD similarly produced increases in only 5/9 patients and MOPP in 3/5. HOPE-bleo increased circulating CFU-GM 16.3 fold and CFU-GEMM 12.4 fold. Bone marrow involvement significantly reduced the peak levels of CFU-GM (p = 0.0017) as did second or subsequent line chemotherapy (p = 0.0001). Thus standard chemotherapy regimes for lymphoma can induce a rise in circulating progenitors in the majority of patients. Peripheral blood stem cell harvesting should be performed during first line chemotherapy as blood counts are recovering and should be repeated after each cycle.     

Light and Electron Microscopic Study of Vacuolated Cells in Immunoblastic Lymphadenopathy-like T-Cell Lymphoma

Three cases of immunoblastic lymphadenopathy (IBL)-like T-cell lymphoma were analyzed immunologically, and the ultrastructure of mononuclear cells in the lymph nodes and peripheral blood was examined. In the peripheral blood, light microscopic examination revealed vacuolated lymphocytes. These vacuolated lymphocytes formed rosettes with sheep erythrocytes, and they were CD3- and CD4-positive using the avidin-biotin method in cases 1 and 2. Electron microscopic examination revealed two kinds of abnormal lymphocytes. One kind was of B-cell nature with rich lamellated rough-surfaced endoplasmic reticulum and mitochondria. The other kind had a large cytoplasm, in which Golgi apparatus, some endoplasmic reticulum, some mitochondria and a few vacuoles were seen. Some of these vacuoles had remnants of mitochondrial cristae or were enlarged endoplasmic reticulum. The vacuolated T lymphocytes and activated lymphocytes of B-cell nature disappeared with chemotherapy but reappeared with relapse of the disease. These observations suggest that vacuolated cells are related to pale cells in lymph node sections. In other words when these vacuolated cells are found in the peripheral blood of patients with lymphoid malignancies, IBL-like T-cell lymphoma can be suspected.     

Transport Mechanisms of Idarubicin, an Anthracycline Derivative, in Human Leukemia HL60 Cells and Mononuclear Cells, and Comparison with Those of Its Analogs

Transport mechanisms of idaruhicin (IDA) in HL60 cells, as leukemia cells, and human mononuclear cells (MNCs), as normal cells, were investigated, and compared with those of its analogs. The uptake of IDA by both cell types was temperature- and concentration-dependent, was inhibited competitively by daunorubicin (DNR) and noncompetitively by adriamycin (ADR), and was stimulated by preloading of the cells with DNR and ADR, indicating the partial involvement of a carrier-mediated mechanism. On pretreatment of the cells with 2,4-dinitrophenol, IDA uptake by HL60 cells increased, but that by MNCs decreased, suggesting that IDA was partially taken up into HL60 cells via an energy-independent carrier system, and into MNCs via an energy-dependent one. We speculated that in HL60 cells the carrier concerned with IDA uptake was common to DNR and ADR, and that the binding site of IDA on the carrier was the same as that for DNR, but not that for ADR, while in MNCs the carrier system consisted of, at least in part, a carrier for DNR uptake and one for ADR uptake, and the binding site of IDA was identical to that for DNR in the former, but different from that for ADR in the latter. It appeared that the uptake of IDA was greater than those of pirarubicin, DNR and ADR in both HL60 cells and MNCs, and that IDA was incorporated into MNCs more efficiently than into HL60 cells because of the higher uptake efficacy of the carrier(s).     

Idarubicin and High Dose Cytarabine: A New Salvage Treatment for Refractory or Relapsing Non-Hodgkin's Lymphoma

Twenty three patients with relapsing (n = 11) or refractory (n = 12) non-Hodgkin's lymphoma (NHL) to one or two prior anthracycline based combination chemotherapy regimens were treated as second or third line regimen with 3 induction cycles of Idarubicin (IDA) (7 mg/m²/d IV dl-d3) and high dose cytarabine (HD Ara-C) (1 g/m²/12 h IV dl-d3), each cycle was repeated every 3 weeks. Responding patients received a maintenance therapy with monthly cycles of IDA : 15 mg/m² dl-d3, Etoposide 100 mg/m² dl-d3, both by oral route. Twenty two patients are evaluable and we observed 13 CR and 1 PR with an overall response rate of 61 % (14/23; 95% Cl = 38.5% 80.3%). The median time to progression was 32 months (6.5 - 63 + m.). The response rate to IDA-HD Ara C was not different for patients with (n = 14) or without (n = 9) objective response to the last prior therapy. The main toxicity was hematological: all patients experienced grade 4 neutropenia and 22 patients had grade 4 thrombopenia, but there were no toxic deaths. IDA and HD-Ara-C combination is highly effective in refractory or relapsed NHL. As hematological toxicity was the limiting factor for further escalation of dose-intensity, further studies might include hematopoietic growth factors support in the therapeutic scheme.     

薏苡仁提取物对原发性肝癌细胞毒性作用的实验研究

目的 :研究康莱特注射液对原发性肝癌细胞的毒性作用。方法 :用MTT法对康莱特等 7种药物的体外抗肝癌细胞活性进行检测。结果 :康莱特与常用的化疗药CP、DDP、5 FU等对肝癌细胞有毒性作用 ,可达 30 %以上。结论 :康莱特注射液对肝癌细胞有较好的毒性作用 ,是一种天然的副作用低的抗癌新药 ,有良好的应用前景。     

Apolloon反义寡核苷酸对K562细胞增殖和凋亡的影响

目的探讨Apollon反义寡核苷酸（antisense oligonucleotide,ASODN）对白血病细胞K562增殖及凋亡的影响。方法设计合成特异性的Apollon硫代磷酸ASODN及其对照错义寡核苷酸（missense oligonucleotide,MSODN）,脂质体介导转染K562细胞后,采用MTT法检测K562细胞增殖,Annexin V-FITC法检测细胞的凋亡。结果Apollon反义寡核苷酸对K562细胞的生长抑制呈浓度-时间依赖性。Apollon ASODN在终浓度为600 nmol/L作用K562细胞时,能明显抑制其增殖,K562细胞凋亡率显著增加（P<0.01）。结论Apollon 反义寡核苷酸可下调Apollon基因表达,有效抑制K562细胞的增殖,并促进K562细胞凋亡。     

Plasma Pharmacokinetics of Idarubicin and its 13-Hydroxymetabolite after Intravenous and Oral Administration under Fasting and Non-Fasting Conditions

The plasma pharmacokinetics of Idarubicin and its 13-hydroxymetabolite have been studied in 10 patients with solid tumours after intravenous and oral administration under fasting and non-fasting conditions in a randomized cross-over design. The plasma concentration time curves of Idarubicin after intravenous administration could be described by the open two or three compartment models. No pharmacokinetic modelling of Idarubicin was possible after oral administration. After oral administration of Idarubicin, the amount of intact drug was higher under non-fasting conditions. The extensive and long-lasting appearance of Idarubicinol suggests that this cytotoxic metabolite is of major clinical importance in i.v. and oral therapy with Idarubicin. The pharmacokinetics of Idarubicinol was not affected by food intake.     

嵌合寡核苷酸对端粒酶活性的抑制作用

目的： 研究嵌合寡核酸对端粒酶活性的抑制作用。 方法： 合成各种寡核苷酸（ODNs）,利用HL60细胞溶解液实施体外实验，U87细胞系观察这些寡核苷酸在细胞水平的作用。 结果： 硫代磷酸酯寡核甘酸（PSODNs）和RNA模板反义ODNs都可以抑制端粒酶的活性，且可产生累积效应。PSODNs的3′端延长一段与端粒酶RNA模板区域互补的各种修饰的ODNs则可以更进一步地提高它们的抑制效率。 结论： 研究设计的嵌合反义寡核苷酸（cODNs）作为目前较有效的端粒酶活性抑制剂，可成为肿瘤治疗中较有前途的药物之一。     

The Mechanisms of Cytotoxicity to Tumor Cells by Polymorphonuclear Leukocytes Stimulated with Cytokines

The mechanisms of tumor cytotoxicity of rat polymorphonuclear leukocytes (PMN) activated with cytokine(s) were studied with the use of supernatants from a rat myelomonocytic leukemia cell line, WRT-7, incubated in the presence of bacterial lipopolysaccharide (LPS) (LPS WRT-7 sup) as a source of cytokine. Rat peritoneal PMN treated with LPS WRT-7 sup showed cytostasis from 3 hr after the start of incubation, while significant cytolysis was first observed after 24 hr. When target tumor cells were separated from PMN at 6 or 12 hr after the start of the assay, 3H-UdR release from the separated target cells comparable to that from the group incubated with PMN for the whole assay time of 40 hr was observed during the following incubation, which indicates that priming for subsequent lysis occurs at a relatively early stage of the assay. None of various scavengers of active oxygens, inhibitors of heme enzymes, and inhibitors of neutral proteinases inhibited cytolysis mediated by PMN stimulated with LPS WRT-7 sup. Heparin inhibited PMN cytolysis only when it was added within 1 hr after the start of the assay. Fractionation of heparin by ion exchange chromatography showed a parallelism between the negative charge and the inhibitory effect of heparin on PMN cytotoxicity.     

Efficacy and Toxicity of Idarubicin Versus High-dose Daunorubicin for Induction Chemotherapy in Adult Acute Myeloid Leukemia: A Systematic Review and Meta-analysis

Background

The 2 main formulations of anthracycline used for acute myeloid leukemia (AML) induction therapy are idarubicin (IDA) and daunorubicin.

Bcl-2 Antisense Oligonucleotide Inhibits the Proliferation of Childhood Leukemia/lymphoma Cells of the B-cell Lineage

An 18-mer phosphorothioate bcl-2 atisense oligonucleotide (ASO) inhibited colony formation of three B-cell leukemia/lymphoma cell lines in a dose dependent manner in the range of 0.125–0.5 μmol/l. The srcambled cogener had no detectable effect. A decrease in BCL-2 protein and apoptotic DNA fragmentation was detected in the studied cell lines and primary blast cells of two children with acute lymphoblastic leukemia. Neither BCL-2 protein level, nor DNA integrity was affected by the scrambled control indicating the specific effect ASO. As far as we know, this is the first report on the effects of bcl-2 ASO on childhood leukemia/lymphoma cell samples.     

Idarubicin containing regimen in multiple myeloma: preliminary results of a pilot study using a modified "TANDEM" transplant program

Tandem autologous transplant actually represents a challenge in multiple myeloma treatment, but the best conditioning regimen is still under investigation. With the aim of evaluating the feasibility of a modified tandem transplant strategy, we treated 10 multiple myeloma patients after conventional first line chemotherapy with a two step conditioning regimen consisting of high-dose melphalan (200 mg/m 2 ) followed by high-dose melphalan (180 mg/m 2 ) together with indarubicin (15 mg/sqm 2 c.i. ×3 days) both with peripheral stem cell support. At first transplant, the median age was 62 years, performance status was good and disease status was CR in 2 patients and PR in the rest. At the end of the first transplant, 70% of patients achieved CR and only mild toxicity was observed. After the second transplant further improvement of the response rate was obtained with 90% CR. However, we observed three toxic early infection-related deaths from CMV and legionella pneumonia at day +17, +26, +54 after transplantation. Although this schedule seems to be effective in terms of response rate, the 30% TRM imposes an anthracycline dose-reduction with careful patient selection. This approach could reduce the toxic effects and maintain the efficacy of therapy at the same time.     

Agar Capillary Clonogenic Microassays for Cellular Immunocytotoxic Activities in Human Leukaemia and Lymphoma

Current concepts of immunotherapeutic approaches in Ieukemias and lymphomas using activated cytotoxic lymphocytes and macrophages are briefly reviewed. Defective cellular immunocytotoxic activities and effects of interleukins and chemotherapeutic drugs thereupon are discussed. In vitro assays to measure lymphokine-activated killer (LAK) and natural killer (NK) cell activities suffer from various problems, depending on the quality of the endpoints. Our clonogenic microassay for LAK cell activity, using agar-containing glass capillaries, avoids some of the potential artifacts and offers several advantages that are discussed. As an example the stimulatory effect of low mafosfamide concentrations on the LAK cell activity versus K562 human myeloid Leukemia cells is demonstrated. Thus, our clonogenic LAK microassay provides a valid tool for preclinical screening of immunomodulatory agents.     

Effect of Cytotoxic Cells on Minimal Residual Leukemia

The presence of minimal residual disease is indicated by the high frequency of relapses after twin bone marrow transplants and after allogeneic bone marrow transplants without graft versus host disease (up to 75% and 45% of cases, respectively). The graft versus leukemia effect may be mediated by IL-2 activation of natural killer cells (CD16 +, CD56 +, CD3-, CD8+/-) or cytotoxic T cells (CD3 +, CD56+/-). These activated killer cells can bind to targets and cause their lysis, and then recirculate to kill other targets. Killing can be blocked by anti-perforin antibodies and enhanced by protein kinase C-activation of effectors

There are several studies indicating that a high percentage of leukemic cells can be killed by LAK-cells. However even in the most sensitive cases, lysis of all the cells cannot be achieved. The finding that leukemic clonogenic cells are generally sensitive to both NK and LAK cytotoxicity provides a more hopeful possibility

The lack of tumor specific antigents, leukemic cell-immunoheterogeneity and maturation asynchrony explains why antigen dependent T-cell mediated cytotoxicity is only partly effective in eradicating residual leukemic cells. Future work should therefore include more studies of the mechanism of resistance to LAK cells, possibilities of further enchancing cytotoxicity and the mechanism of graft-versus-leukemia effect     

离体时间对LAK细胞前体及LAK活性的影响

作者测定人胚胎胸腺、脾脏ＬＡＫ前体和同种异体ＬＡＫ细胞前体在离体后不同时间内的活力及相应的ＬＡＫ毒活性。