The measurement of ferritin in the leukemic blasts with a "sandwich" type enzyme immunoassay method

The concentration of ferritin in serum and leukemic blasts were measured with the improved method of enzyme immunoassay for ferritin which uses beta-D-galactosidase from Escherichia coli. Precise measurement of ferritin was possible for concentration as low as 0.2 ng/ml serum. A good correlation (correlation coefficient, 0.946) existed between the ferritin radioimmunoassay and enzyme immunoassay. The serum ferritin levels in healthy males were 97.2 +/- 46.7 ng/ml, whereas those in healthy females were 53.5 +/- 27.1 ng/ml. Serum ferritin level was markedly increased (698 +/- 447 ng/ml) in sera of 36 patients with various types of acute leukemia, of which acute monocytic leukemia showed the highest serum ferritin concentrations (1156 +/- 324 ng/ml). Leukemic blasts from 24 patients with acute leukemia contained markedly elevated concentrations of ferritin (64.3 +/- 38.7 ng/10(6) cells) as compared with the ferritin concentration in normal leukocytes (7.1 +/- 2.8 ng/10(6) cells). The blasts cells from the patients with acute monocytic leukemia showed the highest ferritin levels (105 +/- 26.6 ng/10(6) cells).     

The effect of high-dose methylprednisolone treatment on GM-CSF level in children with acute leukemia: a pilot study.

High-dose methylprednisolone therapy (HDMP) induces acceleration of leukocyte recovery in acute lymphoblastic leukemia (ALL) and the differentiation of myeloblasts to mature granulocytes in acute myeloblastic leukemia (AML). These effects of corticosteroids have been shown to be due to the enhanced colony-stimulating activity (CSA) and responses to corticosteroids in some patients with aplastic anemia and myelodysplastic syndromes (MDS) have been related to increased CSA activity. We measured the serum (granulocyte-macrophage colony-stimulating factor (GM-CSF) levels by a sandwich linked immunoabsorbent assay (ELISA) in patients with ALL and AML at presentation and following high-dose methylprednisolone (HDMP) therapy. Serum GM-CSF levels at presentation in the ten cases studied ranged between 160 and 700 pg/ml (mean 418.5 +/- 252.5). One week following HDMP therapy GM-CSF levels increased to between 260 and 950 pg/ml (733.5 +/- 203.2). Four weeks after therapy the GM-CSF levels increased to between 470 and 1350 pg/ml (911 +/- 278.7). GM-CSF levels were markedly elevated one week after HDMP in the patients with ALL, suggesting that in addition to the lymphotoxic effects on leukemic blasts, the acceleration in neutrophil recovery may be due to release of GM-CSF induced by HDMP and its effects on myeloid progenitors.     

Determination of serum soluble macrophage colony-stimulating factor receptor levels in patients with hematological diseases

Many cytokine receptors are synthesized as secreted soluble forms. The soluble receptors are created either by proteolytic cleavage of membrane-bound receptor to release extracellular portion, or by alternative mRNA splicing. Soluble cytokines exert their effects through the action of competing with membrane-bound receptors for binding to their ligand, or the formation of ligand- receptor complex to protect the ligand from degradation and contribute to the enhanced half-life of the ligand. In…     

Potential use of serum CD44 as an indicator of tumour progression in acute leukemia

CD44 is a widely expressed cell surface glycoprotein with various functions. This molecule is shed from the cell surface and released as a soluble molecule. High serum levels of CD44 have been demonstrated in some solid tumours. In this study we measured serum CD44 in 25 patients with acute leukemia, in 12 with bacterial infections, and in 13 normal controls. The levels of serum CD44 of patients with bacterial infections were significantly higher (mean 531.3+/-60.1 ng/ml, p<0.001) than those of normal controls (299. 0+/-115.4 ng/ml). Acute leukemia patients before treatment had almost four-fold higher levels of serum CD44 than normal controls (mean 1301.9+/-1384.6 ng/ml, p<0.01). Serum CD44 levels were correlated with clinical status. After treatment the serum CD44 levels significantly decreased, but they were still higher than in normal controls. Patients in complete remission all relapsed if serum CD44 levels were higher than 500 ng/ml (normal+2 SD) after chemotherapy. The serum CD44 levels were correlated with the absolute numbers of leukemic cells in peripheral blood. The results demonstrated that serum CD44 levels correlate well with the clinical status of acute leukemia, and such evaluation may provide a reliable tumour marker of acute leukemia.     

急性白血病患者血清中期因子定量检测及其临床意义

目的 探讨血清中期因子(MK)水平在急性白血病(AL)不同发展阶段的意义及其与WT1基因的关系.方法 用酶联免疫吸附实验(ELISA)检测86例初治和完全缓解的AL患者组及30名健康对照血清中MK水平;用实时定量PCR法检测其中15例初治急性髓系白血病(AML)患者骨髓细胞中WT1表达量.结果 初治组血清MK水平(中位数及四分位数)为7.52 ng/ml[(5.44,10.55)ng/ml],缓解组MK水平为3.52 ng/ml[(1.56,5.20) ng/ml],健康对照组MK水平为2.44 ng/ml[(1.89,3.12)ng/ml],初治组较缓解组和健康对照组水平均升高,差异有统计学意义(P＜0.01),缓解组和健康对照组间MK水平差异无统计学意义(P＞0.05).初治B细胞急性淋巴细胞白血病(B-ALL)组血清MK水平为7.88 ng/ ml[(5.78,15.78) ng/ml],初治AML中MK水平为6.25 ng/ml[(4.59,16.33) ng/ml],两组间差异无统计学意义(P＞0.05);血清MK水平与骨髓中WT1基因表达量间存在相关性(r=0.529,P=0.043).结论 初治AL患者血清中MK表达水平升高,完全缓解后MK表达水平下降,ELISA法测定血清MK可以作为AL患者病情评价的手段之一;MK与WT1在基因定位上相邻,临床意义相近,并且血清MK和WT1基因在定量上存在相关性.     

超重及肥胖急性白血病患者血清中脂肪素表达的研究

目的 研究肥胖、超重对急性白血病患者血清中脂肪素的表达及预后的影响.方法 收集2008年7月至2015年7月在辽宁省人民医院和中国医科大学附属盛京医院治疗的急性白血病患者570例,根据体质量指数(BMI)分为肥胖/超重组(BMI≥24 kg/m2)和正常体质量组(BMI<24 kg/m2),采用双抗体夹心亲和素-生物素复合酶联免疫吸附试验(ABC-ELISA)检测脂肪素水平,t检验比较两组间血清脂联素、 瘦素和抵抗素表达水平的差异;Kaplan-Meier法比较两组患者间生存情况的差异.结果 急性淋巴细胞白血病(ALL)和急性髓系白血病(AML)肥胖/超重组血清脂联素表达水平均低于正常体质量组[ALL成年组(3.8±2.1)pg/ml比(6.4±2.9)pg/ml,儿童组(4.2±2.7)pg/ml比(7.4±3.1)pg/ml);AML(4.1±2.3)pg/ml比(6.9±3.1)pg/ml](t值分别为-2.291、-2.462、-2.244,均P<0.05);ALL儿童患者肥胖/超重组瘦素表达水平较正常体质量组升高[(34±17)pg/ml比(21±17)pg/ml,t=2.092,P=0.038];所有急性白血病患者肥胖/超重组与正常体质量组间抵抗素表达水平比较,差异均无统计学意义(均P>0.05).Kaplan-Meier生存分析结果显示,AML和ALL成年患者肥胖/超重组5年总生存率均低于正常体质量组(38.0％比46.6％,χ2=1.449,P=0.001;41.4％比48.4％,χ2=4.166,P=0.041).结论对于急性白血病患者,肥胖、超重者血清脂联素水平降低,肥胖、超重的成年ALL及AML患者的5年总生存率较正常体质量患者低.     

Superoxide radical generation and Mn- and Cu-Zn superoxide dismutases activities in human leukemic cells

Mn- and Cu-Zn superoxide dismutase (SOD) activities and generation of superoxide radicals (O(2) (-)) were assessed in leukemic cells from 10 patients with acute myeloid or monocytic leukemia (AML) and 10 patients with acute lymphoblastic leukemia (ALL), using a sensitive, specific chemiluminescence method. Leukemic cells were classified according to the French-American-British classification. M4 AML cells from two patients produced some O(2) (-) upon stimulation with opsonized zymosan (OZ), phorbol myristate acetate (PMA) or N-formyl-methionyl-leucyl-phenylalanine (FMLP), but less than normal granulocytes or monocytes. M5b AML cells from one patient produced as much O(2) (-) in response to these stimulants as normal monocytes. No O(2) (-) generation was induced in other types of leukemic cells. Total SOD activity in AML cells was significantly greater in normal granulocytes, but was only half of the activity in ALL cells. Mn-SOD in AML cells was very low or undetectable. These results suggest that except in M5b cells, decreased O(2) (-) production may contribute to susceptibility to infections in AML patients. Decreased Mn-SOD activity in AML cells may predispose them to oxidative stress.     

Clinical Significance of Serum p53 and Epidermal Growth Factor Receptor in Patients with Acute Leukemia

Background: Pretreatment serum p53 and epidermal growth factor receptor (EGFR) were assessedusing enzyme-linked immunosorbent assay (ELISA) in patients with acute leukemia to analysis their roles incharacterization of different subtypes of the disease. Materials and Methods: Serum samples from thirty twopatients with acute myeloid leukemia (AML) and fourteen patients with acute lymphoid leukemia (ALL) wereanalysed, along with 24 from healthy individuals used as a control group. Results: The results demonstrateda significant increase of serum p53 and EGFR in patients with AML (p<0.0001) compared to the controlgroup. Also, the results showed a significant increase of both markers in patients with ALL (p<0.05, p<0.0001respectively). Sensitivities and specificities for these variables were 52% and 100% for p53, and 73.9%, 95.8%for EGFR. Serum p53 and EGFR could successfully differentiate between M4 and other AML subtypes, whilethese variables failed to discriminate among ALL subtypes. A positive significant correlation was noted betweenp53 and EGFR. Negative significant correlations were observed between these variables and both of hemoglobin(Hg) content and RBC count. Conclusions: Mutant p53 and EGFR are helpful serological markers for diagnosisof patients with AML or ALL and can aid in characterization of disease. Moreover, these markers may reflectcarcinogenesis mechanisms.     

抗原处理相关转运因子在小儿急性白血病患者中的表达及其临床意义

背景与目的:抗原处理相关转运因子(transporterassociatedwithantigenprocessing,TAP)参与免疫监视,因而可能与肿瘤发生有关。本文旨在探讨急性白血病TAP分子表达及其临床意义,探讨急性白血病的治疗策略。方法:采用RT-PCR检测34例初治急性淋巴细胞白血病(acutelymphoblasticleukemia,ALL)(初治组)、15例复发ALL(复发组)及20例急性髓系白血病(acutemyeloidleukemia,AML)患者骨髓中TAP亚单位TAP1和TAP2的表达。20例无全身性疾病外科住院患儿作为对照组。使用数码成像分析仪测定并计算扩增条带的相对于阳性内对照GAPDH的吸光度(A)值。结果:ALL初治组和ALL复发组的TAP1A值(分别为0.448±0.167和0.169±0.021)及TAP2的A值(分别为0.196±0.180和0.112±0.020)均低于对照组,P均<0.01;AML组TAP2的A值低于对照组(P<0.01);ALL复发组TAP1的A值低于ALL初治组,P<0.05;ALL初治组复发者(6/34)TAP1的A值(0.215±0.159)较持续完全缓解(constantcompleteremission,CCR)者低(24/34,0.462±0.189,P<0.05)。结论:小儿ALL和AML均存在TAP分子低表达,这可能促使白血病细胞逃避免疫监视;TAP1亚单位低表达可能与ALL的复发有关。     

Circulating levels of thrombopoietic and inflammatory cytokines in patients with acute myeloblastic leukemia and myelodysplastic syndrome

BACKGROUND: The regulation of megakaryocytopoiesis and thrombopoiesis appears to be under the control of an array of hematopoietic growth factors. The regulatory mechanism of endogenous cytokines in circulating platelet counts of thrombocytopenic patients with acute myeloblastic leukemia (AML) and myelodysplastic syndrome (MDS) is still not clear. METHODS: We measured the serum levels of both thrombopoietic and inflammatory cytokines in peripheral blood and bone marrow samples collected from 52 patients with either AML or MDS along with 35 normal control samples. The levels of thrombopoietin (TPO), interleukin (IL)-11, IL-6, IL-8 and stem cell factor (SCF) were determined by ELISA. RESULTS: Platelet counts in the AML/MDS patients during initial diagnosis, chemotherapy and complete remission were 71.2 +/- 11.6, 47.2 +/- 6.1 and 181.4 +/- 26.3 x10(9)/l, respectively. The median value of TPO in AML/MDS patients during diagnosis was 150.6 pg/ml and increased significantly during chemotherapy (median: 828 pg/ml; p < 0.05) but then decreased following complete remission (median: 221.4 pg/ml). However, these levels were all significantly higher in patients than in normal subjects (p < 0.05, p < 0.05 and p < 0.05; respectively), and no significant change was noted in the levels of IL-11 and SCF during treatment of patients or in normal controls. The level of IL-6 was not detectable in normal serum samples but was markedly increased in the AML/MDS patients (median level during diagnosis: 6.7 pg/ml; chemotherapy: 25 pg/ml; complete remission: 7 pg/ml). The level of IL-8 in patients with AML and MDS was markedly elevated during diagnosis (median: 27.5 pg/ml; range: 0-1,587 pg/ml), but decreased to the level of the normal controls when patients were under chemotherapy or in complete remission. CONCLUSIONS: The endogenous levels of TPO, IL-6 and IL-8 are elevated in the thrombocytopenic patients with AML and MDS. Our results are consistent with previous mechanistic studies and suggest that TPO and IL-6 may be active mediators of platelet production.     

Plasma levels of leptin and soluble leptin receptor and polymorphisms of leptin gene -18G > A and leptin receptor genes K109R and Q223R,in survivors of childhood acute lymphoblastic leukemia

Background

Approximately 20% of children and adolescents in Europe are overweight. Survivors of pediatric acute lymphoblastic leukemia (ALL) are at increased risk of overweight and obesity. The purpose of this study was to assess leptin and leptin soluble receptor levels, as well as polymorphisms of selected genes in survivors of pediatric ALL, and the influence of chemo- and radiotherapy on development of overweight in the context of leptin regulation.

Methods

Eighty two patients (55% males), of median age 13.2 years (m: 4.8 years; M: 26.2 years) were included in the study. The ALL therapy was conducted according to modified Berlin-Frankfurt-Munster (BFM; n = 69) regimen or New York (n = 13) regimen. In 38% of patients cranial radiotherapy (CRT) was used in median dose of 18.2Gy (m: 14Gy; M: 24Gy). Median age at diagnosis was 4.5 (m: 1 year; M: 16.9 years) and median time from completion of ALL treatment was 3.2 years (m: 0.5 year; M: 4.3 years). Patients with BMI ≥85 percentile were classified as overweight. Correlation of plasma levels of leptin and leptin soluble receptor, and polymorphisms of leptin gene -18G > A, leptin receptor genes K109R and Q223R, and the overweight status were analyzed in relation to gender, intensity of chemotherapy (high intensity vs. standard intensity regimens) and to the use of CRT.

Results

Significant differences of leptin levels in patients treated with and without CRT, both in the entire study group (22.2+/- 3.13 ng/ml vs. 14.9+/-1.6 ng/ml; p < 0.03) and in female patients (29.9+/-4.86 ng/ml vs. 16.9+/-2.44 ng/ml; p = 0.014), were found. Significant increase of leptin levels was also found in overweight patients compared to the non-overweight patients in the entire study group (29.2+/-2.86 ng/ml vs. 12.6+/-1.51 ng/ml; p < 0.0001), female patients (35.4+/-6.48 ng/ml vs. 18.4+/-2.5 ng/ml; p = 0.005), and male patients (25.7+/-2.37 ng/ml vs. 6.9+/-0.95 ng/ml; p < 0.0001). Negative correlation was observed for plasma levels of soluble leptin receptor and overweight status, with significant differences in overweight and non-overweight patients, both in the entire study group (18.2+/-0.75 ng/ml vs. 20.98+/-0.67 ng/ml; p = 0.017) and in male patients (18.2+/-1.03 ng/ml vs. 21.8+/- 1.11 ng/ml; p = 0.038). Significant (p < 0.05) negative correlation was found between leptin and leptin receptor levels in the entire group (correlation coefficient: 0.393) and in both gender subgroups (correlation coefficient in female patients: -0.427; in male patients: -0.396).

Conclusions

The prevalence of overweight in our cohort was higher than in general European population (31% vs 20%) and increased regardless of the use of CRT. Leptin and leptin receptor levels may be used as useful markers of high risk of becoming overweight in ALL survivors, particularly in females treated with CRT. Polymorphisms of leptin gene -18G > A and leptin receptor genes K109R and Q223R were not associated with overweight status in ALL survivors.     

Levels of the soluble, 55-kilodalton isoform of tumor necrosis factor receptor in bone marrow are correlated with the clinical outcome of children with acute lymphoblastic leukemia in first recurrence

BACKGROUND: It has been shown that the soluble, 55-kilodalton isoform of tumor necrosis factor receptor (sTNFRp55) enhances tumor survival by exhibiting competitive ligand binding. The objective of the current study was to determine the levels of sTNFRp55 and their impact on outcome in 106 children with acute lymphoblastic leukemia (ALL) in first recurrence. METHODS: Between January 1997 and December 2001, bone marrow (BM) samples were collected from 106 children with a first recurrence of ALL at diagnosis. These patients were enrolled in the Berlin-Frankfurt-Münster (BFM) ALL recurrence trial, ALL-REZ BFM 90-96. Levels of sTNFRp55 in BM samples were determined with a commercially available enzyme-linked immunosorbent assay kit. Event-free survival (EFS) and overall survival were assessed from the date of study entry or the date of randomization, as appropriate. RESULTS: The mean sTNFRp55 level (+/- standard deviation) was 3.40 +/- 2.57 ng/mL. High levels of sTNFRp55 were associated with shorter duration of first complete remission and observation time as well as poor response to chemotherapy. Most importantly, the probability of EFS (pEFS) at 3 years was significantly worse for children with recurrent ALL who had sTNFRp55 levels greater than the median value (> 2.77 ng/mL) compared with patients who had levels that were less than the median value (pEFS: 0.44 +/- 0.10 ng/mL vs. 0.12 +/- 0.10 ng/mL; P = 0.006). It is noteworthy that the sTNFRp55 levels in 22 children with recurrent, TEL-AML1-positive ALL ([t(12;21)(p13;q22)]; 2.69 +/- 1.05 ng/mL) were significantly lower compared with the levels in children who had TEL-AML1-negative ALL (3.34 +/- 1.49 ng/mL; P < 0.05). CONCLUSIONS: The results indicated that a high sTNFRp55 level represents a negative prognostic factor for children with recurrent ALL in terms of EFS and overall survival.     

Hepatocyte growth factor levels in bone marrow plasma of patients with leukaemia and its gene expression in leukaemic blast cells.

Hepatocyte growth factor (HGF) has been known as a multiple function factor, which also stimulates early haematopoiesis. In this study, we found that HGF was expressed at both the RNA and protein levels in acute myeloid leukaemia (AML) and chronic myeloid leukaemia (CML). In patients with AML (n = 20) and CML (n = 5), bone marrow plasma HGF concentrations were 20.44 +/- 6.26 (mean +/- s.e.) ng ml-1 and 7.17 +/- 0.53 ng ml-1 respectively. These were significantly higher (P < 0.01) than the value for normal subjects (n = 26): mean 0.92 +/- 0.09 ng ml-1. Constitutive HGF production was observed in freshly prepared leukaemic blast cells from three patients with high HGF levels of bone marrow plasma. Expression of HGF mRNA was correlated with bone marrow plasma HGF levels. After complete remission was obtained in six patients, bone marrow plasma HGF levels were significantly decreased. In contrast, the HGF mRNA was less abundantly expressed in acute lymphoid leukaemia (ALL). In patients with ALL (n = 5), bone marrow plasma HGF concentration (0.69 +/- 0.14 ng ml-1) remained low within the value for normal subjects. These results suggest that some populations of myeloid lineage cells have the ability to produce HGF.     

Clinical significance of arginase in colorectal cancer.

Arginase, a potent immune inhibitor, existed in much greater abundance in the cytoplasm of cancer cells than in normal cells. Serum arginase levels from 31 patients with colorectal adenocarcinoma were determined by using enzyme immunoassay (mean +/- standard error = 18.96 +/- 4.83 ng/ml) and showed to be significantly higher than levels from control subjects (n = 115, 3.09 +/- 0.22 ng/ml) (P less than 0.005). Surgical samples of 15 patients were individually homogenized and assayed by the same method and revealed that the arginase level in tissues with colorectal cancer was two times greater than the level found in normal mucosal tissues (1.74 +/- 0.31 micrograms/g tissue versus 0.77 +/- 0.09 micrograms/g tissue, P less than 0.005). However, the serum arginase levels in patients with colorectal cancer were independent of their carcinoembryonic antigen (CEA) levels (n = 27, arginase 11.81 +/- 1.88 ng/ml, CEA 17.31 +/- 4.24 ng/ml, r = 0.084, P = 0.666). The results suggested that serum arginase level can be a valuable criterion for preoperative evaluation and possibly postoperative follow-up study. It can also combine with CEA determination to intensify the clinical assessment for colorectal cancer.     

Human growth hormone and insulin-like growth factor-1 enhance the proliferation of human leukemic blasts

As the number of long-term survivors of childhood leukemia increases, growth retardation has emerged as a significant complication. Treatment of these children with growth hormone (GH) has been suggested and sporadically implemented. We, therefore, studied the effect of human GH (hGH) and its by-product insulin-like growth factor-1 (IGF-1) on the growth of leukemic cells in vitro. Under serum-free conditions hGH and IGF-1 induced a significant dose-dependent proliferative effect on promyelocytic leukemia (HL60) and Burkitt's lymphoma (Daudi) cell lines. Anti-hGH antibodies negated the stimulatory effect of hGH and anti-IGF-1 serum abrogated the growth-promoting effect enhanced by IGF-1. Similar statistically significant stimulatory properties were found when freshly obtained marrow cells from four of five acute lymphoblastic leukemia (ALL) of childhood and four acute myelogenous leukemia (AML) patients were studied in ALL and AML blast-cell clonogenic assays. ALL colonies increased numerically by 72% (P less than .025) and AML colonies by 92% (P less than .01) in the presence of hGH at concentrations of 2.5 x 10(2) and 3.0 x 10(2) ng/mL, respectively. IGF-1 stimulated ALL and AML blast-colony growth at concentrations ranging from 0.05 to 0.5 ng/mL by up to 105% (P less than .025) and 65% (P less than .03), respectively. Our in vitro data suggest that circulating hGH and IGF-1 may promote leukemic blast cell replication in vivo, and the supplemental administration of hGH to leukemia patients in remission must be carefully monitored for early relapse.     

Death during induction therapy and first remission of acute leukemia in childhood: the St. Jude experience

BACKGROUND: Despite improvements in supportive care, death due to treatment toxicity remains a significant problem for children treated for acute leukemia. METHODS: To determine the causes of and risk factors for death unrelated to refractory leukemia, to disease recurrence, or to second malignancy, the authors reviewed the records of 1011 patients with acute lymphoblastic leukemia (ALL) and 260 patients with acute myeloid leukemia (AML) treated between 1984 and 1999 and between 1983 and 2002, respectively, at St. Jude Children's Research Hospital (Memphis, TN). Data for patients who underwent stem cell transplantation were censored at the time of transplantation. RESULTS: For patients with ALL, the estimated 10-year cumulative incidence of death was 2.9% +/- 5.3%. Age was the only predictor of death. Patients with ALL 1-9 years old had a significantly lower risk of death than did younger or older patients (P = 0.002). For patients with AML, the estimated 5-year cumulative incidence of death was 7.6% +/- 1.9%. Increasing age and increasing leukocyte count were significantly associated with increased risk of death. For patients with ALL and with AML, the incidence of death remained relatively constant during the time periods studied. Infection was the most common cause of death. CONCLUSIONS: In the current study, the authors determined that children > or = 10 years of age are at increased risk of death during therapy for ALL and AML.     

Serum neuron-specific enolase in children with neuroblastoma. Relationship to stage and disease course

Serum neuron-specific enolase (NSE) was measured in 61 children at diagnosis with all stages of neuroblastoma. The median serum values for Stages I, II, III, IV, and IV-S were 13, 23, 40, 214, and 40 ng/ml, respectively. Mean serum levels were different between groups I versus IV, (P = 0.0004) II versus IV (P = 0.0001) and IV-S versus IV (P = 0.004). The prognostic value of serum NSE for disease-free survival was determined in 54 patients at risk for relapse 2 or more years after diagnosis. The disease-free survival rate of all patients with levels of less than 100 ng/ml was 27/34 (79%), whereas it was 2/20 (10%) for those with higher levels. In 28 patients with lower stage disease and a good prognosis (Stages I, II, and IV-S) NSE levels were not predictive of relapse. Only 1 of these 28 patients had a raised level (greater than 100 ng/ml) and survived without relapse, whereas 4 patients who relapsed had serum NSE less than 100 ng/ml at diagnosis. In patients with Stages III and IV disease, a raised serum NSE level was associated with poor outcome: only 1/19 (5%) survived with NSE levels greater than 100 ng/ml, whereas survival was 5/8 (63%) with values below 100 ng/ml. Serial samples were analyzed on 17 patients; all 8 patients with initial NSE levels greater than 100 ng/ml achieved near normal levels during remission (median, 21 ng/ml). However, in only 4/10 patients studied at time of relapse, did the levels rise coincident with relapse. The sera of 47 patients with other forms of cancer and 19 siblings of cancer patients were at or near the normal limits (0-15 ng/ml), with three exceptions: acute lymphoblastic leukemia (286 ng/ml), hepatoblastoma (176 ng/ml), and primitive neuroectodermal tumor (105 ng/ml). Serum NSE is a useful marker for patients with advanced neuroblastoma in whom elevated levels were associated with a poor outcome; the raised NSE levels returned to near normal after therapy. In patients with Stage IV-S disease serum NSE levels were significantly lower than those in Stage IV despite their extensive tumor burden. Serum NSE estimation may confirm Stage IV-S status and suggest a more benign clinical course.     

In vitro activity of vinorelbine on human leukemia cells.

Vinorelbine (VNR) is a semi-synthetic Vinca rosea alkaloid that has been employed both as a single agent and in combination, and has shown significant antitumor activity. As little is known about VNR activity on human leukemia, we studied its in vitro cytotoxic effect on human leukemia cell lines (FLG 29.1, HL60, K562, Balm 4, CEM and Daudi) and on fresh leukemia cells from 28 patients: 2 acute myeloid leukemia (AML); 3 chronic myeloid leukemia in blastic phase (CML-BP); 5 acute lymphoblastic leukemia (ALL); 18 B-chronic lymphatic leukemia (B-CLL), employing the colorimetric INT assay and determining the IC50. We observed that VNR exerts its cytotoxic activity on leukemic cell lines in a dose-dependent fashion. The lymphoid cell lines appear more sensitive than the myeloid ones to the VNR-dependent growth inhibition. A similar pattern was noticed for leukemia cells in primary cultures. VNR is not effective on CML-BP cells, shows variable activity on the AML and ALL cells and is very effective against B-CLL cells. VNR inhibited the growth of fresh B-CLL cells from 15 of 18 patients, the IC50 doses ranging from 4 ng/ml to 83 microg/ml (doses coinciding with the plasma levels obtained in clinics). These observations strongly suggest that VNR could be useful in clinics for the treatment of B-CLL.