Culture requirements for induction of dendritic cell differentiation in acute myeloid leukemia

Peripheral blood mononuclear cells (PBMCs) from 15 newly diagnosed acute myeloid leukemia (AML) patients were cultured in fetal calf serum-free media supplemented with either granulocyte/macrophage-colony stimulating factor (GM-CSF), interleukin (IL)-4 and tumor necrosis factor α (TNFα), or GM-CSF, stem cell factor (SCF), TNFα and transforming growth factor β (TGFβ) in order to generate leukemia-derived dendritic cells (DCs). Cultured cells were analyzed by flow cytometry with respect to DC-associated surface molecules (CD1a, CD83, CD40, CD80, CD86, HLA-DR) when they showed significant DC morphology in culture (14 cases). After cultivation, neo-expression or upregulation of CD1a antigen was found in 8 samples, CD83 in 2, CD40 in 14, CD80 in 7, and CD86 in 9. Twelve of 14 AMLs, in which DC morphology could be induced upon cultivation, showed upregulation of at least 2 DC-associated molecules. For induction of DC differentiation, GM-CSF, IL-4 plus TNFα was superior in 11 cases, and better results were obtained with GM-CSF, SCF, TNFα plus TGFβ in 3 cases. In 7 of 14 samples tested, a marked increase of the T-cell stimulatory capacity could be demonstrated in the allogeneic mixed lymphocyte reaction. The leukemic origin of in vitro-generated DCs was demonstrated by fluorescence in situ hybridization in a patient with translocation t(15;17). Our results suggest that the use of different culture conditions may extend the number of AML patients in which a differentiation towards the DC lineage can be induced in vitro. Received: 19 August 1999 / Accepted: 13 December 1999     

Myeloperoxidase cytochemical negativity: an unexpected but intrinsic property of blasts of all phases of chronic myeloid leukemia

Myeloperoxidase (MPO) cytochemical activity, recognized as a very important hallmark of myeloblasts, is generally negative in chronic myeloid leukemia (CML) blast crisis (BC). Because this finding is unexpected, being not in keeping with the myeloproliferative nature of CML, we tried to ascertain if MPO cytochemical negativity could be an intrinsic property of blasts of CML and hence present in the preblastic phases as well. Myeloperoxidase cytochemistry of peripheral blood blasts in 161 cases of CML, including 103 in chronic phase (CP) and 29 each in accelerated phase (AP) and BC, was assessed and compared with that of 30 cases of acute myeloid leukemia, AML-M2. Blasts of 97 (94.2%) of 103 cases of CP, 28 (96.6%) of 29 cases of AP, and 22 (75.9%) of 29 cases of BC were negative for MPO (<3% MPO-positive blasts). Compared with the strong MPO positivity, both in terms of intensity and proportion, in the AML-M2 cases, the positivity in the CML cases was generally weak and was seen in a small number of blasts (5–15%), except in one case of BC with 20% positive blasts. Absence or, at times, weak MPO cytochemical activity is an intrinsic property of blasts of all phases of CML, and use of the term myeloblast in CML should be understood to refer to a cell with this property. This also explains why MPO cytochemistry, despite its high reputation as a myeloid-lineage marker, generally does not help in CML BC. CML BC should therefore be considered as a possible diagnosis along with acute lymphoblastic leukemia, AML-M0, AML-M7, etc., in the setting of MPO-negative blasts. Similarity between MPO expression pattern in CML, i.e., negative in blasts and positive in the more mature cells, and that during maturation of normal myeloid series of cells shows the deranged myelopoiesis of CML to be undisturbed at least with respect to MPO expression. There is need for a more comprehensive study of blasts of preblastic phases.     

Clonal Hematopoiesis and risk of Acute Myeloid Leukemia

Acute Myeloid Leukemia, the most common form of acute leukemia in adults, is an aggressive hematopoietic stem cell malignancy that is associated with significant morbidity and mortality. Though AML generally presents de novo, risk factors include exposure to chemotherapy and/or radiation, as well as both familial and acquired bone marrow failure syndromes. Clonal Hematopoiesis (CH) refers to an expansion of blood or marrow cells resulting from somatic mutations in leukemia-associated genes detected in individuals without cytopenias or hematological malignancies. While CH is considered part of normal ageing, CH is also significantly associated with cardiovascular disease, solid tumors, and hematological malignancies. In this review, we will discuss evidence linking CH with the development of AML, as well as describe challenges in and strategies for monitoring patients with high risk CH mutations.     

Treatment of Ph+ chronic myeloid leukemia by gamma interferon

Summary The clinical, hematologic and cytogenetic effects of human recombinant gamma interferon (IFN) were investigated in 14 patients with Ph+ chronic myeloid leukemia (CML). Gamma-IFN was given at a daily dosage of 0.50 mg (=10×10⁶ U)/m² from the 3rd week of treatment on, but the dosage had to be reduced to 0.25 mg/m² in 10 cases and to 0.35 mg/m² in 2 cases, because of the severity and persistence of side effects (mainly fever, fatigue, headache and pain). Only 2 patients tolerated the full dosage. The overall response rate was 64% (1 complete and 8 partial hematologic responses). Only patients in stable chronic phase responded. Two out of two patients in unstable chronic phase and two out of two patients in accelerated phase failed to respond. Eight out of nine responding patients remained in remission throughout the duration of treatment (30 to 35 weeks). No karyotypic conversion was detected. These data show that gamma IFN alone is effective in Ph+ CML, but that side effects can limit substantially the dosage and duration of treatment.     

Imatinib Mesylate in the Treatment of Chronic Myelogenous Leukemia

Imatinib mesylate binds to the inactive conformation of BCR-ABL tyrosine kinase, suppressing the Philadelphia chromosome-positive clone in chronic myelogenous leukemia (CML). Clinical studies of imatinib have yielded impressive results in the treatment of all phases of CML. With the higher rates of complete cytogenetic response with imatinib, molecular monitoring of disease has become mandatory in assessing response and determining prognosis. The practical aspects of the treatment of CML with imatinib are discussed. The emergence of imatinib resistance, albeit in a small percentage of patients, has prompted an evaluation of innovative treatment strategies.     

herg1基因调控慢性髓细胞白血病细胞的增殖和细胞周期

目的:研究herg1基因在慢性髓细胞白血病(CML)细胞的表达及其对CML细胞增殖和细胞周期的调控作用.方法:应用RT-PCR方法测定33例初治慢性髓细胞白血病患者和10例健康志愿者的骨髓细胞herg1基因的表达,检测特异性抑制剂处理后CML细胞株K562增殖凋亡和细胞周期的变化.结果:①在初发CML病例中,herg1 mRNA的表达率是72.7% (24/33),高于正常对照(20%)(P<0.01).CML加速期和急变期的herg1 mRNA的表达率分别为75%和90.9%,高于正常对照(P<0.01),但是herg1的表达与CML的临床分期并无相关性(P>0.05).②E-4031(1 μmol/L)可呈时间依赖性抑制细胞增殖,不诱导明显凋亡.③E-4031阻滞K562细胞周期于G1期.结论:CML细胞herg1基因表达失调,IHERG抑制剂E-4031对K562细胞的增殖和细胞周期具有调控作用,herg1基因可能与CML发病机制相关.     

Systemic inflammatory changes after pulmonary vein radiofrequency ablation do not alter stem cell mobilization

Aims: Aim of this study was to investigate the number of circulatingprogenitor cells, systemic inflammatory mediators, and myocardialnecrosis in patients with paroxysmal atrial fibrillation (AF)undergoing pulmonary vein (PV) isolation by radiofrequency (RF)ablation. Radiofrequency ablation generates a localized myocardialnecrosis that might result in a release of inflammatory mediatorsenhancing progenitor cell mobilization and improving tissuerepair. Methods and results: Blood samples were collected in patients with paroxysmal AFbefore and after PV isolation. Interleukin (IL)-6, IL-1β,TNF-, IL-8, IL-10, and IL-12, and stromal derived factor (SDF)-1were measured by immunoassay. CD34+CD133+, CD117+, and endothelialprogenitor cells (EPCs) were analysed by flow cytometry andculture assay. After ablation procedure, a rise in creatinekinase and troponin T levels indicated myocardial necrosis.Leukocyte counts and C-reactive protein and IL-6 levels increasedsignificantly. Myocardial necrosis and inflammatory responsecorrelated with an increase in IL-6 (P = 0.007). In contrast,SDF-1 levels decreased after RF ablation (P = 0.004). Yet, nosignificant changes were observed in IL-1β, TNF-, IL-8,IL 10, and IL-12 plasma levels or in the number of circulatingCD34+CD133+ and CD117+ progenitor cells, whereas EPCs decreasedby trend. Conclusion: Although PV isolation by RF ablation in patients with paroxysmalAF induces a systemic inflammatory response associated withmyocardial necrosis, no alterations in circulating progenitorcells were observed. Thus, isolated myocardial necrosis maynot be sufficient to account for progenitor cell mobilization.     

Regulation of Hematopoiesis by Chemokine Family Members

Chemokines, originally designated as chemoattractant cytokines, comprise a large family of molecules that have been implicated in a number of different functions mediated through chemokine receptors. Among these functions are regulatory roles in hematopoiesis that encompass effects on the proliferation, survival, and homing/migration of myeloid progenitor cells. This article reviews the field of chemokine regulation of hematopoiesis at the level of myeloid progenitor cells.     

Chronic myelogenous leukemia and exposure to ionizing radiation — a retrospective study of 443 patients

Exposure to ionizing radiations (Rx) has been implicated as a causative factor of chronic myelogenous leukemia (CML). We performed a retrospective study of 443 consecutive CML patients, looking for a history of significant exposure to Rx, and evaluated the clinical and hematological characteristics in order to find any difference between radiation-related CML patients and those with de novo CML. We identified 406 patients without known exposure to mutagens (group I) and 37 patients with prior significant exposure to Rx (group II). In comparison to patients of group I, those of group II showed particular clinical and hematological features: significantly lower incidence of bulky splenomegaly (p<0.05) and hyperleukocytosis (WBC>100×10⁹/l;p<0.05); significantly higher incidence of anemia (Hb<10 g/dl;p<0.01). Patients with radiation-related CML had a significantly better survival than those with de novo CML (median survival 61 months vs 42 months;p<0.05). In conclusion, this study of a large cohort of CML patients indicates that the subgroup of patients with a history of significant exposure to ionizing radiation has particular clinical and hematological features at onset (lower tumor burden, higher frequency of anemia) and a better survival.     

Scleroderma and chronic myeloid leukemia: a sheer coincidence, a consequence of long lasting d-penicillamine therapy or a plausible relationship of both diseases?

Systemic sclerosis is a chronic multisystem disorder of unknown etiology characterized by the involvement of skin and visceral organs caused by an accumulation of collagen. It has been reported that the incidence of solid and hematological malignancy increased in systemic sclerosis. Multiple myeloma and chronic lymphocytic leukemia are the most common hematological malignancies seen in patients with systemic sclerosis. Chronic myeloid leukemia (CML) has only rarely been reported so far. We here report a case with CREST (calcinosis, Raynaud’s phenomenon, esophageal dysfunction, sclerodactly, telangiectasia) who developed CML 7 years after the onset of CREST. Ours is the second case with CML developing after the onset of CREST in the literature. We also briefly discuss the possible tendency to hematological malignancy in systemic sclerosis.     

Therapeutic plateletpheresis in a case of symptomatic thrombocytosis in chronic myeloid leukemia

Extreme thrombocytosis is a frequent feature in myeloproliferative disorders which can predispose a person to thrombotic complications. As opposed to other myeloproliferative disorders, symptomatic thrombocytosis is rare in chronic myeloid leukemia. We describe a second case report of chronic myeloid leukemia (Ph chromosome positive) in a patient in chronic phase on hydroxyurea who presented with sudden onset digital cyanosis of the left hand, giddiness, headache and malaise due to extreme thrombocytosis. A 67% global reduction in the platelet count from 1553 x 10(9)/L to 513 x 10(9)/L after two therapeutic plateletpheresis procedures was seen. There was simultaneous improvement in all symptoms except cyanosis on the tip of the middle finger that progressed to dry gangrene. Dramatic reduction in the platelet count and ablation of symptoms by therapeutic plateletpheresis is an effective therapy and should begin as soon as possible.     

Acute Myelogenous Leukemia Concurrent With Untreated Chronic Lymphocytic Leukemia

We report a case of acute myelogenous leukemia (AML) concurrent with untreated chronic lymphocytic leukemia (CLL). An 84-year-old Japanese man was admitted to the Chihaya Hospital with persistent high-grade fever. Morphologic observation of peripheral blood and bone marrow smears revealed a proliferation of blasts and lymphocytosis with small and mature phenotypes. Immunophenotyping of the blast cells revealed CD13+, CD33+, CD34+, and HLA-DR+, and that of the lymphocytes revealed CD5+, CD19+, CD20+, and lambda+ on the cell surface. The peripheral lymphocytes revealed an IgH gene rearrangement. Chromosome analysis of 20 metaphase cells from bone marrow showed numerous abnormalities, containing +8,+11,+21. The patient's disease was diagnosed as AML with trilineage dysplasia concurrent with CLL. The simultaneous occurrence of AML and CLL is extremely rare but should not be overlooked as a possible underlying cause of lymphocyte abnormalities in AML patients.     

Further evidence for the molecular heterogeneity of chronic myeloid leukemia

Summary Cytogenetic and molecular techniques were performed on samples obtained from 29 patients with chronic myelocytic leukemia (CML); 27 were in the chronic phase and two were in blast crisis. A further five cases were also analyzed, two with atypical CML (aCML), one with chronic neutrophilic leukemia (CNL), and two with juvenile CML (JCML). Most of the cases with typical CML were Philadelphia chromosome (Ph) positive and had a rearrangement within the major breakpoint cluster region (M-bcr). One of these cases was shown to be Ph positive but showed no rearrangement within the M-bcr. Two cases with clinical features typical of CML were Ph negative. One of these showed a rearrangement within the M-bcr, but no rearrangement was demonstrated in the other. Both patients in blast crisis were Ph positive and M-bcr positive. One showed a second Ph. Patients with aCML were Ph negative and had no M-bcr rearrangement. A polymorphism within the M-bcr was found withBglII in one case. No Ph chromosome or M-bcr rearrangement was found in CNL or JCML. These data support the molecular heterogeneity reported in CML.     

Imatinib-induced decompensated heart failure in an elderly patient with chronic myeloid leukemia: case report and literature review

Because it is safe and well tolerated, imatinib is a standard first-line therapy for chronic myeloid leukemia (CML). Although there have been sporadic reports of imatinib-induced cardiotoxicity, including left ventricle (LV) dysfunction and heart failure, the evidence for it is contradictory. Here, we reported a case of an 88-year-old male patient with CML developed decompensated heart failure following imatinib therapy. Four days after the initiation of imatinib, the patient developed orthopnea, edema and a pleural effusion accompanied by abdominal distension, nausea and vomiting. The chest X-ray film showed an enlarged cardiac profile. The echocardiogram demonstrated a decreased LV ejection fraction and enlarged left-side cardiac chambers. B-type natriuretic peptide concentrations were markedly increased. The patient recovered soon after the withdrawal of imatinib and introduction of comprehensive therapy for heart failure. Imatinib-induced cardiotoxicity in elderly patients is a potentially serious complication that merits further evaluation.     

Analysis of altered proteins related to blast crisis in chronic myeloid leukemia by proteomic study

Introduction: Chromic myeloid leukemia (CML) blast crisis (BC) and imatinib (IM) resistance is a significant barrier to the effective treatment of the disease. Methods: Expression profiles of differential proteins were identified, and new biomarkers or pathways related to BC in CML were screened through proteomic analysis. Total proteins from primary bone marrow cells of CML patients in chronic phase (CP) and BC were separated via two‐dimensional (2D) polyacrylamide gel electrophoresis and then analyzed by imagemaster 5.0 software to detect differential protein spots which were already identified by mass spectrometry. Based on the variation of the whole expression profile, some key proteins were picked out for Western blot to confirm the accuracy of proteomics data. Moreover, related signal pathways involving those proteins were investigated. Results: The result indicated that thirteen protein points between CML‐CP and CML‐BC were successfully determined. Results from Western blot of RhoA, hnRNPK, ANXA1, PSMB4, and LTA4H were similar to those from 2D polyacrylamide gel electrophoresis. Most of those proteins were involved in the proteosome pathway and the small G‐protein pathway. Conclusion: A group of proteins associated with BC can be obtained and the result of this study might provide clues for further research.     

A study of the myeloid differentiation antigens of the peripheral blood neutrophil granulocytes in the different evolutive phases of chronic myeloid leukemia

Summary To ascertain whether progression from the chronic to the accelerated and blastic phases of chronic myeloid leukemia (CML) is associated with the loss of the myeloid differentiation antigens of the neutrophil granulocytes (NG), we analyzed two monoclonal antibodies (PMN 31D8 and PMN 13F6) recognizing normal peripheral blood NG membrane antigens in 49 patients in different evolutive stages of CML. Since five patients were studied twice, a total of 54 studies were carried out. Fourteen patients were evaluated at diagnosis, 12 in the controlled chronic phase within 1 year from diagnosis, 14 in the advanced chronic phase (median evolution 3.25 years), and 14 in the accelerated (five cases) or blastic (nine cases) phases. Fourteen normal subjects served as a control group. At diagnosis, a significant decrease in the positivity for both antigens was observed with respect to controls, probably due to the circulation of incompletely mature NG. In the early chronic phase the values were within the normal range, whereas a significant decrease was registered in the advanced chronic phase and especially in the accelerated/blastic phase. A negative correlation between the NG positivity for both markers and the time elapsed from the moment of obtaining the initial control of the disease was found, suggesting that a progressive loss of the myeloid antigens of the NG occurs during the evolutive course of CML. These results seem to confirm the usefulness of the NG myeloid differentiation antigen study as an evolutive parameter in CML.This work has been supported by the grant no. 89/0351 fromFondo de Investigaciones Sanitarias de la Seguridad Social M. Rozman is recipient of the grant Beca Hospital Clínic i Provincial 1990     

Endometrial cancer due to busulfan therapy

Summary Two patients with CML who developed endometrial cancer following 2 years of busulfan treatment are reported. The possibility of a causal relationship between the cytotoxic drug and the development of this malignancy is discussed.Tübitak The Unit Hemolytic Anemias and Chemical Hematotoxic Disorders