中性白细胞弹性蛋白酶与白血病

中性白细胞弹性蛋白酶(NE)是中性自细胞分泌的一种重要细胞毒性分子,与人类白血病的发生密切相关.它能分解早幼粒细胞白血病(PML)-维甲酸受体α(RARα)融合蛋白,增强PML-RABα致白血病的作用,在急性早幼粒细胞白血病发病机制中占有重要地位;另外,它也能通过降低生长因子对正常血细胞生成的有效性,从而促进慢性髓细胞性白血病的克隆优势.     

In vitro induction of myeloid leukemia-specific CD4 and CD8 T cells by CD40 ligand-activated B cells gene modified to express primary granule proteins.

The primary granule proteins (PGP) of myeloid cells are a source of multiple antigens with immunotherapeutic potential for myeloid leukemias. Therefore, we developed a method to induce T-cell responses to PGP protein sequences. We found that gene-transfected antigen-presenting cells efficiently expand functionally competent PGP-specific CD4 and CD8 T cells. The system was optimized using T-cell responses to autologous CD40-activated B cells (CD40-B) transfected with a cytomegalovirus pp65-encoding expression vector. To generate leukemia-specific T cells, expression vectors encoding the PGP proteinase 3 (PR3), human neutrophil elastase, and cathepsin-G were transfected into CD40-B cells to stimulate post-allogeneic stem cell transplantation T cells from five patients with myeloid and three with lymphoid leukemias. T-cell responses to PGP proteinase 3 and human neutrophil elastase were observed in CD8+ and CD4+ T cells only in patients with myeloid leukemias. T-cell responses against cathepsin-G occurred in both myeloid and lymphoblastic leukemias. T cells from a patient with chronic myelogenous leukemia (CML) and from a posttransplant CML patient, expanded against PGP, produced IFN-gamma or were cytotoxic to the patient's CML cells, demonstrating specific antileukemic efficacy. This study emphasizes the clinical potential of PGP for expansion and adoptive transfer of polyclonal leukemia antigen-specific T cells to treat leukemia.     

Breast cancer cell uptake of the inflammatory mediator neutrophil elastase triggers an anticancer adaptive immune response

There is little understanding of the impact of tumor-associated neutrophils (TAN) on adaptive immunity to tumors. In this study, we report the results of an investigation of the pathobiologic basis for the prognostic significance of neutrophil elastase, a serine protease found in neutrophil granules, in a model of cyclin E (CCNE)-overexpressing breast cancer. We established that neutrophil elastase was expressed by TAN within breast cancer tissues but not by breast cancer cells. Neutrophil elastase modulated killing of breast cancer cells by CTLs specific for CCNE-derived HLA-A2-restricted peptide (ILLDWLMEV). Breast cancer cells exhibited striking antigen-specific uptake of neutrophil elastase from the microenvironment that was independent of neutrophil elastase enzymatic activity. Furthermore, neutrophil elastase uptake increased expression of low molecular weight forms of CCNE and enhanced susceptibility to peptide-specific CTL lysis, suggesting that CCNE peptides are naturally presented on breast cancer cells. Taken together, our findings reveal a previously unknown mechanism of antitumor adaptive immunity that links cancer cell uptake of an inflammatory mediator to an effective cytolytic response against an important breast cancer antigen.     

Curcumin inhibits tumor proliferation induced by neutrophil elastase through the upregulation of α1-antitrypsin in lung cancer

Lung carcinogenesis is a complex process in an unregulated inflammatory environment. Curcumin has been extensively investigated as a multi-target anti-tumor and anti-inflammation compound. In this paper, we demonstrate a novel inflammation-related mechanism for curcumin-induced inhibition of lung tumor growth. We found that neutrophil elastase, an important regulator of inflammatory processes, directly triggered tumor cell proliferation in human lung adenocarcinoma A549 cells, and curcumin could completely suppress the excess tumor proliferation induced by neutrophil elastase. α1-antitrypsin is synthesized by tumor cells and is the natural inhibitor of neutrophil elastase. We found that curcumin counteracted the decrease of α1-antitrypsin induced by neutrophil elastase by inducing the promoter activity of α1-antitrypsin and promoting its expression in A549 cells. The inhibition of neutrophil elastase-induced proliferation by curcumin was dependent on the PI3K/Akt pathway. Knockdown of α1-antitrypsin by siRNA further enhanced the tumor cell proliferation induced by neutrophil elastase and significantly blocked the anti-proliferation effect of curcumin against neutrophil elastase. Curcumin remarkably inhibited the primary tumor growth of Lewis lung carcinoma (LLC) in C57BL/6 mice. We further showed that curcumin upregulated the level of α1-antitrypsin in primary tumor tissue by promoting its local expression, and the protein level of neutrophil elastase in tumor tissue was obviously decreased in mice treated with curcumin. Overall, our results suggest that neutrophil elastase and α1-antitrypsin play important roles in modulating lung tumor proliferation in inflammatory microenvironment and curcumin inhibits neutrophil elastase-induced tumor proliferation via upregulating α1-antitrypsin expression in vitro and in vivo.     

Acute lymphoblastic leukemia following severe congenital neutropenia or de novo ALL?

Acute lymphoblastic leukemia (ALL) presenting with neutropenia alone is very rare. We describe a newborn with an early life-threatening infection, severe neutropenia and bone marrow findings compatible with severe congenital neutropenia (SCN). She was treated with granulocyte colony-stimulating factor (G-CSF) with complete neutrophil recovery. Three months later she developed a pro-B ALL. We identified a rare loss of 5′-MLL present at the diagnosis of SCN and ALL by FISH analysis using two different MLL (11q23) probes. Molecular analyses for SCN causing mutations (ELA-2, HAX-1 and G6PC3) and for somatic mutations of the CSF3R gene were negative. The early presence of 5′-MLL loss in bone marrow samples may favor the diagnosis of de novo ALL. Nevertheless, the genetic background for SCN is heterogeneous and a non-described mutation for SCN followed by a secondary ALL cannot be excluded. Further genetic investigation may be useful to gain insight into this rare condition in children.     

Role of imbalance between neutrophil elastase and alpha 1-antitrypsin in cancer development and progression

Neutrophil elastase and alpha 1-antitrypsin are a pair of protease and protease inhibitor counterparts. The imbalance between the two counterparts is generally thought to cause tissue damage, which could create a favourable tissue environment for carcinogens and tumour progression. Laboratory research and clinical findings have indicated that a deficiency in alpha1-antitrypsin is associated with increased risk of liver cancer, bladder cancer, gall bladder cancer, malignant lymphoma, and lung cancer. Conversely, raised concentrations of neutrophil elastase might promote the development, invasion, and metastasis of many cancers. Several mechanisms of carcinogenesis have been postulated. Excess neutrophil elastase might facilitate cancer development by causing tissue damage and air trapping, which foster longer carcinogen exposure, might promote cancer progression by degrading the intercellular matrix barrier, and might directly lead to cancer development through the tumour-necrosis-factor signalling pathway.     

Neutrophil Function in Hematological Neoplasia

Though the main reason for infection in hematologic malignancies remains the reduction in the neutrophil count, there are certain conditions where agranulocytosis alone could not account for this tendency. In non lymphatic disorders, obvious modifications in mature neutrophil functions have been described. They predict susceptibility to infection and bad prognosis in myelodysplastic syndromes (MDS) and in acute non lymphatic leukemia (ANLL) where the mature looking neutrophils represent the leukemic clone and function aberrantly. The defects in neutrophil function encountered in myeloproliferative diseases including chronic myeloid leukemia, are mostly of scientific interest, and usually do not alter the clinical course of the disease in these patients.

The alterations in neutrophil function in the lymphoproliferative disorders result mostly from humoral defects, mainly immunoglobulin deficiency and a variety of neutrophil inhibitory substances, and are not due to primary cellular defects. Neutrophil dysfunction may also result from a protracted infectious course and various treatment modalities, includng splenectomy and chemo-radiotherapy. Neutrophil function studies in splenectomised patients and in selected cases with MDS and ANLL, and studies of serum immunoglobulins and neutrophil inhibitors in patients with lymphoid diseases, may be useful in screening those cases who are prone to infectious problems, and who might therefore benefit from increased infectious precautions.     

Prevalence and impact of colony stimulating factor 3 receptor (CSF3R) mutations among Egyptian acute myeloid leukemia patients

Granulocyte-colony stimulating factor receptor (G-CSFR) mutations have been implicated in the progression of severe congenital neutropenia (SCN) to leukemia. This study aimed to investigate the prevalence of colony stimulating factor 3 receptor (CSF3R) mutations among Egyptian acute myeloid leukemia and their clinic-pathological impact. The study was conducted on 179 adult patients (156 de novo AML and 23 secondary AML on top of SCN). CSF3R mutations were analyzed by sequencing of the PCR products. CSF3R mutations were detected in 2 cases out of 156 de novo AML patients (1.2%) and eighteen cases out of 23 secondary AML patients (78.2%). It was noticed that most of the mutant cases are of younger age, have a high white blood cells count, high bone marrow blasts, bad performance status, and absence of extra medullary disease and with low rate induction remission. Also the overall survival of AML patient's secondary to CSF3R mutations was shorter as compared to those with wild type AML cases. In conclusion the frequency of CSF3R mutations is highly prevalent among AML patients secondary to SCN compared to de novo AML.     

Neutropenia in 6 ethnic groups from the Caribbean and the U.S

BACKGROUND: Low white blood cell counts (WBC) or absolute neutrophil counts (ANC) may delay or prevent the completion of appropriate chemotherapy, especially among women receiving adjuvant therapy for breast and colon cancer, and affect cancer survival. Because race/ethnicity is also associated with survival, the authors compared WBC and ANC in healthy American-born women of African descent and European descent, and women from Barbados/Trinidad-Tobago, the Dominican Republic, Haiti, and Jamaica. METHODS: Blood samples from 261 healthy women ages 20 to 70 years were tested for WBC with differential, cytokine and growth factor levels, and ancestry informative and neutrophil elastase polymorphisms. The authors analyzed the association between neutropenia and serum WBC growth factor levels, cytokine levels, and neutrophil elastase c199a polymorphism. RESULTS: The median WBC and ANC differed among the 6 groups (P < .01 for WBC and P < .0001 for ANC). Dominicans were found to have higher median WBC and ANC than all other groups (P < .03). Neutropenia (ANC < 1500 cu/mm) was observed among 2.7% to 12.5% of the groups of predominantly African descent; no other groups were found to have neutropenia (P < .05). Granulocyte-colony-stimulating factor was found to be lower in white women, but tumor necrosis factor-alpha and C-reactive protein were not found to be correlated with ethnicity. Women of African origin were more likely to have polymorphisms of African ancestry (P < .001) and c199a alleles (P < .0001), which were also associated with low ANC levels. CONCLUSIONS: In the current study, the authors observed a strong association between neutropenia and African descent among asymptomatic women from the U.S. and the Caribbean. Among women of African descent who develop a malignancy, this association may contribute to racial disparities in treatment and outcomes.     

白血病细胞吞噬血细胞现象的观察

对８例白血病细胞吞噬血细胞现象作了较详细的观察描述。被吞噬的主要为红细胞。吞噬后形成假性玫瑰花环样。吞噬红细胞可能是白血病患者贫血原因之一。认为多个白血病细胞围吞一个中性粒细胞的现象值得进一步研究。对吞噬的机理和临床意义作了初步探讨。     

Alpha-1-antitrypsin Phenotypes and Neutrophil Elastase Gene Promoter Polymorphisms in Lung Cancer

Imbalance between neutrophil elastase and alpha-1-antitrypsin (AAT) leads to emphysema in smokers as well as in patients with inherited alpha-1-antitrypsin deficiency. AAT as a proven inhibitor of apoptosis may play role in lung cancer (LC) progression. The aim was to analyse AAT protein variants and polymorphism in promoter region of the neutrophil elastase gene (ELA2) in patients with primary lung cancer. AAT phenotypisation by isoelectric focusing method and ELA2 gene promoter characterization by DNA sequencing were performed in 66 patients with primary lung cancer. Results showed that the frequency of M1 allele and PiM1 homozygotes in LC patients was significantly higher when compared to the healthy subjects (f = 0.6360 and 0.7424 respectively). The most frequent ELA2 promoter region genotypes in LC patients were −903TT and −741GG. There were significantly more patients with intermediate and high ELA2 genotype activity, compared to those with low activity (91% vs. 9%, respectively). In conclusion, we found that PiM1 homozygosity could be associated with the lung cancer, probably due to increased synthesis of this antiapoptotic protein. Non-MM variants of AAT and ELA2 genotypes with predicted intermediate or high activity could also represent a risk factor for aggressive form of lung cancer associated with extrathoracic metastases.     

Clinical evaluation of cefozopran as treatment for febrile neutropenia

Clinical effects and safety of cefozopran (CZOP) were evaluated by the Okayama Bone Marrow Transplantation Group. Twenty-five patients expected to experience febrile neutropenia during induction chemotherapy or consolidation chemotherapy of acute leukemia were enrolled between July 2000 and November 2002. CZOP was administrated by drip infusion at 4g/day bid for a minimum of 3 days. The clinical effects and safety were evaluated in 20 patients with fever of 37.5 degrees C or more from a clinically suspected infection. The underlying disease was acute myeloid leukemia in 17 patients, acute lymphoid leukemia in 1 and acute promyelogeneous leukemia in 1. The complicating infections were sepsis and suspected sepsis. Clinical efficacy was excellent in 11 patients, good in 1, fair in 2 and poor in 6, with an efficacy rate of 60.0%. The efficacy rate in patients whose albumin levels before therapy were less than 3.8 g/dl was 37.5%, whereas the rate in patients whose albumin levels before therapy were between 3.8 g/dl and 5.3 g/dl was 80.0%. The efficacy rate in patients whose neutrophil counts before therapy were less than 100/microliter was 50.0%, whereas the rate in patients whose neutrophil counts after therapy were less than 100/microliter was 53.8%. The efficacy rate in patients whose neutrophil counts both before and after therapy were less than 100/microliter was 37.5%. Side effect of exanthema was observed in 1 patient. These results indicate that CZOP is an effective and safe antibiotic for the treatment of febrile neutropenia in patients with hematological malignancies.     

Empirical examination of the neutrophil criterion (>1500 microl(-1)) currently needed to declare CR in AML

Currently, NCI guidelines require that the neutrophil count exceed 1500 before complete remission (CR) may be declared in acute myeloid leukemia (AML). We tested the empirical validity of this formulation by comparing event-free survival (EFS) in CR in (a). 305 patients who met the NCI criteria for CR and (b). 36 patients who met all the criteria for CR except that the neutrophil count at what we considered CR was 1000-1499 (lower neutrophil group) at which time they began post-remission therapy. EFS was statistically identical in both groups. We extended these findings to 752 patients, who met the NCI criteria for CR except that some may have had circulating blasts at "CR"; 82 of the 752 were in the lower neutrophil group. These data suggest that the minimum needed to declare CR in AML be changed from 1500 to 1000.     

老年人急性白血病患者医院感染危险因素的Logistic回归模型分析

目的 探讨老年人急性白血病(AL)医院感染的特点,分析引起医院感染的危险因素,为医院感染防治措施的制定提供客观依据.方法 对1999年1月至2008年1月9年间住院的116例老年AL患者医院感染发生率、感染部位、致病菌和易感因素等进行回顾性分析,并与同期非老年人组进行比较.结果 老年人组116例中发生医院感染73例(62.9%),感染死亡38例(52.1%),明显高于同期非老年人组,差异有统计学意义(P＜0.01).感染部位以口腔和呼吸道感染为主;多部位感染、重症感染高于非老年人组.致病菌以革兰阴性杆菌为主.多因素分析显示外周血中性粒细胞(ANC)绝对值、化疗周期、近期感染、治疗阶段、住院时间及住院季节是老年人AL医院感染的独立危险因素.结论 老年人AL医院感染发生率高,感染死亡率高,外周血ANC绝对值、化疗周期、近期感染、治疗阶段、住院天数及住院季节是独立危险因素.     

老年人急性白血病患者医院感染危险因素的Logistic回归模型分析

 目的　探讨老年人急性白血病（AL）医院感染的特点,分析引起医院感染的危险因素,为医院感染防治措施的制定提供客观依据。方法　对1999年1月至2008年1月9年间住院的116例老年AL患者医院感染发生率、感染部位、致病菌和易感因素等进行回顾性分析,并与同期非老年人组进行比较。结果　老年人组116例中发生医院感染73例（62.9 %）,感染死亡38例（52.1 %）,明显高于同期非老年人组,差异有统计学意义（P＜0.01）。感染部位以口腔和呼吸道感染为主;多部位感染、重症感染高于非老年人组。致病菌以革兰阴性杆菌为主。多因素分析显示外周血中性粒细胞（ANC）绝对值、化疗周期、近期感染、治疗阶段、住院时间及住院季节是老年人AL医院感染的独立危险因素。结论　老年人AL医院感染发生率高,感染死亡率高,外周血ANC绝对值、化疗周期、近期感染、治疗阶段、住院天数及住院季节是独立危险因素。     

老年人急性白血病患者医院感染危险因素的Logistic回归模型分析

目的 探讨老年人急性白血病(AL)医院感染的特点,分析引起医院感染的危险因素,为医院感染防治措施的制定提供客观依据.方法 对1999年1月至2008年1月9年间住院的116例老年AL患者医院感染发生率、感染部位、致病菌和易感因素等进行回顾性分析,并与同期非老年人组进行比较.结果 老年人组116例中发生医院感染73例(62.9%),感染死亡38例(52.1%),明显高于同期非老年人组,差异有统计学意义(P＜0.01).感染部位以口腔和呼吸道感染为主;多部位感染、重症感染高于非老年人组.致病菌以革兰阴性杆菌为主.多因素分析显示外周血中性粒细胞(ANC)绝对值、化疗周期、近期感染、治疗阶段、住院时间及住院季节是老年人AL医院感染的独立危险因素.结论 老年人AL医院感染发生率高,感染死亡率高,外周血ANC绝对值、化疗周期、近期感染、治疗阶段、住院天数及住院季节是独立危险因素.     

Function of RARalpha during the maturation of neutrophils.

The retinoic acid receptor alpha gene is the target of chromosomal rearrangements in all cases of acute promyelocytic leukemia (APL). This recurrent involvement of RARalpha in the pathogenesis of APL is likely to reflect an important role played by this receptor during the differentiation of immature myeloid cells to neutrophils. RARalpha is a negative regulator of promyelocyte differentiation when not complexed with RA, and stimulates this differentiation when bound to RA. Since RARs are dispensable for the generation of mature neutrophils, their role thus appears to be to modulatory, rather than obligatory, for the control of neutrophil differentiation. In vitro, retinoic acid is also a potent inducer of neutrophil cell fate, suggesting that it might play a role in the commitment of pluripotent hematopoietic progenitors to the neutrophil lineage. Thus, the APL translocations target an important regulator of myeloid cell differentiation.     

慢性中性粒细胞白血病2例并文献复习

目的：研究对慢性中性粒细胞白血病（CNL）的特点。方法：报告2例CNL并结合文献进行复习。结果;患者中性粒细胞明显增多,脾大,中性粒细胞碱性磷酸酶积分增高,无Ph染色体、bcr-abl融合基因,骨髓粒系增生,无病态造血和明显纤维化依据,排除了如类白血病反应、其他克隆性血液病引起的中性粒细胞增多,其中1例伴有单克隆免疫球蛋白血症,2．5年后转化为急性白血病。结论;CNL是一种少见的骨髓增殖性疾病,多发生于老年人,预后差,治疗无标准方案,年轻患者应进行异基因造血干细胞移植以达到治愈目的。