杀伤细胞KIR与造血干细胞移植

NK细胞的杀伤细胞免疫球蛋白样受体(killercellimmunoglobulin-likereceptor,KIR)是属于免疫球蛋白样超家族的一系列分子,表达于自然杀伤(naturekiller,NK)细胞和部分T细胞的表面,能识别HLAI类分子。它们可通过与靶细胞表面的HLAI类分子结合,传导激活或抑制信号,从而调节NK细胞和T细胞的活性。造血干细胞移植(hematopoieticstemcelltransplantation,HSCT)过程中,当供者KIR与受者HLAI类分子不匹配时,供者NK细胞功能不被抑制,从而对受者抗原提呈细胞(antigen-presentingcell,APC)和残留肿瘤细胞产生较强的异源反应。研究也已证实供者NK细胞的异源反应活性可提高移植物植入率、促进移植物抗白血病细胞(graft-versus-leukemia,GvL)作用并降低移植物抗宿主病(graft-versus-hostdisease,GvHD)。     

细胞凋亡与急性淋巴细胞白血病治疗的研究进展

近年来,由于对细胞凋亡研究的深入,急性淋巴细胞白血病的临床治疗也有了新的可能。通过选择性诱导白血病干细胞的凋亡,可对急性淋巴细胞白血病的治疗开辟新的方向。而观察病程中细胞凋亡和其相关因子的情况,也可对急性淋巴细胞白血病的预后起到预报作用。虽然此类治疗方法并未大批量应用于临床,但相信随着细胞凋亡机理的进一步研究和其他工具学科的发展,细胞凋亡的选择性诱导也终将成为治疗急性淋巴细胞白血病的有效途径。因此,本文将对细胞凋亡和急性淋巴细胞白血病治疗的研究进展做一综述。     

南方汉族KIR-HLA系统分子遗传多态性与急性白血病的相关性

目的:探讨中国南方汉族人群KIR-HLA系统分子遗传多态性与急性淋巴细胞白血病(acute lymphoblastic leukemia, ALL)、急性髓系白血病(acute myelocytic leukemia, AML)的相关性。方法:对323份成年ALL、350份成年AML患者以及745份随机健康对照的样本,...     

miRNA-93在急性淋巴细胞白血病中的表达

目的:探讨微小RNA(miRNA)-93在急性淋巴细胞白血病中的表达情况及其对急性T细胞白血病细胞株Jurkat增殖的影响和潜在作用机制。方法:Real-time PCR技术检测急性白血病患者骨髓样本中miRNA-93的表达水平。转染miRNA-93 inhibitor下调Jurkat细胞中miRNA-93的表达,分别采用CCK-8法、Ed U法和流式细胞术检测细胞活力、增殖及周期,Western blot检测周期相关调控因子cyclin D1、细胞周期蛋白依赖性激酶4(CDK4)、p-Rb及P27的蛋白表达水平。结果:miRNA-93在急性白血病患者中呈现高表达,且在高危患者中表达水平最高;沉默miRNA-93后,Jurkat细胞的活力下降并出现明显的G1/S期阻滞,同时细胞中cyclin D1、CDK4、p-Rb的蛋白水平显著降低,而P27的蛋白水平显著升高。结论:miRNA-93在急性淋巴细胞白血病中表达显著升高;沉默miRNA-93可通过调控周期相关因子的表达抑制急性T细胞白血病细胞株Jurkat的增殖。     

KIR及HLA-A、B基因多态性与急性淋巴细胞白血病的相关性

目的探讨急性淋巴细胞白血病患者的杀伤细胞免疫球蛋白样受体（killer cell immunoglobulin-like receptors,KIR）和人类白细胞抗原（human leukocyte antigen,HLA）HLA-A、B等位基因多态性。方法采用Luminex流式技术-序列特异性寡核苷酸探针反向杂交（flow cytometry-sequence specific oligonucleotide probe,FLOW-SSOP）方法对内蒙地区48例急性淋巴细胞白血病患者HLA-A、B等位基因多态性进行分析,PCR-SSP技术进行KIR抑制基因的低分辨率检测。以北方地区健康群体资料作为正常对照。结果 （1）在急性淋巴细胞白血病中HLA-A＊31XX,A＊6901等位基因频率高于对照组（1.955%,0.071%）,差异有统计学意义（P〈0.05）;A＊33XX等位基因频率低于对照组（5.825%）,差异有统计意义（P〈0.05）;（2）在HLA-B等位基因中,急性淋巴细胞白血病B＊07XX,＊27XX,＊38XX,＊41XX,＊49XX,＊59XX等位基因频率高于对照组（2.069%,0.968%,0.702%,0.091%,0.051%,0.061%）,差异有统计学意义（P〈0.05）;其中HLA-B＊59XX的相对危险度高（RR=35.156）,其它等位基因频率在两组之间无显著性差异。在急性白血病患者中,高频率KIR基因有KIR3DP1,2DP1,2DL1,2DL3,3DL1、2DL5,2DS4及3DS1。急性白血病组3DL1基因型频率比对照组的显著降低。结论 HLA-A＊31XX,A＊6901;B＊07XX,B＊27XX,B＊38XX,B＊41XX,B＊49XX,B＊59XX;等位基因与ALL相关联,有遗传易感作用。HLA-B＊59XX等位基因携带者,可能患ALL的危险度增高（RR=35.156）KIR3DL1和2DS4均与ALL呈关联。     

CD4+CD25+Treg在急性淋巴细胞白血病患者外周血中的表达及其体外实验研究

目的初步探讨CD4 CD25 调节性T细胞(CD4 CD25 regulatory T cells,CD4 CD25 Treg)在急性淋巴细胞白血病(acute lymphocytic leukemia,ALL)患者化疗前及化疗缓解后外周血中的表达水平,并研究患者血清能否诱导外周血CD4 CD25-T细胞转化为CD4 CD25 Treg。方法①采用流式细胞术分别检测ALL初诊组、化疗完全缓解或部分缓解组及正常对照组外周血中CD4 CD25 T细胞所占比例,然后通过荧光定量RT-PCR检测各组外周血中转录因子Foxp3mRNA的表达水平,并逐层分析比较。②采集正常人外周血单个核细胞后,对照组用正常人血清,实验组用患者血清并分别设浓度梯度进行培养,72h后采用流式细胞术、荧光定量RT-PCR分别检测CD4 CD25 T细胞和Foxp3mRNA表达。结果ALL化疗缓解组CD4 CD25 T细胞及Foxp3mRNA表达水平均明显高于ALL初诊组和正常对照组(P<0.05),后两者之间CD4 CD25 T细胞水平无统计学差异(P>0.05),但ALL初诊组Foxp3mRNA含量较正常对照组明显升高(P<0.01),差异具有统计学意义;并且血清培养对照组CD4 CD25 T细胞水平及Foxp3mRNA含量均明显低于实验组(P<0.05),且其表达并不随血清浓度的增加而升高。结论CD4 CD25 Foxp3 Treg在ALL初诊组及化疗缓解组患者外周血中比例明显升高,且初步表明患者血清中的可溶性物质可诱导外周血CD4 CD25 T细胞转化为CD4 CD25 Treg,提示CD4 CD25 Treg可能是ALL免疫抑制的一个重要原因。     

KIR在NK-92MI及K562细胞中的基因型及表达谱分析

目的:分析杀伤细胞免疫球蛋白样受体(KIR)在NK-92MI及K562细胞中的基因型及表达谱, 探索KIR受体的表达调控规律.方法:采用PCR、 RT-PCR及分子克隆测序技术, 检测K562及NK-92MI细胞中KIR2DL1、 2DL2、 2DL3、 2DL4、 2DL5、 2DS1、 2DS2、 2DS3、 2DS4、 2DS5、 3DL1、 3DL2、 3DL3、 3DS1、 2DP1、 3DP1的基因型, 及KIR2DL1、 KIR2DL2、 KIR2DL3、 KIR3DL1、 KIR2DS1、 KIR2DS3及KIR2DS2/4受体mRNA的表达.结果:K562细胞携带KIR2DL1、 2DL2、 2DL3、 2DL4、 2DL5A、 2DS2、 2DS4*003-006、 2DS5、 3DL1、 3DL2、 3DL3基因, 但不表达KIR.NK-92MI细胞携带KIR2DL1、 2DL2、 2DL3、 2DL4、 2DS2、 2DS4*001/002、 2DS4*003-006、 3DL1、 3DL2、 3DL3, 但只表达KIR2DL1、 KIR2DL3及KIR2DS2/4.结论:K562细胞及NK-92MI细胞均携带KIR2DL1、 2DL2、 2DL3、 2DL4、 2DS2、 2DS4*003-006、 3DL1、 3DL2、 3DL3.NK-92MI细胞表达KIR受体, K562细胞不表达上述KIR, KIR呈细胞特异性表达.     

91例急性非淋巴细胞白血病的细胞遗传学研究

为探讨急性非淋巴细胞白血病患者细胞遗传学改变,对９１ 例急性非淋巴细胞白血病（ＡＮＬＬ） 的骨髓染色体进行了检查,对染色体畸变,特别是特异性染色体畸变以及染色体畸变发生率作了研究,结果有６７．０％ 的患者有染色体异常改变,其中结构异常占４３．９ ％ ,数目异常占２３ ．１％ 。Ｍ２、Ｍ３、Ｍ５、Ｍ４ 畸变比率最高,共占染色体畸变总数的９０ ．２ ％ ,Ｍ１、Ｍ７ 次之,Ｍ６ 较少有染色体畸变,此畸变与ＡＮＬＬ的细胞类型常带有一种特异的对应关系。同时对不同时期的染色体核型作了动态观察,表明染色体核型动态检查对于观察ＡＮＬＬ的病情及指导治疗和判断预后都有一定的价值。并对ＡＮＬＬ 的细胞遗传学与ＦＡＢ分型、免疫分型间的关系作了探讨,表明形态学、免疫学、细胞遗传学对ＡＮＬＬ分型是不可分割的一个整体。     

白血病患者及其HLA-A/B/DR全相合同胞中杀伤细胞免疫球蛋白样受体基因型分布

目的 通过研究白血病患者及其HLA-A/B/DR相同的同胞中杀伤细胞免疫球蛋白样受体(KIR)基因型的分布情况,探讨同胞间KIR基因型与HLA分子之间的关系及KIR在不同种类白血病中的分布.方法 采用序列特异性弓I物聚合酶链反应(PCR-SSP)法进行KIR基因分型,对78例白血病患者及其HLA-A/B/DR相同的83例同胞的KIR基因型进行分析.结果 78例患者中有48.72%与其同胞有相同的KIR基因型,44.87%的患者与其同胞不完全相同.所有KIR基因型频率在患者及同胞之间的差异均无统计学意义(P＞0.05).除KIR2DS4在慢性粒细胞白血病患者中的频率明显高于急性淋巴细胞白血病和急性非淋巴细胞白血病外,其余KIR基因型在3组疾病中的频率差异均无统计学意义(P＞0.05).结论 KIR基因和HLA Ⅰ类抗原各自独立遗传,且相对稳定.KIR基因受白血病类型影响较小.     

杀伤细胞免疫球蛋白样受体(KIR)基因多态性与系统性红斑狼疮的关联性研究

目的 探讨杀伤细胞免疫球蛋白样受体（KIR）基因多态性与系统性红斑狼疮（SLE）的关联性。方法 采用序列特异性引物聚合酶链反应（SSP-PCR）法，分析93例SLE患者和123例无血缘关系的健康对照K／R基因位点的多态性。结果 SLE病例组KIR2DS1（P〈0．001）、KIR2DL2（P〈0．001）基因的阳性率较随机对照组显著升高。具有2个或2个以上活化性基因个体在SLE组（80．7％）较对照组（66．7％）明显增多，差异具有统计学意义（P=0．025）。SLE患者狼疮肾炎与非狼疮肾炎组K／R基因分布频率比较差异无统计学意义。按发病年龄分组后，SLE患者中不同发病年龄组间K／R基因频率分布比较差异无统计学意义。结论 KIR2DS1、KIR2DL2基因频率升高可能与SLE发病相关。     

Pathobiology of acute lymphoblastic leukemia

In the present review, the authors described the pathobiological features of B- and T-ALL, which appear to be quite heterogeneous with regard to molecular pathogenesis. The last edition of the World Health Organization Classification considered this aspect by defining many entities based on genetic findings. This approach is not only important for prognostic stratification, but also in the near future will surely represent the basis for the definition of patient-specific therapeutic approaches. A striking example is Ph+ acute lymphoblastic leukemia (ALL), which until the advent of tyrosine kinase inhibitors (TKI) has been regarded as the most aggressive ALL. The use of imatinib, dasatinib, and possibly more recent inhibitors has dramatically changed the clinical scenario, offering new opportunities to patients, especially the elderly. Similarly, the use of FLT3 inhibitors in mixed lineage leukemia-positive cases, gamma-secretase inhibitors in T-ALL, novel TKI, and monoclonal antibodies may represent a successful approach in the future.     

KIR Donor Selection: Feasibility in Identifying better Donors

We previously reported that acute myelogenous leukemia (AML) transplants using killer cell immunoglobulin-type receptor (KIR) B haplotype better or best (≥2 B activating gene loci ± Cen B/B) unrelated donors (URDs) yield less relapse and better survival. In this prospective trial we evaluated 535 AML searches from 14 participating centers with centralized donor KIR genotyping for donor selection. This represented 3% to 48% of all AML searches (median 20%) per center, totaling 3 to 172 patients (median 22) per center. Donor KIR genotype was reported at a median of 14 days after request (≤26 days for 76% of searches). In 535 searches, 2080 donors were requested for KIR genotyping (mean 4.3 per search); and a median of 1.8 (range, 0 to 4.5) per search were KIR typed. Choosing more donors for confirmatory HLA and KIR haplotype identification enriched the likelihood of finding KIR better or best donors. The search process identified a mean of 30% KIR better or best donors; the success ranged from 24% to 38% in the 11 centers enrolling ≥8 patients. More donors requested for KIR genotyping increased the likelihood of identifying KIR better or best haplotype donors. Of the 247 transplants, 9.3% used KIR best, 19% used KIR better, and 48% used KIR neutral donors while 24% used a non–KIR-tested donor. KIR genotyping did not delay transplantation. The time from search to transplant was identical for transplants using a KIR-genotyped versus a non–KIR-genotyped donor. Prospective evaluation can rapidly identify KIR favorable genotype donors, but choosing more donors per search would substantially increase the likelihood of having a KIR best or better donor available for transplantation. Transplant centers and donor registries must both commit extra effort to incorporate new characteristics (beyond HLA, age, and parity) into improved donor selection. Deliberate efforts to present additional genetic factors for donor selection will require novel procedures.     

急性淋巴细胞性白血病微小残留病变筛选指标的特点分析

目的 对儿童急性淋巴细胞性白血病微小残留病变(MRD)筛选指标进行分析并评估其意义和表达特点.方法 分离35例初发B-急性淋巴细胞性白血病(ALL)患儿的单个核细胞,对符合CD38、CD45弱表达,CD58、CD21、CD22强表达,CD34和Cu同时表达,染色体相关抗原CD66c表达的,与CD10/CD34/CD19进行四色抗体组合,应用流式仪检测,如在双参数点图上所选择的四色抗体组合出现的位置明显有别于正常骨髓相应位置的,则认定该抗体组合为有效的筛选标记并进行随后的MRD监测.结果 35例患儿中31例存在至少一个MRD标记,覆盖率为88.6%;21/35例患儿(60%)存在2个或2个以上的筛选标记;TdT/CD10/CD34/CD19为最常见的四色组合.结论 TdT/CD10/CD34/CD19作为四色MRD筛选标记覆盖率高,应作为常规和首选的筛选标记;免疫表型中Pro-B缺乏有效的筛选标记,出现2个或以上的筛选标记,对提高MRD的精确度具有重要意义.     

217例成人急性淋巴细胞白血病遗传学特征及其临床预后意义的研究

目的 研究成人急性淋巴细胞白血病(acute lymphoblastic leukemia,ALL)的细胞和分子遗传学特征及其临床预后意义.方法 通过常规细胞遗传学、荧光原位杂交、实时定量多聚酶链反应、巢式PCR扩增及测序等技术相结合,对217例成人ALL患者细胞及分子遗传学特征全面分析,并探讨其临床预后意义.结果 成人ALL遗传学异常以t(9;22)(q34;q11)/ BCR-ABL融合基因最常见(56.3％),其次为复杂核型(13.8％),其它异常占29.9％;经荧光原位杂交确诊BCR/ABL融合基因阳性患者中22.4％经常规细胞遗传学检测未发现Ph染色体;遗传学预后分组高危组患者随年龄增大递增;高危组B-ALL亚型高于标危组(98.4％ vs.65.7％,P=0.000),高危组3个月(67.3％ vs.85.1％,P=0.042)、6个月(55.1％vs.80.4％,P=0.008)、12个月(34.0％ vs.59.1％,P=0.017)和24个月(13.0％ vs.36.6％,P=0.010)总生存(overall survival,OS)率均显著低于标危组,中位OS时间高危组(11个月,95％CI:8.0～13.9)比标危组(19个月,95％CI:10.8～27.1)显著缩短(P=0.001).结论 成人ALL具有特征性遗传学异常,本组预后分组对判断患者预后、指导临床正确分层治疗具有一定临床意义.临床上单独应用常规细胞遗传学检测漏诊率高,建议常规细胞遗传学检测、荧光原位杂交技术和分子生物学技术相结合,提高细胞及分子遗传学亚型的正确诊断水平.     

24小时持续水化、碱化在大剂量氨甲喋呤治疗儿童急性淋巴细胞白血病中的应用探讨

目的进一步探讨具有相同治疗效果但可以降低大剂量氨甲喋呤治疗小儿急性淋巴细胞白血病（ALL）毒副作用的方法。方法对2003年1月至2005年12月收治儿童ALL共31例次HD-MTX治疗采用双通路24h持续水化、碱化为观察组,1998年10月~2002年12月收治儿童ALL共25例次HD-MTX治疗采用全国标准方案为治疗组,观察两组毒副作用的发生率和严重程度以及治疗效果。结果两组毒副作用的发生率和严重程度有显著性差异。结论观察组采用双通路24h持续水化、碱化的毒副作用的发生率和严重程度均明显低于治疗组,值得在基层医院应用及推广。     

Pyomyositis in the course of acute lymphoblastic leukemia

Pyomyositis is a term used to asses pyogenic infection of the skeletal muscle and develops as the result of bacteriemia and occurs most commonly in patients with various immunosuppressive diseases. Pyomyositis should be considered in the differential diagnosis in patients complaining of intensive muscle pain and fever. The usual causative organism is Staphylococcus aureus. We present a 3-year-old boy with acute lymphoblastic leukemia (ALL) and pyomyositis caused by Pseudomonas aeruginosa diagnosed in the course of induction therapy. The diagnosis of pyomyositis in the crural muscles of both legs was given based on imaging examinations: ultrasonography and magnetic resonance.     

Posaconazole and fluconazole prophylaxis during induction therapy for pediatric acute lymphoblastic leukemia

PurposeTo assess and compare the efficacy and safety of posaconazole with fluconazole for the prevention of invasive fungal infections in children who were undergoing induction therapy for acute lymphoblastic leukemia (ALL). To develop an approach to predict invasive fungal infections in ALL patients who accepted posaconazole prophylaxis.MethodsThis was a single-center, retrospective cohort study of patients with newly diagnosed ALL, comparing invasive fungal infections in patients who received no prophylaxis, posaconazole prophylaxis, or fluconazole prophylaxis during induction therapy. A propensity score-weighted logistic regression model was used to adjust for confounders. Hepatotoxicity was assessed according to the Common Terminology Criteria for Adverse Events (CTCAE) criteria.ResultsOut of the 155 ALL patients, 60 received no prophylaxis, 70 received posaconazole prophylaxis, and 25 received fluconazole prophylaxis. Posaconazole prophylaxis reduced the odds of invasive fungal infections by > 60%, prolonged infection-free survival significantly, and did not increase the risk of hepatotoxicity. Additionally, we found that the combination of age at diagnosis, clinically documented bacterial infection in the first 15 days of induction therapy, and absolute neutrophil count (ANC) curve enabled significant prediction of the susceptibility to infections after receiving posaconazole prophylaxis.ConclusionsOur findings supported using targeted prophylaxis with posaconazole in ALL children undergoing induction chemotherapy. Age, clinically documented bacterial infection and ANC are important predictors of invasive fungal infections in patients with posaconazole prophylaxis.     

Ultrastructural myeloperoxidase activity and expression of myeloid-associated antigens in acute lymphoblastic leukemia

Ultrastructural myeloperoxidase (MPO) activity and myeloid-associated antigen (MyAg) expression were investigated in 12 adult patients with acute lymphoblastic leukemia (ALL). Ultrastructural MPO was detected by 3 different methods. Immunophenotyping was performed by flow cytometry, using a series of monoclonal antibodies. Ultrastructural MPO-positive blast cells were detected in 6 patients (50%). In 5 of these 6 patients, the methods detecting both MPO and platelet peroxidase (PPO) activities found MPO-positive blast cells more frequently than those detecting MPO activity alone. In 2 patients (17%), at least one kind of MyAg was positive. Ultrastructural MPO activity was detected more frequently than MyAg expression in ALL patients. This method of detecting PPO and MPO is recommended for detection of ultrastructural MPO-positive ALL.     

Genetic polymorphisms associated with adverse events and elimination of methotrexate in childhood acute lymphoblastic leukemia and malignant lymphoma

Methotrexate is administered in high doses to treat childhood acute lymphoblastic leukemia and malignant lymphoma. Hepatotoxicity and bone marrow suppression often limit its use, however. The objective of this study was to determine the genetic polymorphisms associated with the hepatotoxicity and elimination of methotrexate. Genetic polymorphisms of glutathione S-transferase (GST) genes including GSTT1 positive/null, GSTM1 positive/null, and GSTP1 A313G, and genes for reduced folate carrier 1 G80A (RFC1 G80A), methylenetetrahydrofolate reductase C677T (MTHFR C677T), and breast cancer resistant protein C421A (BCRP C421A) were determined for 26 patients by the polymerase chain reaction (PCR) method or by direct sequencing. A high frequency of hepatotoxicity (P = 0.035) was observed for patients with GSTM1 positive and RFC1 AA⁸⁰, and serum concentrations of methotrexate 48 h after the start of infusion were higher for patients with the TT⁶⁷⁷ genotype of MTHFR (P = 0.028). In conclusion, GSTM1 positive/null and RFC1 G80A polymorphisms could be predictors for hepatotoxicity, and the MTHFR C677T polymorphism is associated with elimination of methotrexate.