系统性红斑狼疮患者B淋巴细胞刺激因子及其受体表达水平的研究

目的：研究系统性红斑狼疮（SLE）患者B淋巴细胞刺激因子（BLyS）及其受体的表达水平，并探讨其与系统性红斑狼疮疾病活动指数（SLEDAI）的关系。方法：检测缓解组（14例）和活动组SLE患者组（23例）血清BLyS及其受体的表达水平，以健康志愿者（30例）作为对照。结果：SLE患者BLyS、穿膜蛋白活化物（TACI）和B细胞活化因子受体（BAFF—R）mRNA的表达明显高于正常人（P〈0．01），活动期患者的表达高于缓解期（P〈0．01）。结论：SLE患者BLyS、TACI和BAFF—R mRNA表达含量升高，提示BLyS、TACI和BAFF—R可能与SLE的发病有关。     

多发性骨髓瘤患者B淋巴细胞刺激因子及其受体的表达水平

目的 探讨B细胞激活因子(BLyS) 及其受体在多发性骨髓瘤(MM)发病中的作用机制.方法 用酶联免疫吸附测定法(ELISA)和实时荧光定量聚合酶链反应(RFQ-PCR),对31例MM患者和30名健康人血清BLyS含量、外周血单个核细胞(PBMCs) 中BLyS及其受体mRNA含量进行检测;并比较MM患者血清BLyS含量与外周血PBMCs中BLyS及其受体基因表达水平和β2微球蛋白(β2M)的相关性.结果 MM患者血清BLyS含量(6.99±3.28 g/L)明显高于健康对照组(2.70±1.09 g/L,P<0.01),并与β2M浓度呈正相关性(r=0.954,P=0.000);77.42%(24/31),93.55%(29/31),90.32%(28/31)的MM患者血清BLyS蛋白和PBMCs中BLyS,B细胞成熟抗原(BCMA) mRNA表达水平较正常人明显增高,并且也与β2M的含量呈正相关;而64.52%(20/31)患者穿膜蛋白活化物(TACI) mRNA表达水平降低.结论 MM患者的血清BLyS蛋白浓度和PBMCs中BLyS及其受体基因表达水平有异常改变,说明BLyS及其受体可能参与MM的发病;BLyS及其受体表达水平也许可作为预后的一个指标.     

增殖诱导配体及其受体在肿瘤组织中的表达与意义

目的分析增殖诱导配体（APRIL）及其受体在不同肿瘤组织及肿瘤发展不同阶段的表达水平,探讨其与肿瘤发生的关系。方法采用实时荧光定量聚合酶链反应（RFQ-PCR）检测了临床上常见肿瘤组织中APRIL及其受体mRNA的准确含量,并以靶基因和内参mRNA含量的比值作为评价靶基因表达水平的指标。结果肿瘤组织中APRIL表达水平显著高于交界组织（P〈0．05）和正常组织（P〈0．05）;B细胞成熟抗原（BCMA）和穿膜蛋白活化物（TACI）表达水平在大多数肿瘤组织中与正常组织差异无统计学意义（P〉0．05）;APRIL在肠上皮化生、异型增生和胃癌组织中表达水平显著高于正常黏膜（P〈0．05）,APRIL在胃癌组织中表达水平显著高于肠上皮化生和异型增生（P〈0．05）,而其受体BCMA和TACI在各组胃黏膜病变之间表达水平无统计学意义（P〉0．05）。结论成功建立RFQ-PCR检测APRIL及其受体mRNA含量的方法,具有较好的检测灵敏度和重复性;APRIL在肿瘤发生、发展过程中可能起了重要作用;除了BCMA和TACI外,肿瘤细胞可能还存在APRIL未知受体;APRIL在某些类型肿瘤有可能主要是通过受体BCMA而不是TACI发挥作用。     

B细胞激活因子在免疫调节中的作用

B细胞激活因子（BAFF）是调节B淋巴细胞存活和成熟的细胞因子,属于TNF家族成员.BAFF有三个受体,分别为BCMA,TACI和BAFF-R.近几年来的研究表明,BAFF及其受体在抗体类型转换、生发中心维持、T细胞共刺激等方面发挥着重要的免疫调节作用,对自身免疫疾病、淋巴瘤和B细胞免疫缺陷的治疗具有潜在的应用前景.本文就BAFF的结构与表达,BAFF受体,生物学功能及其临床应用作一综述.     

B淋巴细胞刺激因子及其受体在多发性骨髓瘤中的表达及其意义

目的 探讨B淋巴细胞刺激因子(BLyS)及其受体在多发性骨髓瘤(MM)中的表达及其意义.方法 采用流式细胞术(FCM)分析人MM肿瘤细胞系KM3及CZ-1细胞表面BLyS及其受体的表达;RT-PCR及Western blot进一步验证Blys及其受体的表达;采用荧光免疫细胞化学法和激光共聚焦技术鉴定KM3细胞中Blys蛋白的表达与定位;WST细胞增殖实验检测Blys对MM肿瘤细胞生长与存活的影响.ELISA及荧光定量.聚合酶链反应(RTQ-PCR)测定MM患者BLyS蛋白与mRNA的表达水平.同时分析乳酸脱氢酶(LDH)、β2微球蛋白浓度与BLyS蛋白及mRNA表达水平的关系.结果 ①MM细胞能够表达BLyS及其受体;②BLyS定位表达于KM3细胞的浆膜上;③Blys促进MM细胞的生长与存活;④MM患者BLyS蛋白或mRNA表达水平显著高于正常对照组(P值均<0.01);⑤直线相关分析显示LDH、β2微球蛋白浓度与BLyS蛋白及mRNA表达水平呈显著相关性.结论 BLyS及其受体对于MM发生、发展可能起着重要的作用.     

JNK信号通路活化与BCMA表达对MM细胞的作用研究

目的研究多发性骨髓瘤(MM)细胞中B细胞活化因子(BAFF)及B细胞成熟抗原(BCMA)的表达及作用,JNK信号通路的表达及活化情况,探讨JNK信号通路活化与BCMA表达间的相互作用。方法采用实时荧光定量(QRT)-聚合酶链式反应(PCR)检测MM患者及健康对照者单核细胞(PBMCs)中BAFF及其受体BCMA、TACI、BAFF-R的表达。采用Western Blot方法检测MM细胞中BCMA蛋白的表达及信号通路蛋白ERK、p-ERK、JNK、p-JNK、p38及p-p38的表达及活化情况;采用WST-1法检测MM细胞的增殖与存活。结果 MM患者PBMCs中BAFF及BCMA的表达显著高于健康对照者。人BAFF重组体(rhBAFF)促进MM细胞的增殖,Si-BCMA可抑制rhBAFF对MM细胞的增殖作用。除了ERK、JNK及p38的表达外,MM细胞中还有活化蛋白p-JNK的表达。Si-BCMA可下调p-JNK的表达,JNK信号通路抑制剂可下调p-JNK及BCMA的表达。结论 MM细胞中,BAFF通过BCMA促进MM细胞增殖。JNK信号通路活化与BCMA表达相互作用,共同促进MM细胞的增殖与存活。     

B淋巴细胞刺激因子及其受体与多发性骨髓瘤

B淋巴细胞刺激因子(BAFF)属于肿瘤坏死因子家族的成员,通过与3个受体(TACI、BCMA、BAFF-R)结合,发挥生物学功能,参与B、T淋巴细胞增殖和功能的调节;BAFF过度表达与受体结合后,可促使B淋巴细胞不断增殖并分泌自身抗体,从而导致一系列自身免疫性疾病的发生;BAFF过度表达,也会使淋巴细胞增殖失控,导致B细胞恶性肿瘤的发生。多发性骨髓瘤是骨髓内浆细胞异常增生的1种B细胞恶性肿瘤,恶性B细胞自分泌产生BAFF,在MM发病过程中起重要作用。     

多发性骨髓瘤患者骨髓中BAFF的含量及B细胞表面BAFF受体表达的研究

本研究旨在分析多发性骨髓瘤(MM)患者骨髓中B细胞活化因子(BAFF)、增殖诱导配体(APRIL)的含量,B细胞、初始B细胞和记忆B细胞表面BAFF受体的表达,以探讨MM骨髓中B细胞的特点。采用流式细胞术检测19例初治MM(初治组)、17例平台期MM(平台期组)、10例对照组骨髓中B细胞(CD19+)、初始B细胞(CD19+IgD+)、记忆B细胞(CD19+CD27+)的表面BAFF受体(BAFF-R、TACI)的表达,ELISA法测定各组骨髓上清中BAFF、APRIL的含量。分析各组结果数据的差异及它们之间的相互关系。结果表明,初治组患者CD19+细胞表面BAFF-R受体表达明显高于平台期组和对照组;初治组患者CD19+IgD+细胞表面BAFF-R受体表达与平台期组无差别,但明显高于对照组;初治组患者CD19+CD27+细胞表面BAFF-R受体表达明显高于平台期组和对照组;各种B细胞表面BAFF-R受体表达在平台期组与对照组之间无差别;TACI受体表达总体水平低,初治组患者与平台期组、对照组比较,CD19+细胞、CD19+IgD+细胞、CD19+CD27+细胞表面TACI受体表达均无明显差异;初治组骨髓上清中BAFF含量明显高于平台期组和对照组,但平台期组与对照组间无明显差异;各组骨髓中APRIL的含量均无明显差异。结论:MM患者骨髓中BAFF含量增高,CD19+细胞、CD19+IgD+细胞和CD19+CD27+细胞表面BAFF受体的表达均增加,这可能有助于促进B细胞的增殖,因而可能与MM发病机制有关。     

B淋巴细胞刺激因子与多发性骨髓瘤关系的研究

B淋巴细胞刺激因子(BLyS)通过和隶属与肿瘤坏死因子受体超家族成员的BCMA,TACI和BLyS-R的结合,在调节外周成熟B淋巴细胞的发育过程中起着重要的作用.由于在多种自身免疫性疾病及肿瘤性疾病中的异常表达,BLyS和其受体也被认为是治疗自身免疫性疾病及肿瘤性疾病的重要治疗靶点.     

儿童急性特发性血小板减少性紫癜患者B淋巴细胞刺激因子和增殖诱导配体表达及意义

目的 探讨B淋巴细胞刺激因子(BLyS)和增殖诱导配体(April)在儿童急性特发性血小板减少性紫癜(ITP)免疫发病机制中的可能作用.方法 急性ITP患儿30例,同龄正常对照儿童30名.采用RT-PCR及实时荧光定量PCR(RQ-PCR)检测BLyS和April及其受体BR3、BCMA、TACI,细胞因子IL-4、IL-5、IL-6、IL-10、IL-15 mRNA表达;流式细胞术检测血小板表面自身抗体PAIgG表达的相对平均荧光强度(MFI).结果 ①急性ITP患儿单核-巨噬细胞(MC)BLyS mRNA[(8.30±2.31)×10-1]和April mRNA[(7.51±1.93)×10-3]表达明显高于同龄正常对照组[分别为(3.95±1.44)×10-1和(3.08±0.82)×10-3](P<0.01);②BLyS和April受体BR3、BCMA和TACI基因转录水平亦明显增高(P<0.01);③急性ITP患儿抗体类型转换相关细胞因子IL-4、IL-5、IL-6、IL-10、IL-15mRNA表达水平明显高于同龄正常对照组(P<0.01);④ITP患儿BLys和April表达调控因子IFN-α、IL-10 mRNA表达水平显著增高(P<0.01);⑤急性ITP患儿PAIgG表达的相对MFI值(67.4±28.1)高于正常对照组(19.5±8.5)(P<0.01),与BLyS和April及其受体(BR3、BCMA和TACI)mRNA的表达呈正相关(r值分别为0.56、0.53、0.62、0.70、0.45,P值均<0.01),激素治疗后明显下降,与BLys和April及其受体表达变化相一致.结论 急性ITP患儿BLyS和April过表达可能是导致ITP免疫功能紊乱的因素之一.     

BLyS and APRIL expression in peripheral blood mononuclear cells of cryptococcal meningitis patients and their clinical significance

ObjectiveTo investigate the levels of APRIL, BlyS and receptors as TACI, BCMA and BAFF-R in peripheral blood mononuclear cells (PBMC) of cryptococcal meningitis (CM) patients and its clinical significance.MethodsPBMC from 30 CM patients and 32 healthy controls were isolated. The mRNA levels of APRIL, BLyS and BLyS receptors were detected by fluorescent quantitation PCR. The effect of PBMC from CM patients on in vitro growth of Cryptococcus neoformans was compared in presence and absence of BLyS.ResultsPBMC of CM patients exhibited significantly lower BLyS, TACI and BCMA mRNA levels but significantly higher BAFF-R mRNA levels than controls. Growth of C. neoformans was significantly slower in presence of BLyS than its absence.ConclusionLevels of BlyS and its receptors correlated with cryptococcal meningitis progression, and provide new clues for monitoring CM conditions and its effective therapy.     

The rGel/BLyS fusion toxin specifically targets malignant B cells expressing the BLyS receptors BAFF-R, TACI, and BCMA

B lymphocyte stimulator (BLyS) is crucial for B-cell survival, and the biological effects of BLyS are mediated by three cell surface receptors designated B cell-activating factor receptor (BAFF-R), transmembrane activator and calcium modulator and cyclophilin ligand interactor (TACI), and B-cell maturation antibody (BCMA). Increased expression of BLyS and its receptors has been identified in numerous B-cell malignancies. We generated a fusion toxin designated rGel/BLyS for receptor-mediated delivery of the recombinant gelonin (rGel) toxin to neoplastic B cells, and we characterized its activity against various B-cell tumor lines. Three mantle cell lymphoma (MCL) cell lines (JeKo-1, Mino, and SP53) and two diffuse large B-cell lymphoma (DLBCL) cell lines (SUDHL-6 and OCI-Ly3) expressing all three distinct BLyS receptors were found to be the most sensitive to the fusion toxin (IC(50) = 2-5 pmol/L and 0.001-5 nmol/L for MCL and DLBCL, respectively). The rGel/BLyS fusion toxin showed specific binding to cells expressing BLyS receptors and rapid internalization of the rGel component into target cells. The cytotoxic effects of rGel/BLyS were inhibited by pretreatment with free BLyS or with soluble BAFF-R, TACI, and BCMA decoy receptors. This suggests that the cytotoxic effects of the fusion toxin are mediated through BLyS receptors. The rGel/BLyS fusion toxin inhibited MCL cell growth through induction of apoptosis associated with caspase-3 activation and poly (ADP-ribose) polymerase cleavage. Our results suggest that BLyS has the potential to serve as an excellent targeting ligand for the specific delivery of cytotoxic molecules to neoplastic B cells expressing the BLyS receptors, and that the rGel/BLyS fusion toxin may be an excellent candidate for the treatment of B-cell malignancies especially MCL and DLBCL.     

Correlation of the expression levels of BLyS and its receptors mRNA in patients with systemic lupus erythematosus

OBJECTIVES: To measure the levels of B-lymphocyte stimulator (BLyS) and its receptors mRNA expression in peripheral blood mononuclear cells (PBMCs) using quantitative real-time polymerase chain reaction (PCR) method and to investigate the relationship between BLyS and its receptors mRNA expression and systemic lupus erythematosus (SLE). DESIGN AND METHODS: Specific primers and TaqMan probe were designed, and real-time PCR was performed. According to the standard curve of plasmids DNA, the levels of BLyS and its receptors mRNA expression in 37 patients with SLE and 30 healthy subjects were determined. The ratio of the expression levels of BLyS mRNA to that of beta2-microglobulin (beta2M) mRNA and the ratio of the expression levels of BLyS receptors mRNA to that of beta2M mRNA were regarded as indicator for the levels of BLyS and its receptors mRNA expression. RESULTS: The expression of BLyS, TACI and BAFF-R mRNA in PBMCs from patients with SLE was significantly elevated compared to healthy controls (P<0.001 for each), and active patients with SLE group had higher mRNA expression than patients with SLE inactive group (P<0.001 for each). The patients with elevated anti-double-stranded DNA (anti-dsDNA) antibody titers had enhanced BLyS, TACI and BAFF-R mRNA expression (P<0.05 for BLyS; and P<0.01 for TACI and BAFF-R). CONCLUSION: The BLyS, TACI and BAFF-R mRNA expression levels were significantly elevated in patients with SLE, which suggests that BLyS, TACI and BAFF-R might be involved in the pathogenesis, and that mRNA expression levels might serve as a biomarker of disease activity.     

多发性骨髓瘤中miR-21与SPRY2基因表达的相关性研究

目的：检测多发性骨髓瘤（MM）患者外周血miR-21的表达水平,研究MM中miR-21与SPRY2基因表达的相关性。方法选择30例MM患者、15例意义未明单克隆丙种球蛋白病（MGUS）患者及20例正常对照（NC）的门诊患者,用实时荧光PCR定量检测miR-21和SPRY2表达水平;Western blot检测miR-21和SPRY2在MM细胞系中表达;在荧光显微镜下观察：U266用脂质体转染荧光标记的 miR-21 mimic/inhibitor后miR-21的表达。结果 MM组血清循环miR-21的表达水平较MGUS组和NC组显著增高,差异有统计学意义（P＜0.01）;在miR-21内源性高表达的MM细胞株中,SPRY2明显低表达;反之,明显高表达;荧光标记的 miR-21 mimic/inhibitor,转染效率90%以上,转染mimics细胞miR-21表达量（98.6±14.2）较未处理的U266细胞miR-21表达量（0.82±0.13）升高了120.2倍（差异有显著的统计学意义,P＜0.001）,转染inhibitor细胞miR-21表达量（0.37±0.06）较未处理细胞降低了61.9%（差异有明显的统计学意义,P＜0.05）。结论 miR-21可能是导致SPRY2在MM中表达下调的一个负性调控因子。miR-21与MM 的发生发展和疾病预后有密切关系,可作为判断MM 患者预后不良指标之一。     

Biochemical markers of bone formation in patients with plasma cell dyscrasias and benign osteoporosis

BACKGROUND: Myeloma-induced bone loss is related to an uncoupling of bone formation and bone resorption. The aim of the present study was to assess the potential clinical value of biochemical markers of bone formation in the work up of patients with plasma cell dyscrasias. METHODS: Serum total alkaline phosphatase, bone-specific alkaline phosphatase (BAP), and osteocalcin (OC) were measured in 43 patients with newly diagnosed multiple myeloma (MM), in 40 patients with monoclonal gammopathy of undetermined significance (MGUS), in 40 patients with untreated benign vertebral osteoporosis (OPO), and in 48 healthy adults. RESULTS: In MM and MGUS patients, serum BAP, but not serum OC, was lower than in healthy controls (P<0.05). Serum OC was higher in patients with OPO than in healthy controls (P<0.05). The strongest associations between markers were found in OPO patients and in healthy adults. MM patients with early-stage disease or without detectable osteolysis had decreased serum BAP values (P<0.05). Serum OC was higher in MM patients with stage III disease (P<0.05) than in healthy controls. MM patients with OPO-like bone involvement had lower BAP values than sex- and age-matched MGUS patients with OPO-like bone involvement and patients with benign OPO (P<0.05). CONCLUSIONS: In patients with plasma cell dyscrasias, serum BAP, rather than serum OC, appears to reflect a suppressed bone formation rate and may be helpful in the differentiation between benign and myeloma-induced OPO. However, the overall clinical use of biochemical markers of bone formation in patients with plasma cell dyscrasia appears limited.     

B细胞成熟抗原在系统性红斑狼疮患者外周血单个核细胞的表达

目的检测B细胞成熟抗原（BCMA）mRNA在系统性红斑狼疮（SLE）患者外周血单个核细胞（PBMC）的表达水平,探讨BCMA在SLE发病中的意义。方法纳入2006年1-11月收治的36例SLE患者,同期17例健康志愿者作为对照组,采用半定量RT-PCR法检测外周血单个核细胞中BCMA mRNA的表达,并与SLE疾病活动指数（SLEDAI）进行相关性分析。结果 SLE患者组BCMA mRNA表达水平（0.598±0.230）均明显高于正常对照组（0.411±0.309）（P〈0.05）。SLE患者BCMA mRNA表达水平与SLEDAI评分无相关性（P=0.590）。结论 SLE患者BCMA mRNA表达水平的增高,可能在SLE的发病机制中具有一定的作用。     

初诊多发性骨髓瘤患者外周血淋巴细胞亚群的检测及意义

目的:通过检测多发性骨髓瘤(multiple myeloma,MM)患者外周血淋巴细胞亚群评价MM患者机体的免疫功能状态并分析其与患者预后的相关性。方法:采用流式细胞术检测32例初诊MM患者和24例健康献血者的外周血T淋巴细胞、B淋巴细胞、NK细胞及CD4~+CD25~+调节性T细胞(CD4~+CD25~+Treg);分别采用溴甲酚绿法、透射免疫比浊法检测患者血清白蛋白(albumin,Alb)、β2-微球蛋白(β2 microglobulin,β2-MG)。结果:与健康献血者比较,MM患者外周血淋巴细胞中CD3~+、CD4~+T淋巴细胞比例无显著改变(P0.05),CD8~+T淋巴细胞、NK细胞比例升高(P0.05),CD4~+/CD8~+T淋巴细胞比值、CD19~+B淋巴细胞比例降低(P0.05),CD4~+CD25~+Treg细胞占CD4~+T淋巴细胞的比例明显升高(P0.01)。CD4~+CD25~+Treg细胞占CD4~+T淋巴细胞的比例与MM的疾病分期呈正相关,与MM患者血清中的β2-MG浓度亦呈正相关(P0.05)。结论:MM患者体内在淋巴细胞亚群的异常表达可能与其肿瘤负荷、病情进展及预后有关。CD4~+CD25~+Treg细胞的表达异常可能是MM免疫逃逸的一个重要机制。