T cell clones expressing the natural killer cell-related p58 receptor molecule display heterogeneity in phenotypic properties and p58 function

A new anti-p58 monoclonal antibody (mAb), termed CH-L, has been used to characterize a minor subset of T lymphocytes co-expressing p58 and CD3 molecules. In two-color immunofluorescence analysis, CH-L⁺CD3⁺ cells represented 0.5 to 6% of the peripheral blood lymphocytes (in 20 healthy donors). Clonal analysis showed that most CD3⁺CH-L⁺ T cell clones expressed the CD8⁺4^? T cell receptor (TcR) α/β⁺ phenotype, while only a few were CD8^?4⁺ TcR α/β⁺, CD8^?4^? TcR α/β⁺ or CD8^?4^? TcR γ/δ⁺. Western blot analysis indicated that the CH-L mAb identifies the same 56-58-kDa diffuse band in both T and natural killer cell (NK) clones. A minority of T cell clones also expressed other NK-related markers such as CD16, CD56 and CD94 and two clones also reacted with the anti-p58 mAb EB6. Interestingly, most clones displayed cytolytic activity in an anti-CD3 mAb-triggered redirected killing assay against the Fcδ receptor⁺ P815 target cells and NK-like activity against K562 and Raji cells. In contrast, the IGROV-1 ovarian carcinoma cell line was resistant to cytolysis by all of these clones. Since p58 molecules have previously been shown to exert regulatory functions on NK-mediated lysis, we investigated whether anti-p58 mAb could also influence cytotoxicity mediated by CD3⁺p58⁺ T lymphocytes. Lysis of P815 target cells, triggered by anti-CD3 mAb, could be inhibited by anti-p58 mAb in 8 out of 12 cytolytic clones tested, while 4 clones were not inhibited. In addition, anti-p58 mAb enhanced the cytolytic activity of 3 clones against IGROV-1 and of 4 other clones against Raji target cells. Taken together, these data indicate that p58⁺ T cells express heterogeneous phenotypes and different forms of TcR and, in most instances, display cytolytic functions. Perhaps more importantly, the p58 molecule appears to modulate the cytolytic activity triggered via the CD3/TcR complex.     

淋巴细胞功能相关抗原-3(CD58)在儿童期急性B淋巴细胞白血病近期疗效判断中的应用

本研究评估CD58在急性B淋巴细胞白血病(B-ALL)近期疗效判断中的价值。应用四色流式细胞分析技术观察135例儿童期B-ALL患者CD58分子的表达特点;建立用CD58/CD10/CD34/CD19抗体组合检测B-ALLMRD的方案;结合CD58的表达情况和MRD监测结果分析CD58在B-ALL中的预后价值。结果表明135例B-ALL的平均CD58MFI为113.08±63.33,15例正常骨髓的CD19+CD10+细胞的平均CD58MFI为14.68±5.26,两者差异显著(P<0.01);51.9%(70/135)B-ALL患者的CD58分子强表达,可以用CD58为指标进行MRD检测;CD58/CD10/CD34/CD19抗体组合的有效频率仅次于TdT/CD10/CD34/CD19,为51.9%;CD58高表达组的MRD阳性率显著低于CD58低表达组(P<0.05)。结论CD58可以作为B-ALLMRD检测的指标,此结果丰富了MRD检测的组合;CD58的高表达可以作为B-ALL预后较好的指标。     

人原发性肺癌组织中TSLC1基因甲基化检测及意义

目的探讨TSLC1基因启动子区甲基化状态与人原发性肺癌发生发展之间的关系方法采用甲基化特异性聚合酶链技术(MSP),对53例人原发性肺癌组织(肺癌组)、24例肺良性病变组织(良性病变组)、15例癌旁组织(癌旁组)及11例支气管鳞状化生组织进行TSLC1基因5’-CpG岛甲基化状态检测。结果肺癌组、癌旁组和肺良性病变组的TSLC1基因甲基化率分别为35.8%(19/53)、13.3%(2/15)、0.0%(0/24),三组相比差异有统计学意义(P<0.05);支气管鳞状化生组织的甲基化阳性率为18.2%(2/11)。TSLC1基因甲基化率肺癌吸烟者与非吸烟者差异无统计学意义(P>0.05)。结论 TSLC1基因启动子区甲基化与肺癌的发生关系密切,可能出现在肺癌发生的早期阶段。     

坐下-站立功能测试训练系统数据采集与无线通讯模块的开发

目的:研发一套基于MSP430单片机与蓝牙无线通信的用于人体坐下-站立功能的测试系统。方法:系统采用上下位机结构,通过压力传感器采集坐下-站立过程中的足底和臀部的压力参数,通过蓝牙无线传输将数据传送至上位机。仪器研制成功后,对20位志愿者进行了坐下-站立功能的测试。结果:设计出该系统。利用上位机开发的测试训练软件,实现压力变化的实时显示及评定结果的数值显示。20位志愿者坐下-站立功能测量所得各参数的组内相关系数(ICC)均大于0.8,表明该系统具有很好的可靠性。结论:本研究所设计的坐下-站立功能测试系统可定性定量地测试人体坐下-站立的平衡功能。     

白血病细胞系中抑癌基因PTEN启动子区甲基化状态检测及其去甲基化研究

目的 探讨抑癌基因PTEN表达沉默的机制及诱导PTEN表达对白血病细胞的作用.方法 应用甲基化特异性聚合酶链反应(Methylation specific PCR,MSP)检测白血病细胞系HL-60、Nalm-6、Raji、KG-1a、U937、NB4、K562中PTEN基因启动子区域甲基化状态;用甲基化转移酶抑制剂5-氮-2'-脱氧胞苷(5-Aza-CdR)处理白血病细胞,MSP检测甲基化状态的改变、RT-PCR检测PTEN mRNA水平的改变、瑞特染色观察细胞形态学改变、膜联蛋白V/碘化丙锭(Annexin V/PI)标记染色检测细胞凋亡.结果 检测的白血病细胞系中HL-60、Nalm-6、Raji、KG-1a、U937细胞PTEN基因显示超甲基化状态,而NB4和K562细胞显示低甲基化状态;5-Aza-CdR处理HL-60和Nalm-6细胞后,PTEN基因甲基化降低、mRNA表达水平则逐渐增高,并呈剂量依赖性,细胞出现凋亡现象.结论PTEN基因启动子区异常甲基化可能导致该基因转录表达失活或沉默,甲基化抑制剂可以诱导PTEN表达,并引起白血病细胞凋亡.     

猪带绦虫TSO45W-4BX与猪CD58分子在大肠埃希菌中的联合表达

目的 以猪CD58为分子佐剂，将CD58基因与猪带绦虫疫苗候选抗原基因TSO45W-4BX联合表达，寻找新型抗猪囊尾蚴疫苗。 方法 分别以重组质粒pGEM-4B和pGEM-CD58为模板，PCR扩增猪带绦虫TSO45W-4BX基因和猪CD58基因，将TSO45W-4BX与酶切处理的pGEX-4T-1定向连接，转化大肠埃希菌JM109，重组质粒经鉴定正确后，在其下游酶切插入CD58基因，PCR扩增和测序证明阅读框正确后，用异丙基-β-D-硫代半乳糖苷（IPTG）诱导表达，用十二烷基磺酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE））和蛋白质印迹法（Western blotting）分析表达产物的免疫活性。 结果 pGEX-4BX和pGEX-4BX/CD58分别表达Mr 41 000和Mr 69 000的融合蛋白，pGEX-4BX表达产物主要以可溶性形式存在，而pGEX-4BX/CD58却以包涵体形式存在，但两者都能被囊尾蚴病患者血清识别。 结论 TSO45W-4BX与CD58基因联合表达，TSO45W-4BX仍具有免疫活性。     

Association of JC virus T-antigen expression with the methylator phenotype in sporadic colorectal cancers

BACKGROUND & AIMS: JC virus (JCV) is a polyomavirus that ubiquitously infects humans and has been implicated in various human cancers. JCV encodes a "transforming" gene, T-antigen (T-Ag), which is believed to mediate the oncogenic potential of the virus. We have previously shown that JCV DNA sequences are usually present in human colorectal cancers (CRCs), and we have provided in vitro evidence that JCV can induce chromosomal instability (CIN) in CRC cells. This study tests the hypothesis that JCV T-Ag expression correlates with one or more forms of genomic or epigenetic instability in sporadic CRCs. METHODS: We characterized 100 sporadic CRCs for microsatellite instability (MSI) and CIN. PCR amplifications were performed for T-Ag sequences, and immunohistochemical (IHC) staining was performed to detect T-Ag expression. De novo methylation of the promoter regions of nine putative tumor suppressor genes thought to play a role in colorectal carcinogenesis was studied by methylation-specific PCR. RESULTS: JCV T-Ag DNA sequences were found in 77% of the CRCs and 56% of these cancers (or 43% of the total) expressed T-Ag by IHC. Significant associations were observed between T-Ag expression and CIN in CRCs (P = .017) and between T-Ag expression and promoter methylation of multiple genes (P = .01). CONCLUSIONS: The association between T-Ag expression and promoter methylation in CRC suggests that this viral oncogene may induce methylator phenotype and that JCV may be involved in CRC through multiple mechanisms of genetic and epigenetic instability.     

西安地区耐亚胺培南鲍曼不动杆菌的碳青霉烯酶及同源性研究

目的 调查西安地区耐亚胺堵南鲍曼不动杆菌的耐药性,研究其同源性及分子耐药机制.方法 收集西安地区6所三级甲等医院1年间临床分离的非重复鲍曼不动杆菌株146株,采用K-B法进行药敏试验,E试验检测金属酶(MBL),PCR扩增OXA-23,-24,-58,-51 like型及IMP-1型和VIM-2型碳青霉烯酶基因,其阳性产物经测序分析,应用肠杆菌科基因组内重复序列(ERIC)-PCR对菌株进行DNA分型和同源性分析.结果 筛选出对亚胺培南耐药鲍曼不动杆菌(IRAB)非重复株1 5株,其中1株经E试验检测产金属酶,但扩增blaTMP-1,blaVIM-2均阴性,另外14株扩增blaOXA-66均阳性,11株blaUXA-23阳性,1株blaXA 58阳性;ERIC-PCR将15株IRAB分为A型和B型,其中部分菌株的亲缘关系很近,同源性达90%以上.结论 产OXA型碳青霉烯酶是西安地区该菌对亚胺培南耐药的主要原因.其中OXA-23型普遍存在,OXA-58型和OXA-66型基因在国内尚属新型碳青霉烯酶基因型;15株IRAB为2种克隆型,可能存在局部暴发流行.     

Epstein-barr virus-related gastric adenocarcinoma: an early secondary cancer post hemopoietic stem cell transplantation

BACKGROUND & AIMS: Epstein-Barr virus (EBV) infection has been associated with some cases of gastric cancer. METHODS: We studied a case of early onset gastric adenocarcinoma after nonmyeloablative hematopoietic stem cell transplantation for myeloma in a 56-year-old man. RESULTS: The development of gastric adenocarcinoma was preceded by severe graft-versus-host disease (GVHD) necessitating strong immunosuppression, which resulted in an intense reactivation of EBV infection. Three sequential gastric biopsy examinations performed at 100, 130, and 150 days after hematopoietic stem cell transplantation showed gastritis, dysplasia, and adenocarcinoma, respectively. There was no evidence of Helicobacter pylori infection. Quantitative polymerase chain reaction for circulating EBV showed a surge of EBV DNA peaking at the time of gastritis, followed by a gradual decrease afterward with adequate control of GVHD and tailing of immunosuppression. In situ hybridization for EBV-encoded early small RNA showed absence of EBV in the gastritis specimen, but the presence of EBV in the dysplastic and carcinoma specimens. Aberrant promoter methylation of E-cadherin was observed only in the carcinoma specimens, showing that infection with EBV preceded E-cadherin methylation. CONCLUSIONS: Mucosal damage caused by GVHD, immunosuppression, and EBV reactivation combined to lead to EBV infection of the gastric cells and initiation of carcinogenesis, suggesting this case to be a genuine EBV-related opportunistic malignancy post-transplantation. An interesting proposition is that this case also might reflect a compacted timeline of events in EBV-related gastric cancers developing in immunocompetent patients.     

恶性疟原虫裂殖子表面蛋白-2DNA疫苗诱导小鼠T淋巴细胞的增殖

目的　为了探讨恶性疟原虫FCC - 1/HN株裂殖子表面蛋白 - 2 (MSP - 2 )DNA疫苗在小鼠体内诱导的免疫应答的特性及抗感染的保护性免疫机制。方法　将重组真核表达质粒pBK/MSP - 2经骨骼肌注射BALB/c小鼠 ,小鼠经DNA疫苗免疫 8w后 ,用流式细胞仪分析脾脏T淋巴细胞的分化 ,并体外培养脾脏细胞 ,用夹心ELISA法测定IFN -γ和IL - 2的产生。结果　与对照组相比 ,疫苗组CD+ 4CD8+ T淋巴细胞有显著性的增高 ,体外培养的脾脏细胞IFN -γ有一个高浓度的分泌。结论　恶性疟原虫FCC - 1/HNMSP - 2DNA疫苗诱导了一个TH1的免疫应答类型