HTLV-1 TaX蛋白与T细胞的细胞周期调变

人1型T细胞白血病病毒(HTLV-1)能引发成人急性T细胞白血病(ATL)以及一种慢性渐进性的中枢神经系统疾病——热性痉挛性下身截瘫/白血病病毒相关脊髓病(TSP/HAM)。成人T细胞白血病(ATL)表现为成熟T淋巴细胞恶性增生。由于HTLV-1 Tax蛋白与T细胞增殖调控有重要关系,本文将主要综述HTLV-1 Tax蛋白如何参与调变T细胞细胞周期从而探讨Tax在T淋巴细胞转化中的作用。     

重组人类T淋巴细胞白血病病毒抗原及双抗原夹心法ELISA抗体诊断试剂盒的研制

为研制灵敏、特异的抗人类T淋巴细胞白血病病毒(HTLV)抗体诊断试剂,将一段HTLV-Ⅰ和HTLV-Ⅱenv区嵌合基因在大肠杆菌中表达,获得的重组抗原具有良好活性.将该抗原作为酶标记抗原建立双抗原夹心法ELISA(dsELISA),对31份HTLV-Ⅰ型血清和19份HTLV-Ⅱ型血清均能100%检出,而在5 065份各种阴性血清中特异度为99.94%.用dsELISA试剂与进口间接法试剂(GeneLabs试剂)平行检测18份HTLV-Ⅰ参比血清、17份HTLV-Ⅱ参比血清和1 024份献血员血清,结果dsELISA试剂正确率为100%,对HTLV-Ⅰ型血清和HTLV-Ⅱ型血清的反应性基本相同,平均s/co值显著高于GeneLabs试剂.而GeneLabs试剂对HTLV-Ⅱ型血清的反应性显著弱于Ⅰ型,并有2份BBI参比血清中的Ⅱ型血清漏检.另外,GeneLabs试剂在献血员血清中出现9例假阳性,特异性显著低于dsELISA试剂.这些结果表明:所研制的dsELISA试剂可用于HTLV-Ⅰ型和Ⅱ型血清的检测,其灵敏度和特异度均优于进口间接法诊断试剂.     

成人T细胞白血病病毒抗体的血清流行病学调查

应用间接免疫荧光试验和明胶凝集试验,对10,013份血清标本进行了人T细胞白血病病毒(HTLV-1)抗体的检测,发现8例阳性。其中3例为日本人,2例是中国台湾人,2例分别是上述日本人和台湾人的妻子,均为中国人,从未离开过大陆;另一例是成人T细胞白血病病人,原为浙江渔民,后当海员,常在日本港口居住。700例各类白血病病人和10例疑似成人T细胞白血病病人的血清,HTLV-1抗体均为阴性。此结果证实,中国大陆正常成年人HTLV-1抗体阴性。少数阳性者均与密切接触日本人有关。HTLV-1病毒是由日本人传入的。     

血友病患者血清中淋巴腺病病毒/人T细胞Ⅲ型病毒抗体检测

对浙江省1982～1984年注射了美国产浓缩Ⅶ因子制剂的18例血友病患者,用酶联免疫吸附法(ELTSA)检测了血清中淋巴腺病病毒/人T细胞Ⅲ型病毒(LAV/HTLV-Ⅲ)抗体,发现4例阳性,并经免疫荧光试验和Western印迹法证实。2例应用了国产浓缩Ⅷ因子者抗体阴性。一例从美国来华旅游死于艾滋病者,LAV/HTLV-Ⅲ抗体阳性。本研究证明,LAV/HTLVⅢ病毒巳通过美国生产的Ⅷ因子制剂传入中国。     

中国HTLV-Ⅰ env基因的克隆及Ⅰ＋Ⅱ型嵌合基因的原核表达与抗原活性检测

为尽快研制出国产HTLV抗体诊断试剂,首先从福建HTLV流行区1名HTLV感染者外周血细胞中克隆出HTLV-Ⅰ的全长膜基因（env）,继而结合文献报道、PSA1软件的新疏水性分析和EPI软件的B细胞表位分析数据,选择了gp46中段开始延伸至gp21N端212个氨基酸（aa185-aa396)的基因,并在3‘端通过（GlySer)2与人工合成的HTLV-Ⅱ型的型特异性表位区基因嵌合,插入原核表达载体pRSET,在E.coli中得到了高效表达目的蛋白产量约占菌体总蛋白的30％。通过Triton-X100洗涤,低湿度尿素逐步变性处理,8mol/L尿素溶解后纯度在75％左右,经电泳洗脱纯化,最终纯度可达95%短期,纯蛋白得率约为40％。经Western blottiong检测,该蛋白对4份HTLV-Ⅰ型和2份HTLV-Ⅱ型均有较强反应,而对4份阴性,血清无反应,从而可能用于研究HTLV抗体诊断试剂盒。     

应用间接免疫荧光试验检测我国正常人和白血病病人血清中嗜T淋巴细胞Ⅲ型病毒抗体

法国巴斯德研究所,自获得性免疫缺损综合症(Aids)先兆症淋巴结病(LAS)分离到一种病毒,命名为淋巴结病相关病毒(LAV);美国Gallo自Aids患者分离出逆转录病毒。后来证明,两者是相同的,但与嗜T淋巴细胞病毒(HTLV)Ⅰ、Ⅱ型不同,称为HTLV-Ⅲ病毒,是Aids的致病因子。 Aids是1981年发现的一种高死亡率疾病。HTLV-Ⅲ病毒侵袭人的OK74或辅助T细胞而     

HLA单倍型与HTLV-1相关疾病

日本鹿儿岛大学医学院病毒学教授园田俊郎于1991年4月5日在病毒学研究所介绍了他们在Ⅰ型嗜人T细胞病毒(HTLV-1)方面的研究进展.     

用杆状病毒载体在昆虫细胞内大量合成Ⅰ型人T细胞白血病病毒功能性P40~x蛋白

人类Ⅰ型T细胞白血病病毒(HTLV-1)是嗜T淋巴细胞病毒,它与一种特殊形式的成人T细胞白血病有关,HTLA-1基因组3′端编码产物-p40~x是一种4万道尔顿的多肽,对HTLV-1长末端重复序列(LTR)内启动子的转录有强的反式激活作用(trans-activation),因而它为病毒复制所必需。 目前已知利用杆状病毒载体表达的外源基因产物有10多种。为了获得足量蛋白以研究p40~x的结构及作     

各地区人类嗜T细胞病毒Ⅰ型的聚类分析

已有相关文献表明人类嗜T细胞病毒Ⅰ型(Human T-lymphotropic virus 1,记为HTLV-Ⅰ)的分布具有区域性,本文旨在提出不同的分析区域性的方法。首先从GenBank中选取来自亚洲、南美洲、非洲的共20条核苷酸序列,用分子生物学软件Vector NTI Suite分析各地区序列样本内部的同源性,然后以各序列的氨基酸含量为对象,定义一个全新的公式进行同源性分析,将该结果与其他研究者采用实验的方法的分析结果比较。结果发现不同的分析方法所得的结论均是一致的。这表明:HTLV-Ⅰ病毒的分布有明显的区域性,文章采用的研究方法对其他流行病学的研究是同样可行的。     

HTLV-1病毒蛋白Tax和HBZ协同促进成人T细胞白血病发生的机制

人类T细胞白血病1型病毒(Human T-cell leukemia virus type 1,HTLV-1)是与人类疾病发生密切相关的逆转录病毒。该病毒的感染可引起成人T细胞白血病(Adult T-cell leukemia,ATL)等多种疾病的发生。Tax和HBZ(HTLV-1 basic zipper protein)是由HTLV-1前病毒编码的两个关键病毒蛋白,它们被认为在HTLV-1病毒的复制、生存和致癌过程中发挥了至关重要的作用。本文就Tax和HBZ的蛋白结构以及它们在调控病毒转录、细胞生长和凋亡、病毒潜伏期,最终协同促进成人T细胞白血病发生过程中发挥的作用作一综述。     

Circulating immune complexes in cerebrospinal fluid of patients with tuberculous meningitis.

Circulating immune complexes (ICs) were isolated from cerebrospinal fluids (CSFs) of patients with tuberculous meningitis (TBM), non-tuberculous neurological diseases by a polyethylene glycol (PEG) precipitation method. Mycobacterium tuberculosis antigen 5 was detected in CICs of 30% patients with TBM, by sandwich ELISA. CIC level decreases during antituberculosis chemotherapy and therefore its detection can provide a method to monitor the therapeutic schedule in patients with TBM.     

Double ligand ELISA technique for the estimation of antibodies to brain tissue antigens in patients with neurological disorders

A double ligand enzyme-linked immunosorbent assay (ELISA) has been developed to detect antibodies against brain tissue antigens in the sera of patients with neurological diseases. The sera were tested on human white matter homogenate. The technique consists of successive incubations with the human serum to be tested, rabbit immunoglobulin G (IgG) to human immunoglobulins (Ig), alkaline phosphate-labeled protein A and alkaline phosphatase substrate. This procedure has the advantage of increased sensitivity compared to the classical ELISA. Application of this procedure to the sera of patients with neurological diseases showed that the unspecific binding is very low and the results are reliable. Moreover the test allows the detection of antibodies to chemically different antigenic structures that can occur in a variety of neurological diseases.     

鞘内注射脐带间充质干细胞治疗神经系统疾病的安全性和疗效研究

目的：探讨腰穿鞘内注射脐带间充质干细胞治疗神经系统疾病的安全性和疗效并且分析腰穿治疗过程中遇到的各种技术 难题。方法：2008 年12 月至2011年5 月88 例伴有神经系统疾病的患者给予鞘内注射干细胞治疗,并且评价其在治疗过程中的 技术难题,采用HAI评分系统对这些患者神经功能的恢复情况进行评估,定期观察患者的临床症状、生物学指标和影像学检查观 察其治疗前后的变化。结果：88 例患者中具有技术难题的患者有20 例,主要表现为在鞘内注射脐带间充质干细胞的过程中需要 行全麻补充、腰穿定位困难等,18 例患者在治疗过程中出现副作用（头痛、低热、腰痛和下肢痛）,但这些副作用在48 小时内经过 系统治疗后完全缓解。随访1 年,50 例患者神经功能得到一定改善,包括15 例脊髓损伤患者、10 例脑瘫患者、10例脑外伤后综合 征患者、5 例脑梗死后综合征患者、5 例脊髓小脑共济失调患者和5 例运动神经元病患者。结论：鞘内注射脐带间充质干细胞治疗 神经系统疾病是安全和有效的,由于随访时间较短,有必要对这种治疗模式进行扩大的、双盲、安慰剂对照研究来进一步推广其 临床应用。     

Free-GABA levels in the cerebrospinal fluid of patients suffering from several neurological diseases Its potential use for the diagnosis of diseases which course with inflammation and tissular necrosis

Summary Free GABA levels were measured in the cerebrospinal fluid (CSF) of 74 neurological patients suffering from cerebral cysticercosis (n = 9), Parkinson's disease (n = 5), multiple sclerosis (n = 6), epilepsy (n = 24), meningeal tuberculosis (n = 6), viral encephalitis (n = 3), cerebrovascular disease (n = 8) and several kinds of dystonia (n = 5). A statistical significant four-fold elevation in free GABA levels was found in patients with cerebral cysticercosis. A non statistical significant two-fold increase in free GABA levels was also encountered in the CSF of patients affected by cerebrovascular disease and viral encephalitis. No changes in CSF free GABA levels were found in patients suffering from any of the other disorders. It is suggested that free GABA levels may be elevated in the CSF of patients suffering from neurological diseases which course with inflammation and tissular necrosis such as cerebral cysticercosis. Much work is needed however to establishd whether CSF free GABA levels can be used as a diagnostic tool in at least some type of these patients.     

5-羟甲基胞嘧啶及其TET氧合酶在神经系统发育和相关疾病中的研究进展

神经系统的正常发育是多种因素相互协调作用的结果,一旦特定因素失衡将引起相关疾病的发生。近年来不断有研究发现,DNA去甲基化过程的一类中间产物5-羟甲基胞嘧啶(5-hydroxymethylcytosine, 5hmC)作为一种新的表观遗传标记,在神经系统中高水平分布,并参与认知、记忆等重要的神经功能。5hmC的形成由氧合酶家族分子(ten-eleven translocation protein, TET)催化,在多种神经系统相关疾病中,5hmC水平和TETs分子的表达都发生改变,提示TET-5hmC表观遗传机制在复杂的神经系统发生发展过程中发挥了重要的调控作用。此外,作为基因表达调控的DNA标记物,5hmC的基因定位与基因表达水平的关系也是重要的研究方向。本文就近年来5hmC和TET家族蛋白分子在神经系统发育和相关疾病方面的重要研究发现进行了综述总结,希望为相关领域研究人员深入开展研究提供重要的思路,并为相关疾病设计治疗策略提供理论支持。     

The Five-Point Test: Reliability,Validity and Normative Data for Children and Adults

The present study provides normative data from a sample of 257 healthy children and 608 adults on a modified version of the Five-Point Test (5PT). The 5PT is a structured and standardized test measuring figural fluency functions. Interrater reliability, test-retest-reliability and construct validity of this measure were analyzed. The sensitivity of the task for cognitive disturbances of patients with neurological diseases was proven by analyzing the test performance in the 5PT of patients with Parkinson''s disease. Finally, normative data stratified by age and corrected for education level is provided. The results of the present study confirm the value of the 5PT in the measurement of figural fluency functions in clinical examination and neuropsychological research.     

ESTIMATION OF GLYCOLIPIDS IN FOUR SELECTED LOBES OF HUMAN BRAIN IN NEUROLOGICAL DISEASES1

Glycolipid (ganglioside, cerebroside and cerebroside sulphate) and cholesterol concentrations for cerebral grey matter from frontal, occipital, temporal and hippocampal lobes of patients with neurological diseases (Alzheimer's disease, senile dementia, cerebrocortical atrophy, schizophrenia and chronic alcoholism) and controls are reported. The results indicate that the concentrations of these lipids are not uniform in the different lobes of both diseased and control brains. The concentrations of the cerebrosides and cerebroside sulphates were generally highest in the occipital lobe and lowest in the frontal lobe; ganglioside N-acetymeuraminic acid (NANA) concentrations on the other hand were lowest in the occipital lobe and highest in the frontal lobe. About one-half of the total NANA was found in the lipid-free residues. There was a general decrease in the concentrations of the glycolipids in the grey matter from the frontal, temporal and hippocampal lobes of brain obtained from patients with neurological diseases (the chronic alcoholic being excluded) below the control values from patients with no known neurological diseases. The cholesterol concentrations in the schizophrenic and alcoholic brains were reduced slightly in all the lobes studied. The general decrease in the glycolipid concentration in the diseased brain may indicate the extent of cortical degeneration.     

High Titers of Autoantibodies to Glutamate Decarboxylase in Type 1 Diabetes Patients: Epitope Analysis and Inhibition of Enzyme Activity

ObjectiveAutoantibodies to glutamate decarboxylase (GAD65Ab) are found in patients with autoimmune neurological disorders or type 1 diabetes. The correct diagnosis of GAD65Ab-associated neurological disorders is often delayed by the variability of symptoms and a lack of diagnostic markers. We hypothesized that the frequency of neurological disorders with high GAD65Ab titers is significantly higher than currently recognized.MethodsWe analyzed GAD65Ab titer, GAD65 enzyme activity inhibition, and GAD65Ab epitope pattern in a cohort of type 1 diabetes patients (n = 100) and correlated our findings with neurological symptoms and diseases.ResultsOverall, 43% (43/100) of patients had detectable GAD65Ab titers (median = 400 U/mL, range: 142250,000 U/mL). The GAD65Ab titers in 10 type 1 diabetes patients exceeded the 90^th percentile of the cohort (2,000250,000 U/mL). Sera of these 10 patients were analyzed for their GAD65Ab epitope specificity and their ability to inhibit GAD65 enzyme activity in vitro. GAD65Ab of 5 patients inhibited the enzyme activity significantly (by 34-55%). Three patients complained of muscle stiffness and pain, which was documented in 2 of these patients.ConclusionsBased on our findings, we suggest that neurological disorders with high GAD65Ab titers are more frequent in type 1 diabetes patients than currently recognized. (Endocr Pract. 2013;19:663-668)     

Detection of IgG antibody to measles virus by radioimmunoassay technique

A highly sensitive procedure of solid-phase radioimmunoassay (RIA) was developed for the detection of measles IgG antibody. HeLa cells persistently infected with measles virus were used as a solid-phase antigen. This technique was applied to the detection of measles IgG antibody in patients with subacute sclerosing panencephalitis (SSPE) and multiple sclerosis. Normal subjects having experienced natural measles or measles vaccination and patients with various neurological diseases of non-virus nature were also examined as control groups. Measles antibody was detected at high titers in both the sera and cerebrospinal fluid of SSPE patients. Moreover, RIA/HI ratios of SSPE patients were significantly higher than those of normal subjects, suggesting the presence in the formers of antibodies to nucleocapsids at high titers as well as to viral envelopes. On the other hand, no significant difference was found in both RIA and HI titers between the sera of multiple sclerosis and those of various neurological diseases.     

Anti-galactocerebroside antibodiesin human cerebrospinal fluids determined by enzyme-linked immunosorbent assay (ELISA)

The standard ELISA technique was improved for the detection of antigalactocerebroside antibody in biological fluid. Mouse monoclonal antigalactocerebroside antibody was used to demonstrate specificity and sensitivity of the technique. After optimization of the assay, the usefulness of this measurement for the evaluation of patients with multiple sclerosis was assessed. The presence of antigalactocerebroside antibodies in the cerebrospinal fluid of 20 patients with multiple sclerosis, 10 with other neurological diseases and 10 normal individuals was determined. All the CSF samples from normal individuals were negative. In patients with multiple sclerosis 14 of the 20 samples had elevated levels of antigalactocerebroside antibody, whereas with other neurological diseases 5 out of 10 were positive. Antigalactocerebroside levels were lower in samples from patients during an acute relapse than in those from more chronic cases. These results indicate that the presence of anti-galactocerebroside antibody in cerebrospinal fluid is not specific to MS but may reflect previous damage to myelin.Abbreviations and trivial names used ELISA Enzyme-Linked Immunosorbent Assay - CSF cerebrospinal fluid; galacto- or glucocerebroside, ceramide-1-0-beta-galactoside or-glucoside