恒河猴肝移植模型急性排斥反应中白细胞介素6的表达

背景：白细胞介素6是免疫炎症反应中一种重要的细胞因子,它与移植排斥反应具有密切的关系,在移植排斥反应诊断和抗排斥疗效评价中发挥着重要作用。 目的：检测白细胞介素6在恒河猴肝移植后急性排斥反应中的表达水平,以评价其作为肝移植后急性排斥反应早期诊断指标的价值。 方法：以16只恒河猴为研究对象,将其随机分为实验组(围手术期不给予免疫抑制治疗)和对照组(围手术期给予免疫抑制治疗),然后建立恒河猴同种异体原位肝移植模型;分别在移植后6,12,24,72 h时间点收集血清及肝脏组织,通过检测肝功指标和移植肝组织苏木精-伊红染色Banff评分反映其移植排斥反应情况,并应用酶联免疫吸附法、免疫组织化学技术分别检测白细胞介素6的表达水平。 结果与结论：肝移植后12,24,72 h实验组均有急性排斥反应发生,其中实验组丙氨酸氨基转移酶、天冬氨酸氨基转移酶、总胆红素在24 h和72 h表达水平明显高于对照组(P < 0.05);实验组72 h组织学表现重于对照组,Banff评分高于对照组(P < 0.05);实验组中血清、肝组织的白细胞介素6水平在肝移植后12,24,72 h明显高于对照组(P < 0.05),并以72 h最为明显。结果表明白细胞介素6可能参与肝移植后排斥反应的发生,其表达可能在恒河猴原位肝移植后急性排斥反应的早期诊断中有一定意义。 中国组织工程研究杂志出版内容重点：肾移植;肝移植;移植;心脏移植;组织移植;皮肤移植;皮瓣移植;血管移植;器官移植;组织工程全文链接：     

恒河猴肝移植急性排斥反应中核因子κB p65的表达

背景：核因子κB作为一种重要的核内转录因子,是多种信号转导途径的汇聚点,参与机体免疫细胞的增殖、分化及细胞凋亡等多种反应物质基因的表达调控,在体液和细胞免疫中发挥重要作用。 目的：探讨恒河猴移植肝组织内核因子κB P65蛋白表达与急性排斥反应的关系。 方法：将恒河猴随机分为2组：急性排斥反应组肝移植后不给予抗排斥处理,对照组肝移植过程中及移植后均给予抗排斥处理。分别在移植后6,12,24和72 h 4个时间点收集血标本,全自动生化分析仪测定丙氨酸氨基转移酶、总胆红素,取移植肝脏组织行苏木精-伊红染色观察组织形态结构和排斥反应,根据Banff评分系统判断排斥反应程度,采用Western blot法检测肝脏组织中核因子κB p65的表达。 结果与结论：肝移植急性排斥反应发生时肝功能变化滞后于肝组织病理学检查。当急性排斥反应发生时,移植肝组织中核因子κB p65表达上调,急性排斥反应程度也随之加剧。并且在急性排斥反应早期,肝功能、病理学仅有轻微改变时,核因子κB p65表达显著升高。因此移植肝组织中核因子κB p65表达水平的检测对移植后急性排斥反应的早期诊断有重要意义,同时核因子κB可能成为控制移植急性排斥反应的新靶点。     

热休克蛋白70与肝癌

热休克蛋白70(HSP70)广泛存在于细胞中,在应激刺激后过表达,可以参与蛋白质的合成及受损蛋白的修复。HSP70在肝癌治疗中的作用日益受到重视:利用其特异性和免疫原性,靶向治疗肝癌。并且可以利用其分子伴侣的特点,作为肝癌疫苗,诱发机体抗肝癌的免疫反应。HSP70在肝癌治疗中已初显成效,但也存在很多问题,需要更加深入的研究。总之,HSP70在肝癌治疗中有很大的应用前景。     

大脑局部急性缺血后热休克蛋白70的免疫组织化学研究

热休克蛋白７０（ＨＳＰ７０）是一种极敏感和可靠的脑缺血的标志。在正常脑内几乎检测不到ＨＳＰ７０ｍＲＮＡ或ＨＳＰ７０蛋白，随着脑缺血时程的增长，ＨＳＰ７０蛋白的表达与细胞受缺血损伤的程度相关，但随着缺血程度加重，ＨＳＰ７０基因转录和／或翻译被阻断。一过...     

热休克诱导小鼠肝细胞表达热休克蛋白70的研究

目的：研究热休克恢复期小鼠肝细胞热休克蛋白（ＨＳＰ７０）的表达。方法：小鼠分别为４０、４２、４４、４６℃热休克处理３０ｍｉｎ,恢复８ｈ;４６℃热休克处理３０ｍｉｎ,然后分别于热休克后２－７２ｈ取肝,以免疫组化方法观察ＨＳＰ７０的表达,并用体视学方法对阳性肝细胞体密度进行定量分析。结果：４２、４４、４６℃均能诱导肝细胞表达ＨＳＰ７０,以４６℃组小鼠阳性肝细胞密度最大;４６℃热休克后８－１２ｈ为ＨＳＰ     

妊娠早期小鼠子宫内膜热休克蛋白70的免疫组化研究

目的 :研究妊娠早期小鼠子宫内膜热休克蛋白 70的表达。方法 :采用免疫组织化学方法。结果 :热休克蛋白 70主要存在于子宫内膜固有层的基质细胞及蜕膜细胞 ,内膜上皮、腺上皮中未见表达。与未孕小鼠相比 ,孕鼠热休克蛋白 70免疫反应阳性细胞显著增多 (P <0 .0 1 )且随妊娠日龄的增加而增加 (P <0 .0 1 )。结论 :热休克蛋白70可能参与了子宫内膜蜕膜反应中基质细胞的增殖 ,与蜕膜反应密切相关     

细胞外热休克蛋白70及其生物学功能

热休克蛋白(HSP)一直被描述为一种细胞内蛋白质在胞内发挥一系列生物学作用,参与细胞内蛋白质的折叠、装配、降解和修复过程。近年来,有研究发现HSP70能被主动释放到胞外,成为细胞外HSP70(extracellular HSP70,eHSP70),但关于其功能尚不清楚。循环HSP70水平与多种疾病进程密切相关。有研究发现暴露于物理和心理刺激源作用下,HSP70可能通过脂筏或Exosome介导的分泌途径释放到细胞外,作为一种生物活性因子影响多种疾病的进程。     

中国肝移植后排斥反应研究的发展趋势

背景：终末期肝病患者可以通过移植全部肝脏或部分肝脏来挽救生命,随着新型免疫抑制药物的出现,肝移植后排斥反应的发生率大幅度降低,使患者的生存率得到明显提高。 目的：对肝移植后排斥反应研究的文献资料趋势进行多层次探讨分析。 方法：以电子检索方式对CNKI数据库学术期刊和博士学位论文数据库2002-01/2011-12收录有关肝移植后排斥反应研究的文献进行分析,采用检索词为“肝移植;排斥反应”,应用数据库的分析功能和Excel软件图表的功能分析数据特征。 结果与结论：在CNKI数据库学术期刊2002/2011收录的文献中,共检索到777篇与肝移植后排斥反应研究相关的文献,从文献数量上看,2006年的文献数量处于顶峰为110篇,2007年《人体器官移植条例》颁布以前处于上升趋势,到2007年开始略有下降。文献的基金资助项目数量比较多,39个基金资助项目文献共有210篇,省级基金资助项目数量为28个,其中广东省的基金项目最多为4个。《中华器官移植杂志》是中国肝移植后排斥反应研究的权威期刊。肝移植后排斥反应的研究主要以大鼠的动物实验为主,移植后急性排斥反应的发生率比较高,强效的新型免疫抑制剂以他克莫司的研究较多。在博士学位论文数据库中检索到90篇相关文献,文献数量、学科类别、研究机构、关键词和基金资助项目结果与CNKI数据库学术期刊文献结果基本相似。     

人热休克蛋白70基因的原核表达

目的 表达人热休克蛋白70（HSP70）并进行鉴定。方法 用PCR方法扩增人HSP70基因片段，经T-A克隆法克隆到载体pUCm-T中，并进行DNA测序，将人HSP70基因片段从载体pUCm-T中酶切后，构建重组表达载体pGEX-4T-1-HSP70，并转化大肠杆菌JM109，用IPTG诱导，收集细菌，菌体裂解后进行SDS-PAGE及Western blot检测。结果 人HSP70基因的PCR产物约为1.9kb。序列测定结果证实，所获目的序列与文献[3]报道的相一致，经EcoRⅠ和XhoⅠ酶切鉴定证实。人HSP70基因已成功地克隆到表达载体pGEX-4T-1中，构建的表达载体pGEX-4T-1-HSP70，能很好地在大肠杆菌中表达相对分子质量（Mr)为96000并具有抗原特性的融合蛋白，结论 成功地克隆并表达了HSP70基因，为研究HSP70的结构。功能与临床应用提供了必要条件。     

恒河猴移植肝组织内细胞间黏附分子1与急性排斥反应的关系

背景：在急性排斥反应发生时会伴有移植组织中细胞间黏附分子1表达水平的增高。 目的：探讨恒河猴移植肝组织内细胞间黏附分子1与急性排斥反应的关系。 方法：建立恒河猴同种异体肝移植模型,随机分为实验组(移植后不给予抗排斥处理)和对照组(移植中及移植后均给予抗排斥处理)。 结果与结论：实验组移植后24,72 h血清丙氨酸氨基转移酶及血清总胆红素水平明显高于对照组(P < 0.05或< 0.001)。移植后12 h实验组移植肝即表现为轻度急性排斥反应,说明急性排斥反应时肝功能变化滞后于肝组织病理学检查,移植后24,72 h表现为中重度急性排斥反应。移植后6 h实验组肝组织中血管内皮细胞、肝细胞膜、胆管上皮细胞上细胞间黏附分子1的表达呈持续增强,并显著高于对照组(P < 0.05),随细胞间黏附分子1的表达增强,排斥反应加剧,说明在急性排斥反应早期,肝功能、病理学仅有轻微改变时,细胞间黏附分子1水平即显著升高。提示移植肝组织中细胞间黏附分子1表达的检测对肝移植后急性排斥反应的早期诊断具重要意义。     

Distribution of 70kDa heat shock protein in rabbit brains after heat stress and heat stroke

OBJECTIVE: Evaluation of the relationship between the induction of 70kDa heat shock protein in rabbit brains and heat stress. METHODS: HSP70 was detected using monoclonal antibody by ABC method in rabbit hypothalamus, hippocampus and cerberal cortex. RESULTS: Intense HSP70 staining was displayed in rabbit brains of the heat stroke group (rectal temperature 43 degrees C to death). Positive cells were distributed mainly in the CA1, CA2 regions of the hippocampus; granular cell layer I and pyramidal layer (II) of the cerebral cortex; and the periventricular area of hypothalamus. HSP70-psoitive substances were localized in the cytoplasm and neuronal processes, a few neurons exhibited dark staining nucle. Hosever, the rabbit brains of the general heat stress group (rectal temperature 42.0 degrees C, 30 minutes) had much weaker staining. CONCLUSION: Hyperthermia causes neuronal expression of HSP70, particularly under strong heat stress, and may be sustained till death.     

CD28 gene polymorphisms and acute cellular rejection after liver transplantation

The co-stimulatory molecule CD28 plays an important role in T-cell-mediated immune response like acute cellular liver transplant rejection. The aim of the retrospective case- control study was to examine whether the single nucleotide polymorphisms (SNPs) rs3116487, rs3116494, and rs3116496 of the CD28 gene are associated with acute cellular liver transplant rejection. The mentioned SNPs were genotyped in 147 liver transplant recipients without acute cellular rejection and 144 liver transplant recipients with acute cellular rejection by real-time endpoint genotyping. The genotype and allele frequencies of the SNPs did not show any significant differences between both groups. Haplotype analyzes of the SNPs also showed no association. Our data suggest that the analyzed SNPs are not major contributors to the susceptibility of acute cellular liver transplant rejection.     

Immunocytochemical detection of the 70-kd heat shock protein in alcoholic liver disease.

Exposure of cells to physical (eg, heat) or chemical (eg, alcohol) stress results in increased synthesis of a set of highly conserved polypeptides termed heat shock proteins (HSPs), among which the 70-kd protein (HSP 70) is one of the most consistently inducible and highly conserved. This HSP has adenosine triphosphate-binding properties and is known to associate strongly with cytoskeletal structures that are usually disrupted on injury by heat or alcohol. Some HSPs apparently function as accessories to a nonlysosomal, adenosine triphosphate-dependent proteolytic system that binds and digests away stress-generated abnormal or denatured proteins after their conjugation with ubiquitin, a small HSP. Ubiquitin has been demonstrated immunocytochemically in Mallory bodies, which represent mainly degenerated intermediate filaments accumulated in hepatocytes of alcoholic-diseased liver. We immunostained histologic sections from patients with alcoholic liver disease using a polyclonal antibody raised against HSP 70. Strong diffuse cytoplasmic immunoreactivity was observed in many hepatocytes, including cells without Mallory bodies or fatty degeneration. Positive immunoreactivity for HSP 70 points to a possible involvement of this HSP in the pathogenesis of alcoholic liver disease. It also suggests that immunocytochemical detection of HSP 70 may serve as a more sensitive indicator of hepatocellular injury.     

Immunohistochemical expression of heat shock protein 70 in vitiligo

Heat shock proteins (HSPs) are proteins that are expressed under variety of stresses including pathologic conditions. How stresses affect vitiligo is not fully understood and little is known about the role of HSPs generally and Hsp70 specifically in vitiligo. The current study investigated the expression of Hsp70 in vitiliginous (32) and normal skin (10) by immunohistochemistry together with correlating this expression with the clinicopathologic parameters in the studied vitiligo group. Hsp70 was expressed in the cytoplasm of epidermis in all normal skin compared with its localization to the cytoplasm in 35.5% and to the nuclei in 64.5% of epidermis in vitiligo lesions. Intense (P < .001) and diffuse (P < .001) expression of Hsp70 was in favor of vitiligo skin compared with normal skin. Nuclear form of Hsp70 tended to be expressed in progressive forms of the disease. The percentage of Hsp70 expression tended to be decreased with the duration of the disease. From the present study, up-regulation of HSP 70, in the form of its intense and diffuse expression, may be blamed in pathogenesis of vitiligo. Nuclear localization of HSP 70 may be more important than its presence or absence, beside it may be related to progression of the disease.     

Facets of heat shock protein 70 show immunotherapeutic potential

Amongst the families of intracellular molecules that chaperone and assist with the trafficking of other proteins, notably during conditions of cellular stress, heat shock protein (hsp) 70 is one of the most studied. Although its name suggests that expression is exclusively induced during cellular hyperthermia, members of the hsp70 family of proteins can be constitutively expressed and/or induced by a range of other cellular insults. The ubiquitous presence of hsp70 in eukaryotic and prokaryotic cells, combined with its high degree of sequence homology and intrinsic immunogenicity, have prompted the suggestion that inappropriate immune reactivity to hsp70 might lead to pro-inflammatory responses and the development of autoimmune disease. Indeed, hsp70 has been shown to be a potent activator of innate immunity and aberrant expression of hsp70 in certain organs promotes immunopathology. However, studies also suggest that hsp70 might have immunotherapeutic potential, as hsp70 purified from malignant and virally infected cells can transfer and deliver antigenic peptides to antigen-presenting cells to elicit peptide-specific immunity and, in contrast to its reported pro-inflammatory effects, the administration of recombinant hsp70 can attenuate experimental autoimmune disease. This review focuses on the immunoregulatory capacity of hsp70 and its potential therapeutic value.     

颗粒酶B和穿孔素免疫组织化学检测诊断肝移植急性排斥反应的敏感性与特异性

目的　探讨颗粒酶B和穿孔素两种免疫活化分子在肝移植急性排斥诊断中的作用,及其与Banff急性排斥反应组织学诊断标准之间的对应关系。方法　在常规组织学诊断基础上,将41份肝穿刺标本用颗粒酶B与穿孔素单克隆抗体进行免疫组织化学EnVision二步法标记,IPP图像分析软件计算阳性细胞数/mm2 作为免疫活化细胞指数(AI),以组织学诊断作为评判有无急性排斥反应的标准。结果　在41份肝穿刺标本中,组织学诊断为急性排斥反应21份,缺乏急性排斥反应组织学改变20份。急性排斥反应组颗粒酶B与穿孔素AI值显著高于无排斥反应组(P<0. 001),中重度排斥反应组AI值显著高于轻度及其非确定性排斥反应组(P<0. 001)。与组织学诊断比较,颗粒酶B的敏感性、特异性、阳性预测值、阴性预测值及诊断一致率分别达到90. 0%、95. 2%、94. 7%、90. 9%以及92. 7%;穿孔素的各指标也分别达到80. 0%以上。结论　颗粒酶B与穿孔素是急性排斥反应免疫效应细胞活化标志,在临床肝移植急性排斥反应时表达明显升高,作为组织学诊断急性排斥反应的辅助指标具有相当高的敏感性及特异性,可用于肝移植后肝穿刺标本的鉴别诊断。     

Identification of heat shock protein 70 in the rabies virion.

We investigated a 73-kDa polypeptide (p73), a minor component of the rabies virion (HEP-Flury and ERA strains), accounting for as much as 1% of total virion proteins. Two-dimensional gel electrophoresis and immunoblotting with the antiserum against the heat shock protein 70 (hsp70) demonstrated that p73 was identical to a constitutive type of cellular hsp70. The antiserum also detected p73/hsp70 in the purified virions of other negative-stranded RNA viruses, such as vesicular stomatitis virus (New Jersey serotype), Newcastle disease virus (Miyadera strain), and influenza A virus (PR8 strain), among which, however, the contents were variable.