γδT细胞识别人多发性骨髓瘤细胞XG-7膜热休克蛋白-70

目的：探讨人多发性骨髓瘤细胞（ＭＭ）ＸＧ－７选择性激发扩增人外周血γδＴ细胞机制。方法：运用蛋白质分离纯化手段获取ＭＭ细胞ＸＧ－７膜蛋白组分,检查每一组分对γδＴ细胞的刺激效应,对其中的有效组分作Ｗｅｓｔｅｒｎ ｂｌｏｔｔｉｎｇ等鉴定分析。结果：证实了其中一种为热休克蛋白－７０的膜蛋白能选择性扩增人外周血的γδＴ细胞。结论：表达于人多发性骨髓瘤细胞ＸＧ－７细胞膜上的热休克蛋白－７０多肽是γδＴ细胞     

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。     

空肠弯曲菌HSP43诱导自身抗体产生

小鼠每周注射１次次空弯菌ＨＳＰ４３共１０Ｗ。ＨＳＰ４３抗体第４周起显著升高一直持续到１６Ｗ，抗ｄｓ－ＤＮＡ，ｓｓ－ＤＮＡ自身抗体在６Ｗ和１６Ｗ出现两个高峰，抗ＥＮＡ自身抗体在４－１６Ｗ逐渐升高，抗组蛋白抗体在８Ｗ最高然后下降。     

热休克蛋白60口服耐受对小鼠动脉粥样硬化斑块的影响

目的:探讨热休克蛋白60(HSP60)口服诱导的免疫耐受对ApoE-/-小鼠动脉粥样硬化斑块的影响.方法:8周龄ApoE-/-小鼠分别口服重组鼠HSP60 PBS溶液和单纯PBS,连续5天后高脂饲养12周.观察各组斑块面积;分析脾细胞中CD4+CD25+调节性T细胞百分比;脾细胞抗原特异性T细胞增殖反应,ELISA法检测上清液细胞因子浓度.结果:与对照组比较,口服HSP60组斑块面积显著减小,口服HSP60组脾细胞中CD4+CD25+调节性T细胞百分比显著升高(P<0.01).HSP60刺激脾细胞后,口服HSP60组T细胞增殖反应下降,上清液中TGF-β浓度显著升高,而IFN-γ显著下降.结论:口服HSP60可预防动脉粥样硬化形成;口服HSP60诱导调节性T细胞介导的抗原特异性免疫耐受可能是动脉粥样硬化的机制.     

热休克蛋白的合成调控

无论是原核生物还是真核生物受热及其他应激作用后都能诱导热休克蛋白(HSP)的合成。反应的普遍性反映了生物体的进化保守性,提示了HSP可能的重要生理功能。近年来对于热休克反应机理的研究逐渐增多,对热休克期间基因表达与调控了解较多。本文介绍了热休克基因结构,编码基因,激活因子和有关的协助因子,分别阐述了真核与原核生物不同水平的基因表达调节。虽然这种调节可发生在基因活化及表达的各个层次,但转录阶段似乎是最有效的。     

热休克蛋白22研究进展

A newly identified 22 kD protein, which is named as heat shock protein22 (HSP22), is a member of sHSP (small heat shock protein) subfamily. It contains the α-crystallin domain in its C-terminal half, a hallmark of the subfamily of sHSP. HSP22 has Ser-Thr protein kinase activity and chaperone-like activity. HSP22 is expressed in multiple tissues, specially in skeletal muscle, placenta and heart. Previous studies showed that HSP22 involved in growth and transformation of cells, apoptosis of tumor, hypertrophy and apoptosis of myocardial cells.     

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     

热休克蛋白70对荷瘤鼠的治疗作用

目的研究肿瘤来源的热休克蛋白70(heat shock protein 70,HSP70)对荷瘤小鼠的治疗作用,为其应用提供参考依据.方法细胞培养、蛋白质提纯技术、western-blot法、毛细管电泳技术和动物实验等.结果应用5μgHSP70能延长小鼠的生存期限,平均生存(28.7±4.5)d,与对照组(18.5±3.9)d相比差异显著(P＜0.05);而10μg治疗组的小鼠生存期为＞(62.3±29.1)d,40%获长期生存(＞90d),所接种肿瘤完全消退.与其他组相比,差异具显著性(P＜0.01).结论肿瘤来源的HSP70具有明确的治疗作用,本研究对于研究应用肿瘤来源的热休克蛋白70治疗人类恶性肿瘤具有较重要的参考意义.     

热休克蛋白在乳腺癌中的作用

热休克蛋白是细胞受各种应激原刺激后诱导产生的一组应激蛋白,具有维持细胞蛋白稳定、促进细胞生存等功能,在细胞生长、发育、分化、基因转录等方面发挥重要作用.近年来发现热休克蛋白也与肿瘤细胞的发生、发展密切相关.在多种肿瘤组织中发现热休克蛋白存在高表达,尤其在乳腺癌中与患者预后密切相关.本文就热休克蛋白的结构、生物学功能、致癌机理以及与乳腺癌的关系作一概述.     

天花粉蛋白合成肽通过激活抑制性CD8~+ T细胞诱导免疫抑制

为了探讨天花粉蛋白合成肽(M-Tk)诱导免疫抑制的机制,应用小鼠针对可溶性抗原OVA的增殖系统,检测M-Tk处理后对淋巴细胞增殖的抑制作用和细胞因子格局的变化,以及M-Tk激活的T淋巴细胞亚群的表型及功能。结果表明,M-Tk可诱导一群CD8+ T抑制性细胞,其表型为CD8+CD28-CTLA4+,采用双层非接触共培养系统发现,该群抑制性细胞主要依赖于细胞因子IL-10、IL-4和细胞间接触发挥免疫抑制作用。     

Interleukin-2 directly induces activation and proliferation of chicken T cells in vivo.

Cytokines, as immune activators, have been investigated in mammalian systems as natural adjuvants and therapeutics. In particular, interleukin-2 (IL-2) has been studied widely as a vaccine adjuvant and immuno-enhancer because of its role in activating T cell proliferation. We show here that the first nonmammalian IL-2 gene cloned, chicken IL-2 (ChIL-2), exhibits similar biologic activities to those of mammalian IL-2. To assess the activities of ChIL-2 in vivo, we injected birds with recombinant ChIL-2 (rChIL-2) protein. rChIL-2 treatment induced peripheral blood lymphocytes to express cell surface IL-2 receptors (IL-2R) within 48 h and resulted in an increase in the proportion of peripheral blood CD4+ and CD8+ T cells. Using bromodeoxyuridine (BrdU) incorporation as a measurement of cell proliferation, we showed the increase in T cell populations to be due to cell proliferation. The ability of ChIL-2 to cause both activation and proliferation of T cells in vivo indicates that it has the potential to be used as an immune activator.     

Campylobacter jejuni induces maturation and cytokine production in human dendritic cells

Campylobacter jejuni is a leading bacterial cause of human diarrheal disease in both developed and developing nations. Colonic mucosal invasion and the resulting host inflammatory responses are thought to be the key contributing factors to the dysenteric form of this disease. Dendritic cells (DCs) play an important role in both the innate and adaptive immune responses to microbial infection. In this study, the interaction between human monocyte-derived dendritic cells and C. jejuni was studied. We found that C. jejuni was readily internalized by DCs over a 2-h period. However, after a prolonged infection period (24 or 48 h) with C. jejuni, only a few viable bacteria remained intracellularly. Minimal cytotoxicity of C. jejuni to dendritic cells was observed. C. jejuni induced the maturation of dendritic cells over 24 h, as indicated by up-regulation of cell surface marker proteins CD40, CD80, and CD86. In addition, Campylobacter-infected DCs triggered activation of NF-kappaB and significantly stimulated production of interleukin-1beta (IL-1beta), IL-6, IL-8, IL-10, IL-12, gamma interferon, and tumor necrosis factor alpha (TNF-alpha) compared to uninfected DCs. Active bacterial invasion of DCs was not necessary for the induction of these cytokines, as heat-killed C. jejuni stimulated similar levels of cytokine production as live bacteria. Purified lipooligosaccharide of C. jejuni appears to be the major stimulant for the increased production of cytokines by DCs. Taken together, these data indicate that during infection, Campylobacter triggers an innate inflammatory response through increased production of IL-1beta, IL-6, IL-8, and TNF-alpha and initiates a Th1-polarized adaptive immune response as predicted from the high level of production of IL-12.     

空肠弯曲菌外膜蛋白亚单位疫苗的研制及免疫原性研究

目的 探索用脂质体包裹空肠弯曲菌(Campylobacter jejuni,CJ)Mr 28 000～31 000外膜蛋白制备安全、稳定、有效的CJ亚单位脂质体佐剂疫苗及其免疫原性、免疫剂量和途径.方法 采用薄膜-超声-冻融法制备阴离子脂质体(AL)、阳离子脂质体(CL)CJ疫苗后对其进行质量鉴定;用ELISA(间接法)检测经CJ疫苗免疫后的新西兰兔血清IgG抗体水平.结果 ①疫苗无菌试验及热原质试验均合格,均无明显不良反应;②阴、阳离子脂质体疫苗冻融前包裹率分别为32.1%、22.5%,冻融后分别是38.3%、28.4%;③CJ阴离子组比CJ阳离子组血清IgG的OD值显著增高(P<0.01),与CJ弗氏组无统计学差别(P0.1);CJ阴、阳离子组0.50 mg/kg的免疫剂量血清IgG的OD值最大;CJ阳离子皮下注射组与肌肉注射组IgG的OD值无统计学差别(P0.1);特异性IgG抗体经6个月动态观察,5个月后抗体效价下降.结论 ①采用薄膜-超声-冻融法制备CJ亚单位佐剂疫苗安全、稳定,冻融法可以提高脂质体疫苗包裹率;②同一剂量的阴离子脂质体疫苗血清抗体水平明显高于阳离子脂质体疫苗;③脂质体疫苗最佳接种剂量为0.50 mg/kg,肌肉注射与皮下注射免疫效果无显著差异;④较高水平的特异性抗体维持5个月,6个月后应加强免疫.     

空弯菌抗原诱导的自身抗体产生动力学

用CJ-S131菌苗经口免疫小鼠16周,观察免疫小鼠体内的自身抗体(抗ds-DNA,ss-DNA,组蛋白和ENA抗体)产生动力学。在CJ-S131菌苗连续免疫的16周内,免疫小鼠有两个自身抗体产生峰。免疫后第6周,出现第一峰,抗组蛋白和ENA抗体的阳性率高于抗ds-DNA和ss-DNA抗体;在第16周,抗ds-DNA和ss-DNA抗体的阳性率高于抗组蛋白和ENA抗体。在正常小鼠体内,存在“自然”抗核酸抗体(抗ds-DNA和ss-DNA抗体),随年龄而增长,但个体差异较大;而抗核蛋白抗体(抗组蛋白和ENA抗体)始终测不出。上述结果提示,抗组蛋白抗体和抗ENA抗体在自身免疫病的发病机理和早期诊断中,具有潜在意义,值得进一步研究。     

肿瘤源性热休克蛋白gp96诱导淋巴细胞反应及其抗肿瘤效应

目的：研究肿瘤细胞来源的热休克蛋白gp96-多肽复合物在体外诱导脾淋巴细胞的特异性细胞毒性T淋巴细胞（CTL）反应.方法：利用蛋白纯化技术、SDS-PAGE凝胶电泳及Western blot法分离纯化、鉴定gp96-多肽;通过流式细胞术、免疫荧光技术、CCK-8法等检测经gp96多肽诱导的CD8^＋T细胞及其抗肿瘤效应.结果：经鉴定获得纯化的热休克蛋白;流式细胞仪检测表明,经gp96-肽复合物诱导后的CD8^＋T细胞比例达到近70%,远远高于对照组的35%、26%;该活化的CTL细胞在效靶比为50：1时的肿瘤杀伤率达72%,与对照组相比具有统计学意义;激光共聚焦显微镜观察证实实验诱导组的培养上清能诱导H22肿瘤细胞凋亡的形态学改变.结论：肿瘤来源的热休克蛋白gp96-肽复合物能诱导小鼠脾淋巴细胞的CTL反应,该活化的CTL具有特异性抗H22肿瘤细胞的免疫作用,并能分泌免疫活性物质诱导H22肿瘤细胞凋亡.