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树突状细胞体外刺激对HBV特异性细胞毒T细胞影响的研究 被引量:15
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作者 张恒辉 何豫 +5 位作者 赵鸿 朴文花 刘茂昌 席宏丽 于敏 王贵强 《中华医学杂志》 CAS CSCD 北大核心 2005年第17期1171-1176,共6页
目的探讨通过聚肌胞体外作用树突状细胞后改善慢性乙型肝炎患者树突状细胞功能,并在体外活化自身T细胞获得高频数HBV特异性细胞毒性T细胞(CTL)的方法。方法分离慢性乙型肝炎病人外周血单个核细胞,在粒巨噬细胞集落刺激因子(GM CSF)和白... 目的探讨通过聚肌胞体外作用树突状细胞后改善慢性乙型肝炎患者树突状细胞功能,并在体外活化自身T细胞获得高频数HBV特异性细胞毒性T细胞(CTL)的方法。方法分离慢性乙型肝炎病人外周血单个核细胞,在粒巨噬细胞集落刺激因子(GM CSF)和白细胞介素4(IL4)的诱导下培养树突状细胞。培养的第7天加入聚肌胞刺激获得成熟树突状细胞,经HBVcore1827肽负载后与自身T淋巴细胞共培养,通过酶联斑点计数法(Elispot)及MHC肽四聚体法(Tetramer)比较病人T细胞未经自身树突状细胞刺激组、经HBVcore1827肽负载的自身树突状细胞刺激组、经聚肌胞促成熟的HBVcore1827肽负载的自身树突状细胞刺激组中HBV特异性的CTL的功能和频数。结果慢性乙型肝炎病人外周血单核细胞体外经GM CSF和IL4诱导可转化为树突状细胞,树突状细胞转化过程中聚肌胞的刺激可显著上调树突状细胞表面分子CD80、CD83的表达(P<0.01),促进树突状细胞的成熟。Elispot法检测分泌IFNγ的CTL的频数病人T细胞未经自身树突状细胞刺激组分泌频数为(9~28)/1×105T细胞,均值16;经HBVcore1827肽负载的自身树突状细胞刺激组频数为(30~67)/1×105T细胞,均值为46;经聚肌胞促成熟的HBVcore1827肽负载的自身树突状细胞刺激组频数为(59~130)/1×105T细胞,均值为98。三组数据? 展开更多
关键词 特异性细胞毒T细胞 影响的研究 白细胞介素-4(IL-4) 自身树突状细胞 体外刺激 慢性乙型肝炎患者 巨噬细胞集落刺激因子 慢性乙型肝炎病 HBV特异性 人外周血单个核细胞 Elispot法 tetramer 外周血T淋巴细胞 细胞毒性T细胞
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加载HCMV抗原肽的HLA-A*0201单体及其四聚体制备和鉴定 被引量:9
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作者 何贤辉 徐丽慧 +2 位作者 刘毅 蔡小嫦 曾耀英 《生物工程学报》 CAS CSCD 北大核心 2004年第3期382-388,共7页
细胞毒T淋巴细胞 (CTL)在控制病原体感染以及抗肿瘤过程中发挥重要作用 ,因而特异性CTL的检测相当重要 ;而过去检测CTL的方法都是间接的 ,最近发展起来的四聚体技术则是直接检测抗原特异性CTL的有效而特异的方法 ,成为目前研究T细胞免... 细胞毒T淋巴细胞 (CTL)在控制病原体感染以及抗肿瘤过程中发挥重要作用 ,因而特异性CTL的检测相当重要 ;而过去检测CTL的方法都是间接的 ,最近发展起来的四聚体技术则是直接检测抗原特异性CTL的有效而特异的方法 ,成为目前研究T细胞免疫应答的关键技术。报道一种简化的四聚体制备程序 ,利用该程序成功制备加载人巨细胞病毒 (HCMV)抗原肽的HLA_A2四聚体 ,具有特异性结合CTL活性。HLA_A 0 2 0 1重链基因是通过RT_PCR方法从HLA_A2 + 供者白细胞中克隆 ,进而以PCR方法构建在羧基端融合生物素化酶BirA底物肽 (BSP)的HLA_A 0 2 0 1(A2 )重链胞外区原核表达载体 ,A2重组蛋白在大肠杆菌中得到高表达 ,主要以包涵体形式存在。加载抗原肽的可溶性A2单体是A2胞外区在轻链 β2 微球蛋白和HLA_A2限制性HCMVpp6 5 495_50 3抗原肽 (NLVPMVATV ,NLV)存在时通过稀释法复性获得 ,以BirA对其进行生物素化 ,然后以阴离子交换树脂纯化 ,得到的纯化A2_NLV单体与Streptavidin_PE按 4 :0 8比例混合形成四聚体 ,结合程度在 85 %以上 ,流式细胞仪分析显示该四聚体具有与HLA_A2 + 供者的特异性CTL结合活性。总之 ,这种简化的四聚体制备程序 ,不仅有利于该技术的推广 ,为特异性T细胞免疫研究建立必要的技术平台 。 展开更多
关键词 人类白细胞抗原 四聚体 细胞毒T淋巴细胞 包涵体 人巨细胞病毒
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油溶性四元聚合物MASM稠油降粘剂的合成研究 被引量:8
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作者 杨思妍 陈馥 +1 位作者 周慧 高翔 《应用化工》 CAS CSCD 2011年第2期299-302,共4页
设计合成了一种主要针对稠油中胶质沥青质结构特点的四元共聚物降粘剂,对合成聚合物的单体配比、引发剂用量、聚合反应时间、聚合反应温度以及酯化温度进行了研究。通过红外的表征,证实得到了预期的分子结构。确定了最佳合成条件:甲基... 设计合成了一种主要针对稠油中胶质沥青质结构特点的四元共聚物降粘剂,对合成聚合物的单体配比、引发剂用量、聚合反应时间、聚合反应温度以及酯化温度进行了研究。通过红外的表征,证实得到了预期的分子结构。确定了最佳合成条件:甲基丙烯酸甲酯、马来酸酐、苯乙烯、丙烯酰胺的摩尔比为10∶2∶3∶1,甲苯为溶剂,偶氮二异丁晴作引发剂,引发剂用量为单体重10‰,在73℃下反应6 h所得到的共聚物,加入十八醇(酯醇比为1∶1.2)在85℃下酯化4 h,得到降粘率将近90%的聚合物。 展开更多
关键词 油溶性降粘剂 稠油 四元共聚 性能评价
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一种油溶性稠油降粘剂的制备与性能评价 被引量:8
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作者 任昊 《广州化工》 CAS 2014年第20期89-90,136,共3页
以丙烯酸十八酯与丙烯酰胺、马来酸酐单体与一种阴离子单体对乙烯基苯磺酸钠(NaSS)通过溶液聚合的方法合成了一种油溶性聚合物。结果表明:所合成的降粘剂对塔河JT-4稠油具有良好的降粘效果。降粘剂加入量为400 mg/kg时,该降粘剂在50℃... 以丙烯酸十八酯与丙烯酰胺、马来酸酐单体与一种阴离子单体对乙烯基苯磺酸钠(NaSS)通过溶液聚合的方法合成了一种油溶性聚合物。结果表明:所合成的降粘剂对塔河JT-4稠油具有良好的降粘效果。降粘剂加入量为400 mg/kg时,该降粘剂在50℃使高粘稠油的降粘率几乎达到90%,具有极好的降粘效果。 展开更多
关键词 稠油 油溶性降粘剂 丙烯酸十八酯 马来酸酐 四元共聚
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A New Isopropenyl Benzofuran-type Tetramer from Ligularia stenocephala 被引量:2
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作者 Fu Lin YAN Ai Xia WANG +1 位作者 Zhong Jian JIA Lei HE 《Chinese Chemical Letters》 SCIE CAS CSCD 2003年第12期1253-1254,共2页
A new isopropenyl benzofuran-type tetramer was isolated from the roots of ligularia stenocephala and its structure was established by spectroscopic methods.
关键词 COMPOSITAE Ligularia stenocephala isopropenyl benzofuran-type tetramer.
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Construction of Soluble Mamu-B~*1703,a Class I Major Histocompatibility Complex of Chinese Rhesus Macaques,Monomer and Tetramer Loaded with a Simian Immunodeficiency Virus Peptide 被引量:4
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作者 Dongyun Ouyang Xiaoying Wang +4 位作者 Xianhui He Lihui Xu Huanjing Shi Qi Gao He Guo 《Cellular & Molecular Immunology》 SCIE CAS CSCD 2009年第2期117-122,共6页
Chinese-descent rhesus macaques have become more prevalent for HIV infection and vaccine investigation than Indian-origin macaques. Most of the currently available data and reagents such as major histocompatibility co... Chinese-descent rhesus macaques have become more prevalent for HIV infection and vaccine investigation than Indian-origin macaques. Most of the currently available data and reagents such as major histocompatibility complex (MHC) class I tetramers, however, were derived from Indian-origin macaques due to the dominant use of these animals in history. Although there are significant differences in the immunogenetic background between the two macaque populations, they share a few of common MHC class I alleles. We reported in this study the procedure for preparation of a soluble Mamu-B*1703 (a MHC class I molecule of Chinese macaques) monomer and tetramer loaded with a dominant simian immunodeficiency virus (SIV) epitope IW9 (IRYPKTFGW) that was identified to be Mamu-B*1701-restricted in Indian macaques. The DNA fragment encoding the Mamu-B*1703 extracellular domain fused with a BirA substrate peptide (BSP) was amplified from a previously cloned cDNA and inserted into a prokaratic expression vector. In the presence of the antigenic peptide IW9 and light chain β2-microglobulin, the expressed heavy chain was refolded into a soluble monomer. After biotinylation, four monomers were polymerized as a tetramer by phycoerythrin-conjugated streptavidin. The tetramer, having been confirmed to have the right conformation, was a potential tool for investigation of antigen-specific CD8^+ T-lymphocytes in SIV vaccine models of Chinese macaques. And our results also suggested that some antigenic peptides reported in Indian-origin macaques could be directly recruited as ligands for construction of Chinese macaque MHC tetramers. 展开更多
关键词 Mamu-B* 17 MHC class I rhesus macaque tetramer
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草鱼免疫球蛋白IgM四聚体的分析鉴定 被引量:6
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作者 陈丛琳 罗绍祥 +1 位作者 张晓华 戴和平 《水生生物学报》 CAS CSCD 北大核心 2012年第1期173-176,共4页
草鱼(Ctenopharyngodonidellus)是我国重要的淡水经济鱼类之一。为了制备有效的抗各种病原体的疫苗,必须对草鱼的免疫应答分子进行分析。IgM是大多数真骨鱼类血清中都存在的一种免疫球蛋白,是主要的介导体液免疫应答的分子,也是有... 草鱼(Ctenopharyngodonidellus)是我国重要的淡水经济鱼类之一。为了制备有效的抗各种病原体的疫苗,必须对草鱼的免疫应答分子进行分析。IgM是大多数真骨鱼类血清中都存在的一种免疫球蛋白,是主要的介导体液免疫应答的分子,也是有颌类脊椎动物进化过程中最先出现的免疫球蛋白。 展开更多
关键词 草鱼 IGM 四聚体
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Construction and Functional Test of HLA-A*2402-Peptide Tetramer 被引量:3
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作者 XiaolingLu XiongwenWu +3 位作者 ZhihuiLiang XiufangWeng QingLi FeiliGong 《Cellular & Molecular Immunology》 SCIE CAS CSCD 2005年第2期145-149,共5页
HLA-A~*2402 is one of the most frequent HLA-A allele in Asian population.To construct HLA-A~*2402-peptide tetramers,the transmembrane and intracellular segments of HLA-A~*2402 cDNA were replaced with BSP sequence to f... HLA-A~*2402 is one of the most frequent HLA-A allele in Asian population.To construct HLA-A~*2402-peptide tetramers,the transmembrane and intracellular segments of HLA-A~*2402 cDNA were replaced with BSP sequence to form a fusion gene of sHLA-A~*2402-BSP.The sHLA-A~*2402-BSP fusion protein and β2m were high-level expressed as insoluble aggregates in E.coli,and refoided to form an HLA-A~*2402-peptide monomeric complex by dilution method in the presence of an antigenic peptide.The HLA-A~*2402-peptide monomeric complex was biotinated and tetramized to prepare HLA-A~*2402-peptide tetramer.Then using the HLA-A~*2402-peptide tetramers to detect antigen-specific cytotoxic T lymphocyte(CTL)induced by artificial antigen presenting cell (aAPC)in vitro.The results showed that HLA-A~*2402-peptide tetramer was prepared correctly,and functional in detecting antigen-specific CTL in vitro,HLA-A~*2402-peptide monomeric and its multimeric complexes are expected to provide a powerful tool for studying mechanisms of immune-related diseases in Asian populations. Cellular & Molecular Immunology.2005;2(2):145-149. 展开更多
关键词 tetramer HLA-A~*2402-peptide monomeric complex CTL
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A New Stilbene Tetramer from Caragana rosea 被引量:2
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作者 Guo Xun YANG, Chang Qi HU School of Pharmacy, Fudan University, Shanghai 200032 《Chinese Chemical Letters》 SCIE CAS CSCD 2003年第10期1048-1050,共3页
Cararosinol A, a new stilbene tetramer, was isolated from Caragana rosea. Its structure has been established on the basis of spectroscopic evidence.
关键词 Caragana rosea Leguminase stilbene tetramer.
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OVA_(257-264)肽-β2m融合基因的构建及体内特异性CTL的诱导 被引量:4
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作者 钱丽 钱关祥 《细胞与分子免疫学杂志》 CAS CSCD 北大核心 2002年第5期439-442,446,共5页
目的 构建融合基因OVA2 57 2 64 β2m的表达载体 ,并以其表达产物作为免疫原 ,在体内诱导特异性CTL。方法 采用PCR技术 ,从人肝癌细胞株SMMC 772 1中克隆人β2m基因 ,拼接上OVA(2 5 7~ 2 6 4)序列及linker后 ,克隆入pET 42b(+)高效... 目的 构建融合基因OVA2 57 2 64 β2m的表达载体 ,并以其表达产物作为免疫原 ,在体内诱导特异性CTL。方法 采用PCR技术 ,从人肝癌细胞株SMMC 772 1中克隆人β2m基因 ,拼接上OVA(2 5 7~ 2 6 4)序列及linker后 ,克隆入pET 42b(+)高效表达载体进行诱导表达。对表达产物进行纯化与复性后 ,用其作为免疫原诱导特异性CTL ,并构建H 2Kb OVA四聚体检测特异性CTL。结果 成功地获得融合蛋白OVA2 57 2 64 β 2m及H 2Kb OVA四聚体 ,应用H 2Kb OVA四聚体对特异性CTL检测结果与经典的细胞毒试验一致。结论 OVA2 57 2 64 β 2m的表达产物 ,可有效的在体内诱导特异性CTL应答。在体外构建的MHCI类分子四聚体 。 展开更多
关键词 四聚体 表达 纯化 特异性CTL PCR技术 多肽疫苗
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大鼠金属硫蛋白晶体堆积形式的理论研究 被引量:5
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作者 侯廷军 安钰 +1 位作者 茹炳根 徐筱杰 《高等学校化学学报》 SCIE EI CAS CSCD 北大核心 2002年第2期271-274,共4页
考虑到蛋白质在结晶过程中溶液和晶体之间的动态平衡 ,蛋白质在溶液中的聚合状态决定了晶体堆积的最小单元 .通过动态光散射实验以及计算机模拟 ,对大鼠金属硫蛋白亚型 和兔肝金属硫蛋白亚型 和 的聚合物形式进行了比较 ,可以确定大... 考虑到蛋白质在结晶过程中溶液和晶体之间的动态平衡 ,蛋白质在溶液中的聚合状态决定了晶体堆积的最小单元 .通过动态光散射实验以及计算机模拟 ,对大鼠金属硫蛋白亚型 和兔肝金属硫蛋白亚型 和 的聚合物形式进行了比较 ,可以确定大鼠金属硫蛋白亚型 在结晶条件下的缓冲液中应该以四聚体而不是二聚体形式存在 .通过考察晶体的模型也可以发现 ,晶体的确可以通过四聚体的形式堆积得到 。 展开更多
关键词 金属硫蛋白晶体 二聚体 三聚体 四聚体 X射线晶体学 大鼠 堆积形式 结构测定
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猪β2m四聚体前体链重组表达载体的构建和表达 被引量:5
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作者 刘昌华 刘筏 +2 位作者 张晶 乔震 高凤山 《中国畜牧兽医》 CAS 北大核心 2013年第4期77-81,共5页
为构建猪β2m轻链四聚体前体链,通过PCR定点突变技术,将猪主要组织相容性复合体Ⅰ类分子轻链β2m羧基端69位苯丙氨酸(Phe)突变为半胱氨酸(Cys),然后将突变体基因插入到pET-28a载体,转化BL21细胞,经诱导后SDS-PAGE检测蛋白质表达情况。... 为构建猪β2m轻链四聚体前体链,通过PCR定点突变技术,将猪主要组织相容性复合体Ⅰ类分子轻链β2m羧基端69位苯丙氨酸(Phe)突变为半胱氨酸(Cys),然后将突变体基因插入到pET-28a载体,转化BL21细胞,经诱导后SDS-PAGE检测蛋白质表达情况。结果显示,β2m突变体基因全长297bp,编码98个氨基酸,其中羧基端69位Phe突变为Cys。突变体基因经诱导表达后进行SDS-PAGE检测,结果显示蛋白质表达大小为10.6ku。本研究构建了猪β2m轻链突变体基因重组表达载体,并成功表达了目的蛋白,为今后构建四聚体链奠定基础。 展开更多
关键词 β2m基因 四聚体 定点突变 表达
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Hydrothermal Assembly and Structural Characterization of an Octamolybdate Supported Copper (Ⅰ) Tetramer: [Cu_2(1,10-phen)_2(4,4′-bpy)]_2[Mo_8O_(26)]·4H_2O 被引量:2
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作者 吴传德 詹晓平 《Chinese Journal of Structural Chemistry》 SCIE CAS CSCD 北大核心 2002年第5期525-529,共5页
A hydrothermal reaction of copper acetate with ammonium molybdate, 4,4-bpy (4,4-bipyridine) and 1,10-phen (1,10-phenanthroline) led to the formation of brown crystals of [Cu2(1,10-phen)2(4,4-bpy)]2 [Mo8O26]4H2O 1. Si... A hydrothermal reaction of copper acetate with ammonium molybdate, 4,4-bpy (4,4-bipyridine) and 1,10-phen (1,10-phenanthroline) led to the formation of brown crystals of [Cu2(1,10-phen)2(4,4-bpy)]2 [Mo8O26]4H2O 1. Single-crystal X-ray analysis has revealed that 1 (C68H56N12O30Cu4Mo8) crystallizes in the triclinic system, space group P with a = 11.270(3), b = 13.113(6), c = 13.906(3) ? = 103.33(4), b = 98.54(2), g = 101.29(2)? V = 1920.1(1) ?3, Mr = 2542.93, Z = 1, Dc = 2.199 g/cm3, m = 2.435 mm-1, F(000) = 1240, the final R = 0.0445, wR = 0.1082 and S = 1.021 for 5052 observed reflections with I >2s(I). It consists of copper (Ⅰ) tetramer units and -[Mo8O26]4- anions, which are further attached into a three-dimensional framework through hydrogen bonding and - stacking interactions. 展开更多
关键词 heteropolymolybdate crystal structure OCTAMOLYBDATE copper tetramer
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Preparation and Characterization of HLA-A*0201 Tetramer Loaded with IE-1_(316-324) Antigenic Peptide of Human Cytomegalovirus 被引量:1
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作者 Lihui Xu Qingbing Zha +3 位作者 Hong Sun Qiantao Jia Fengyao Li Xianhui He 《Cellular & Molecular Immunology》 SCIE CAS CSCD 2006年第5期367-371,共5页
Major histocompatibility complex (MHC) class I tetramer technology has become the central technique for analyzing antigen-specific CD8^+ T cell responses and it has been widely used to explore the differentiation a... Major histocompatibility complex (MHC) class I tetramer technology has become the central technique for analyzing antigen-specific CD8^+ T cell responses and it has been widely used to explore the differentiation and formation of memory CD8^+ T cells. Previously, a simplified and efficient procedure for preparing high quality HLA-A*0201 tetramers has been established in our lab and the tetramers loaded with HCMV peptide pp6549s.50a has been successfully applied to investigate HCMV-specific CD8^+ T cells in Chinese populations. Using similar procedure we reported here the construction of HLA-A*0201 tetramer loaded with another dominant epitope derived from immediate early (IE)-1 316.324 (VLEETSVML, VLE) of HCMV (A2-VLE) and characterization of this tetramer. After A2-VLE monomer was prepared and purified, its tetramer was then formed at a yield of 83%. The optimized amount of A2-VLE tetramer for staining 100 μl whole blood was 0.5 μg with incubation at 4℃ for 1 h. Furthermore, the dissociation constant of the tetramer binding to the specific CD8^+ T cells of one HLA-A2^+ donor was estimated to be 32.7 nmol/L, which is markedly higher than that of MHC monomer. The construction of A2-VLE tetramer provides an alternative choice for investigating HCMV-specific CD8^+ T cell responses and will deepen our understanding of the differentiation and formation of HCMV-specific memory CD8^+ T cells. Cellular & Molecular Immunology. 展开更多
关键词 HLA tetramer CYTOMEGALOVIRUS immediate early-1 dissociation constant
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HIV/HCV共感染患者病毒特异性CTL细胞定量研究 被引量:4
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作者 张永宏 陈新月 +3 位作者 杨宇 蒋岩 张彤 吴昊 《中华微生物学和免疫学杂志》 CAS CSCD 北大核心 2004年第12期981-984,共4页
目的探讨病毒特异性CTL对HIVHCV共感染患者病情进展的影响机制。方法观察对象为HIVHCV共感染患者、单纯HIV感染者、单纯HCV感染者。采用四聚体技术,运用流式细胞仪检测病毒特异性CTL。前瞻性比较HIV、HCV特异性CTL在三组中的异同,并对HI... 目的探讨病毒特异性CTL对HIVHCV共感染患者病情进展的影响机制。方法观察对象为HIVHCV共感染患者、单纯HIV感染者、单纯HCV感染者。采用四聚体技术,运用流式细胞仪检测病毒特异性CTL。前瞻性比较HIV、HCV特异性CTL在三组中的异同,并对HIVHCV共感染患者的HIV、HCV特异性CTL进行相关性分析。结果HIVHCV共感染组与单纯HIV感染组比较HIV特异性CTL,差异无显著性(P=0.586)。HIVHCV共感染组中HCV特异性CTL的百分数及绝对计数(0.37±0.29,3.52±3.79)均高于单纯HCV感染组(0.15±0.05,0.86±0.33),差异有统计学意义(P=0.001,P=0.002)。HIVHCV共感染组中的HIV特异性CTL与HCV特异性CTL存在正性线性相关(P<0.001),方程成立。并且各系数均有统计学意义,方程似然比(r2)0.761。结论HCV特异性CTL可能是HIVHCV共感染组中肝脏功能损伤加重的原因之一;HIV与HCV在同一患者存在相互影响。 展开更多
关键词 HCV感染 HIV 患者 特异性CTL 共感染 病毒特异性 细胞 性比 四聚体 定量研究
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乙型肝炎患者外周血病毒特异性CD8阳性细胞毒性T淋巴细胞的数量和功能分析 被引量:4
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作者 范振平 施明 +3 位作者 徐东平 张文瑾 张玲霞 王福生 《肝脏》 2012年第1期11-16,共6页
目的分析急性乙型肝炎(AHB)和慢性乙型肝炎(CHB)患者外周血中乙型肝炎病毒(HBV)特异性细胞毒性T淋巴细胞(CTL)的数量及功能。方法 36例HLA-A2阳性的乙型肝炎患者,其中AHB12例,CHB24例。分别用含HBV抗原C、S和P区的c18-27、s183-191和p57... 目的分析急性乙型肝炎(AHB)和慢性乙型肝炎(CHB)患者外周血中乙型肝炎病毒(HBV)特异性细胞毒性T淋巴细胞(CTL)的数量及功能。方法 36例HLA-A2阳性的乙型肝炎患者,其中AHB12例,CHB24例。分别用含HBV抗原C、S和P区的c18-27、s183-191和p575-583三个肽段的四聚体[Tetramer(Tc18-27、Ts183-191、Tp575-583)]检测患者外周血单核淋巴细胞(PBMCs)中HBV特异性CD8阳性CTL细胞的数量,同时用酶联免疫斑点法(ELISPOT)检测其分泌IFN-γ的功能。用SPSS13.0进行统计学分析。结果 AHB和CHB患者外周血中Tc18-27特异性的CTL数量无明显不同;而Ts183-191特异性CTL的平均值分别为0.24%±0.39%和0.03%±0.02%,阳性率分别为75%和33.3%;Tp575-583特异性CTL平均值分别为0.08%±0.09%和0.02%±0.01%,阳性率分别为50%和16.7%,AHB较CHB显著升高(P<0.05)。此外,AHB患者平均Tetramer阳性的个数为1.58个,而CHB患者平均为0.67个,AHB较CHB显著增多(P<0.01)。在9例AHB患者中,其外周血Ts183-191特异性的CTL细胞的数量为139~21735个/106PBMCs,用ELISPOT方法检测相对应的分泌IFN-γ的斑点形成细胞(SFC)为0~252个/106PBMCs,AHB患者Tetramer细胞数和ELISPOT检测的IFN-γ斑点数有明显相关性(P<0.01),而CHB患者则无此相关性。结论 AHB与CHB患者外周血中HBV特异性CTL的数量和分泌IFN-γ的差异提示CTL可能在清除病毒方面发挥着至关重要的作用,是今后慢性乙型肝炎免疫治疗的重要研究方向之一。 展开更多
关键词 病毒性肝炎 乙型 细胞毒性T淋巴细胞 四聚体 酶联免疫斑点法
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sHLA-A*020-1抗原肽复合物的构建 被引量:3
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作者 陈浩 吴雄文 +4 位作者 梁智辉 翁秀芳 韩军艳 黄亚非 龚非力 《现代免疫学》 CAS CSCD 北大核心 2004年第3期232-234,共3页
以原核表达的sHLA A 0 2 0 1 BSP为重链 ,β2 m为轻链 ,与人工合成的EBV抗原肽LMP2A (4 2 6 4 34、NH2 CLGGLLTMV COOH )利用稀释法进行共折叠复性 ,形成可生物素化的可溶性HLA A 0 2 0 1抗原肽复合物。并利用仅与天然构象HLAI类分... 以原核表达的sHLA A 0 2 0 1 BSP为重链 ,β2 m为轻链 ,与人工合成的EBV抗原肽LMP2A (4 2 6 4 34、NH2 CLGGLLTMV COOH )利用稀释法进行共折叠复性 ,形成可生物素化的可溶性HLA A 0 2 0 1抗原肽复合物。并利用仅与天然构象HLAI类分子结合的单抗W6 / 32 ,通过Dot ELISA和Westernblot对折叠产物的构象进行鉴定 ,证实复性后成功获得了天然构象的sHLA A 0 2 0 1 抗原肽复合物。为进一步组装可溶性HLA/抗原肽四聚体以及制备人工抗原提呈细胞奠定基础。 展开更多
关键词 sHLA-A*0201-抗原肽复合物 稀释复性 四聚体
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Interactions between Transmembrane Helices within Monomers of the Aquaporin AtPIP2;1 Play a Crucial Role in Tetramer Formation 被引量:1
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作者 Yun-Joo Yoo Hyun Kyung Lee +11 位作者 Wonhee Han Dae Heon Kim Myoung Hui Lee Jouhyun Jeon Dong Wook Lee Junho Lee Yongjik Lee Juhun Lee Jin Seok Kim Yunje Cho Jin-Kwan Han Inhwan Hwang 《Molecular Plant》 SCIE CAS CSCD 2016年第7期1004-1017,共14页
Aquaporin (AQP) is a water channel protein found in various subcellular membranes of both prokaryotic and eukaryotic cells. The physiological functions of AQPs have been elucidated in many organisms. However, unders... Aquaporin (AQP) is a water channel protein found in various subcellular membranes of both prokaryotic and eukaryotic cells. The physiological functions of AQPs have been elucidated in many organisms. However, understanding their biogenesis remains elusive, particularly regarding how they assemble into tetramers. Here, we investigated the amino acid residues involved in the tetramer formation of the Arabidopsis plasma membrane AQP AtPIP2;1 using extensive amino acid substitution mutagenesis. The mutant proteins V41A/ E44A, F51A/L52A, F87A/191A, F92A/193A, V95A/Y96A, and H216A/L217A, harboring alanine substitutions in the transmembrane (TM) helices of AtPIP2;1 polymerized into multiple oligomeric complexes with a vari- able number of subunits greater than four. Moreover, these mutant proteins failed to traffic to the plasma membrane, instead of accumulating in the endoplasmic reticulum (ER). Structure-based modeling revealed that these residues are largely involved in interactions between TM helices within monomers. These results suggest that inter-TM interactions occurring both within and between monomers play crucial roles in tetramer formation in the AtPIP2;1 complex. Moreover, the assembly of AtPIP2;1 tetramers is critical for their trafficking from the ER to the plasma membrane, as well as water permeability. 展开更多
关键词 AQUAPORIN AtPIP2 tetramer formation interaction between transmembrane helices
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High Level Expression of HLA-A ~*0203-BSP Fusion Protein in Escherichia coli and Construction of Soluble HLA-A ~*0203 Monomer and Tetramer Loaded with Epstein-Barr Virus Peptide 被引量:1
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作者 Qiantao Jia Lihui Xu +3 位作者 Qingbing Zha Xiaoyun Chi Fengyao Li Xianhui He 《Cellular & Molecular Immunology》 SCIE CAS CSCD 2007年第4期301-308,共8页
Major histocompatibility complex (MHC) tetramer technology is critical for characterization of antigen-specific T cells. In the present study we reported the successful generation of HLA-A*0203 tetramer loaded with... Major histocompatibility complex (MHC) tetramer technology is critical for characterization of antigen-specific T cells. In the present study we reported the successful generation of HLA-A*0203 tetramer loaded with Epstein- Barr virus EBNA3596-604 peptide (SVRDRLARL, SVR). Prokaryotic expression vector for the ectodomain of the heavy chain of HLA-A*0203 fused with a BirA substrate peptide (HLA-A*0203-BSP) was constructed and the expression conditions of the fusion protein in Escherichia coli (E. coli) were optimized. The fusion protein was highly expressed in inclusion bodies within E. coil It was then refolded in the presence of 132-microglobulin and SVR peptide to form a soluble HLA-A*0203-SVR monomer. After biotinylation with BirA, the monomer was purified by anion-exchange chromatography and its purity was up to 95%. The tetramer was then formulated by mixing the biotinylated monomer with streptavidin-PE at a ratio of 4:1. Flow cytometry showed that this tetramer could specifically react with antigen-specific CD8^+ T cells, indicating that it was biologically functional. These results provide a foundation for further characterization of antigen-specific CD8^+ T cells from HLA-A*0203 subjects. 展开更多
关键词 HLA-A*0203 tetramer Epstein-Barr virus prokaryotic expression BIOTINYLATION
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Navigating the dynamic landscape of alpha-synuclein morphology: a review of the physiologically relevant tetrameric conformation 被引量:1
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作者 Heather R.Lucas Ricardo D.Fernández 《Neural Regeneration Research》 SCIE CAS CSCD 2020年第3期407-415,共9页
N-acetylatedα-synuclein(αSyn)has long been established as an intrinsically disordered protein associated with a dysfunctional role in Parkinson’s disease.In recent years,a physiologically relevant,higher order conf... N-acetylatedα-synuclein(αSyn)has long been established as an intrinsically disordered protein associated with a dysfunctional role in Parkinson’s disease.In recent years,a physiologically relevant,higher order conformation has been identified as a helical tetramer that is tailored by buried hydrophobic interactions and is distinctively aggregation resistant.The canonical mechanism by which the tetramer assembles remains elusive.As novel biochemical approaches,computational methods,pioneering purification platforms,and powerful imaging techniques continue to develop,puzzling information that once sparked debate as to the veracity of the tetramer has now shed light upon this new counterpart inαSyn neurobiology.Nuclear magnetic resonance and computational studies on multimericαSyn structure have revealed that the protein folding propensity is controlled by small energy barriers that enable large scale reconfiguration.Alternatively,familial mutations ablate tetramerization and reconfigure polymorphic fibrillization.In this review,we will discuss the dynamic landscape ofαSyn quaternary structure with a focus on the tetrameric conformation. 展开更多
关键词 ALPHA-SYNUCLEIN amyloid FIBRILS intrinsically disordered PROTEIN MULTIMER N-ACETYLATION oligomer Parkinson’s disease PROTEIN folding PROTEIN structure tetramer
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