抗von willebrand因子单克隆抗体的免疫放射分析法及其临床应用

本文应用抗人von Willebrand 因子(vWF)单克隆抗体建立了两位点免疫放射分析法,以定量测定血浆vWF 抗原水平。固相用抗ⅧR:Ag 抗血清或单克隆抗体SZ-34包被,~(125)Ⅰ标记物用单克隆抗体SZ-29制备。最小检出量为10~(-4)U/ml,工     

正常人外周血单个核细胞白细胞介素-2受体荧光定量方法研究(简报)

本文探索了应用荧光分光光度计定量测定正常人外周血单个核细胞白细胞介素-2受体(IL_2R)的方法及其条件。以PHA与ConA诱导单个核细胞IL_2R表达的最适细胞浓度为3×10~6/ml。PHA和ConA最适浓度分别为100μg/ml和10μg/ml,PHA诱导较ConA为优。PHA诱导细胞IL_2R表达24小时达到高峰,尔后逐渐下降。第一抗体(抗—Tac)与第二抗体(荧光抗体)的最适浓度分别为1:50和1:32,两种抗体与单个核细胞孵育时间分别以20和10分钟为好。检测了同一标本不同复管的细胞(1×10~6)IL_2R结合荧光抗体M为(5.22±0.45)×10~(-10),变异     

恶性肿瘤血清sIL—2R检测及意义

白细胞介素2(IL—2)与其受体(IL—2R)结合后,产生一系列的生物学效应,对机体免疫应答和调节起着关键性作用。可溶性白细胞介素2受体(Soluble Interleukin—2 Receptor,sIL—2)与膜IL—2Rα链(即Tac抗原)基因同源,1985年Rubin首次发现。目前多数学者认为,sIL—2R是细胞IL—2Rα链脱落产物,是淋巴细胞活化的标志。可能参与机体     

全血培养花环法检测人外周血淋巴细胞白介素_2受体

白细胞介素_2 (IL_2)与白细胞介素_2受体(IL_2R)相结合是发挥生物学活性的前提,许多免疫病理的改变如免疫功能低下常伴以IL_2产生不足及IL_2R表达能力降低,因此IL_2R的研究近年来引起人们的注目。本组应用全血培养花环法检测人外周血淋巴细胞IL_2R并与人外周血单纯淋巴细胞培养花环法检测IL_2R法相比较,结果满意,报告如下。材料和方法一、材料RPMI-1640培养液、PHA、淋巴细胞分层液、单克隆抗体一抗Tac,二抗羊抗小鼠gG致敏的红细胞。二、方法 (一)PHA诱导IL_2R:于10m(青霉素小瓶     

抗IL-2R单克隆抗体在肾移植高危受者中的应用

抗IL—2R单克隆抗体可以特异性地与活化T细胞表面的IL—2Rα结合,从而竞争性封闭抑制IL—2,是IL—2/IL—2R介导T细胞增殖的强有力的抑制剂。通常静止T细胞不表达IL—2Rα,因而不受抗IL—2R单克隆抗体影响。而抗体依赖性细胞介导的细胞毒作用,在体外可引起受到抗IL—2R单克隆抗体作用的T细胞发生溶解,可能为另一作用机理。其它的机理还包括:下调IL—2R的表达水平,促使受抗IL—2R单克隆抗体作用的IL—2R从T细胞表面脱落等。IL—2R单克隆抗体不影响循环淋巴细胞总数,而且由于IL—2Rα链伸入细胞浆的尾端较短,因此阻断α链并不引起细胞因子信号转导,因此在临床上不会产生由于细胞因子大量释放而引起的“首剂效应”。     

抗白间素-2受体单克隆抗体预防肾移植排斥

法国南特马塞地区的研究人员进行了一项抗白细胞间素-2受体的单克隆抗体与兔抗胸腺细胞球蛋白预防同种异体肾移植排斥的随机对照试验。白细胞间素-2是激活的T淋巴细胞的主要生长因子,与白细胞间素-2受体的Tac链成分反应的抗体能预防动物的同种移植物排斥反应。由于Tac链仅在激活的一小部分淋巴细胞上表达,所以抗白间素-2受体的单克隆抗体在预防移植排斥中的免疫抑制作用比多克隆抗淋巴细胞球蛋白更具有     

PHA激活的人外周血单个核细胞对丝裂原应答反应的特性

我们用植物血凝素(PHA)体外激活正常人外周血单个核细胞(PBMC)作应答细胞,研究丝裂原再刺激时淋巴细胞增生反应和分泌功能的变化。PHA激活3～6d的PBMC对丝裂原再刺激的应答反应与初刺激的应答相比,结果显示:①对PHA再刺激的增生反应强度变化很大,从明显高于到显著低于初次反应,决定于激活的淋巴细胞在体外休止时间的长短;②再次激活的淋巴细胞产生白细胞介素2(IL-2)的量显著高于初次激活的淋巴细胞,尤以体止1d的细胞为最高;③再次激活的淋巴细胞中膜IL-2受体(mIL-2R),又称Tac阳性细胞百分率显著降低和培养上清液中可溶性IL-2受体(sIL-2R)含量明显增高;④美州商陆有丝分裂原(PWM)不能诱导已激活的PBMC分泌IgG。     

鼠麻风杆菌实验感染模型的细胞免疫

鼠麻风杆菌(MLM)腹腔感染BALB/C和C57BL/6小鼠,随着感染时间的延长,鼠脾重量增加,脾带菌量增多,肠系膜病变加剧;鼠脾细胞对伴刀豆蛋白A(Con A)刺激的增殖反应和产生白细胞介素2(IL 2)的能力进行性降低,BALB/C鼠尤为严重。感染90d时在经Con A活化的BALB/C鼠脾细胞表面未检出IL 2受体,同时发现感染鼠脾细胞可以抑制正常鼠脾细胞经Con A刺激产生IL 2和表达IL 2受体的能力。推测感染鼠脾细胞免疫功能低下可能是该菌活化了抑制细胞,继而抑制T淋巴细胞合成和分泌IL 2和表达IL 2受体。     

雷公藤内酯醇对小鼠IL-2产生和IL-2受体表达的影响

目的　探讨雷公藤内酯醇对小鼠脾淋巴细胞IL 2产生和IL 2受体表达的影响。方法应用体外细胞培养和APAAP桥联酶标法检测了雷公藤内酯醇对刀豆蛋白A(ConA)诱导的小鼠脾淋巴细胞白细胞介素 (IL -2 )产生及IL -2受体表达的影响。结果雷公藤内酯醇对IL -2产生无影响 ,但对IL -2受体表达有明显的抑制作用。结论雷公藤内酯醇抗免疫排斥作用主要是通过抑制IL -2受体表达 ,从而使IL -2与IL -2受体的相互作用受到影响 ,最终抑制了淋巴细胞增殖。     

抗HLA单克隆抗体对T细胞增殖反应的影响

本文研究了抗HLA—Ⅰ类及抗HLA—D_R单克隆抗体对T细胞增殖反应的影响。结果发观,抗HLA—T类及抗HLA—D_R单克隆抗体均明显抑制T细胞对刺激细胞及PHA的增殖反应。提示非T细胞上的HLA—Ⅰ类抗原可能也参与混合淋巴细胞反应及辅助T细胞对PHA的增殖反应。     

抗原刺激后小鼠CD4~＋ T细胞IL-10受体Ⅰ表达情况的研究

目的：检测抗原刺激后小鼠CD4＋T淋巴细胞表面白细胞介素10受体Ⅰ（interleutin-10 receptorⅠ,IL-10R1）表达变化情况。方法：取C57BL/6小鼠脾细胞,溶血后采用尼龙毛去除B淋巴细胞后,流式细胞仪无菌分选CD4＋T淋巴细胞,利用anti-CD3和anti-CD28多克隆抗体刺激分选后细胞,分别在第0、12、24、48和72 h应用流式细胞术检测IL-10R1表达情况。结果：0 h时小鼠CD4＋T淋巴细胞不表达IL-10R1,随着刺激时间的延长IL-10R1表达增加,24 h时达到最高,随后表达水平开始下降,72 h时表达情况与0 h基本相同。结论：白细胞介素10（interleutin-10,IL-10）对小鼠CD4＋T淋巴细胞的调节可能是在IL-10R1短暂表达的72 h内。     

IL-12 IL-18对乙型肝炎患者Th_1/Th_2亚群的影响

目的探讨IL-12I、L-18对乙型肝炎患者Th1/Th2亚群的影响。方法常规分离人外周血单个核细胞(PBMC),在培养基中分别加入IL-12、IL-18和植物血凝素(PHA)培养72 h。用ELISA方法检测上清中IFN-γ、IL-4的含量。结果IL-12联合PHA或IL-18联合HA诱导下,乙型肝炎患者的IFN-γ水平明显高于对照组;IL-12联合PHA诱导下,乙型肝炎患者IL-4水平降低;IL-18联合PHA诱导下轻度患者明显降低;IL-12、IL-18联合PHA诱导下,乙肝患者的IFN-γ水平明显高于对照组,IL-4水平有降低趋势。结论IL-12、IL-18能够诱导乙型肝炎患者PBMC中Th2细胞向Th1细胞转化;而且IL-12的体外作用要强于IL-18。     

抗Sm侵入淋巴细胞对分泌IL—2的影响

在 PHA 诱导淋巴细胞产生 IL—2的不同时间用抗Sm 抗体与之反应,结果证实,在诱生前24h或诱生开始时加入抗 Sm,淋巴细胞分泌 IL—2的功能受到显著的抑制,在诱生24h 后加入抗 Sm,则不受影响。     

Effect of insulin on the expression of interleukin 2 receptor

Summary In this paper, we report our work about the effect of insulin on the expression of interleukin 2 receptor (IL-2R). Our experiments showed that insulin was not T cell growth factor, but was able to augment the expression of IL-2R during T cell activation by PHA and to delay the decline rate of IL-2R⁺ cell. As a result, the percentage of IL-2R⁺ cells significantly increased during and after activation. Furthermore, insulin was capable of enhancing the production of interleukin 1 by macrophage and interleukin 2 by T lymphocyte. Based on these results, we suggest that there are at least two related mechanisms involved in the augmentation of the expression of IL-2R on activated T cells, i.e., direct action of insulin on T cells and indirect action of insulin by enhancing the production of interleukin l.     

乙肝疫苗接种后无、弱应答者单个核细胞培养上清IL-2、sIL-2R水平测定

目的 :探讨乙肝疫苗接种后无、弱应答发生原因和机制。方法 :选择 2 1名乙肝疫苗接种后无、弱应答者为研究对象 ,同时设 2 2名强应答者为对照组。采用体外细胞培养和酶联免疫检测技术 ,分离外周血单个核细胞(PBMC) ,用特异性抗原HBsAg和非特异性抗原植物血凝素 (PHA)刺激培养 ,测定细胞培养上清中的白细胞介素 - 2(IL - 2 )及其可溶性受体 (sIL - 2R)水平。结果 :经HBsAg和PHA刺激后 ,无、弱应答组的PBMC培养上清中IL - 2水平明显低于强应答组 (P <0 .0 5 ) ,而sIL - 2R水平 2组间差异无统计学意义。结论 :PBMC分泌IL - 2水平低下可能是乙肝疫苗接种后无、弱应答发生的原因和机制之一。     

Changes of intracellular IP3 with the expression of interleukin 2 receptor in human peripheral blood T lymphocytes

Summary The changes of the intracellular inositol-l,4,5-triphosphate (IP₃) associated with the expression of the interleukin 2 receptor (IL-2R) were studied. In resting lymphocytes, IL-2 did not alter intracellular concentration of IP₃, but Con A caused an increase in IP₃ by 45%. In IL-2 sensitive T cells, which expressed IL-2R by 83 %, the change of intracellular IP₃ was dependent upon IL-2 concentration. The IP₃ increased at IL-2 concentrations of 10 and 50 U/ml, and the maximal response of 60 % was found at the concentration of 50 U/ml. At IL-2 concentration of 100 U/ml no increase in IP₃ was observed. After binding of anti-Tac Me Ab to IL-2R of T lymphocytes the increase in IP₃ at IL-2 concentrations of 10 and 50 U/ml was significantly attenuated. It has been suggested that IL-2 could induce the changes of intracellular IP₃ of the human peripheral blood T cells, which is related to the IL-2 concentrations incubated with T cells and the expression of IL-2R on T lymphocyte.     

肝癌患者外周血T细胞亚群与IL－2R表达的临床研究

应用间接免疫荧光法对１４例肝细胞性肝癌患者围术期外周血Ｔ淋巴细胞亚群及单个核细胞ＩＬ－２Ｒ表达行动态观察。结果表明，肝癌患者ＣＤ＿３￣＋细胞、ＣＤ＿４￣＋细胞降低，ＣＤ＿４￣＋／ＣＤ＿８￣＋比值降低，ＣＤ＿８￣＋细胞百分率增高；ＩＬ－２Ｒ表达明显降低。手术切除肿瘤后第１０天，ＣＤ＿４￣＋细胞与ＣＤ＿４￣＋／ＣＤ＿８￣＋比值开始回升，术后３０天升至术前水平，ＩＬ－２Ｒ表达的变化过程与ＣＤ＿４￣＋细胞基本相同。表明手术切除肿瘤负荷能有效地提高宿主免疫功能。     

p38丝裂素活化蛋白激酶信号转录对IL-1β诱导肾小管细胞转分化的影响

目的　探讨 p38MAPK信号转导途径对IL 1β诱导肾小管上皮细胞转分化的影响及其导致的功能改变。方法　以培养的人近端肾小管上皮细胞 (HK 2 )为研究对象 ,给予IL 1β(10ng/ml)刺激 2 4h ,在阻断实验中加入SB2 0 35 80阻断 p38通路。采用蛋白印记杂交法检测 p38MAPK的磷酸化程度 ;细胞表型特征检测包括细胞形态变化、标志蛋白 (角蛋白、α SMA)的表达及分布、细胞增殖、黏附和移行能力。结果　 (1)IL 1β刺激 6～ 4 8h可使α SMA表达上调 ,刺激 2 4h可使角蛋白表达减弱、α SMA分布紊乱 ,但细胞形态变化不明显 ;(2 )IL 1β在 5min时可使p38激活达高峰 ,较基础水平升高 2～ 3倍 (P <0 0 5 ) ;至 12 0min时 p38则呈持续激活状态 ;(3)p38阻断剂 (SB2 0 35 80 )对HK 2细胞α SMA的基础表达有抑制作用 ,抑制率 37 13% (P <0 0 1) ;同时可明显抑制IL 1β刺激的α SMA表达 ,抑制率 4 7 0 7% (P <0 0 0 1) ;(4)IL 1β及 p38阻断剂并不影响HK 2细胞之间的黏附能力 ,但IL 1β可使细胞的移行能力增强 (P <0 0 1) ,而 p38阻断剂可明显抑制其移行能力。IL 1β并不影响HK 2细胞增殖。结论　IL 1β可以激活肾小管上皮细胞p38/MAPK ,并可使其发生表型转化 ,这一作用部分通过 p38信号通路所介导。     

Role of 57 kDa major antigenic component of Shigella dysenteriae outer membrane proteins in induction of major histocompatibility complex II-restricted T-cell response

BACKGROUND: In the past, many Shigella surface antigens were used to activate both T and B lymphocytes but failed to induce antigen-specific responses in Shigellosis. Our objective was to identify in vitro T-cell components using 57 kDa major antigenic fraction of Shigella dysenteriae 1 (IPC-31) outer membrane proteins (OMPs) in modulating specific T-cell subset responses against Shigellosis. METHODS: Antigen-specific T- and B-cell activation was studied in immunized Balb/c mice against 57 kDa antigen by proliferative responses using [3H]-thymidine incorporation and avidin-biotin complex (ABC) peroxidase staining for CD4, CD8, CD3, CD22, and CD25 followed by IL-2 and IL-4 estimation. Macrophage functional assays for migration inhibition factors (MIF) and superoxide (O2-) anions were also performed against 57 kDa antigen, whole OMPs, and phytohemagglutinin (PHA) stimulation. RESULTS: Greater increase of lymphocyte proliferation was observed after 57 kDa antigen stimulation than post-OMP and -PHA stimulation. Proportionately, CD4+ and CD25+ expression of total CD3+ T-cells was significantly dominant (p >0.05) over CD8+ T-cells. On day 7 of this stimulation, it was found to increase % MIF and O2- anions with decrease of IL-2 leading to activation of MHC-II antigens. Later, on day 28 of immunization, IL-2 levels were more increased than on days 7 and 14 but insignificant with non-immunized mice stimulated with 57 kDa. Levels of IL-2 were also noted with low degree of internalization to its IL-2R receptors rather than to IL-4 receptors. In parallel, expression of CD22 was also recorded higher in this stimulation than in PHA, indicating a T-cell-dependent humoral response. CONCLUSIONS: Our results suggested that 57 kDa major antigenic OMP is immunogenic for MHC II-restricted T-cell response to acquire host defense against Shigella infection.     

Immunological status of patients with obstructive jaundice and immunostimulatory effect of arginine

Summary Sepsis is a major factor in the high mortality and morbidity following diagnostic and therapeutic procedures in patients with obstructive jaundice. The reasons for this increased susceptibility to infection are not fully understood. We therefore observed prospectively changes in immunological status of patients with obstructive jaundice in the perioperative period and studied immunological effects of parioperative arginine therapy. The results showed that there was a significant reduction in interleukin 2 (IL-2) production, interleukin 2 receptor (IL-2R) expression and lymphocyte response to phytohemagglutinin (PHA) mitogen in patients with obstructive jaundice compared with normal controls. After operation, the immune suppression in patients with obstructive jaundice was more significant. Arginine is a known T lymphocyte stimulator. Perioperative supplement with arginine significantly enhanced tne immune function of patients with obstructive jaundice, the mechanism being related to increased IL-2 production and IL-2R expression. This study was supported by Grant No. 39000100 from National Natural Science Foundation of China and a Grant from Science Foundation for Young scientists of the Ministry of Public Heath of China.