瘦素对人类外周血T淋巴细胞增殖作用的影响

目的：探讨瘦素（Leptin）对人外周血T淋巴细胞增殖作用的影响.方法：分离并培养人外周血T淋巴细胞,采用[^3H]TdR掺入试验方法,观察不同浓度Leptin的单独作用及其与PHA的协同作用对人外周血T淋巴细胞增殖作用的影响.结果：不同浓度Leptin的单独作用对人类外周血T淋巴细胞均无刺激增殖作用,但可协同PHA的刺激增殖作用,并在一定浓度范围内呈现剂量依赖性.结论：瘦素可协同PHA刺激人类外周血T淋巴细胞增殖.     

苏木对体外人淋巴细胞增殖的抑制作用

作者观察了苏木水煎液体外对PHA或SAC诱导的人淋巴细胞增殖功能,以及诱生IL-2活性的影响.实验以雷公藤水煎液为对照.结果表明苏木体外对SAC诱导的人B淋巴细胞增殖有明显的抑制作用,对PHA诱导的人T淋巴细胞增殖和诱生的IL-2活性亦有明显的抑制作用(P<0.001),其结果与雷公藤相似.而苏木预育人淋巴细胞6h后,则可明显抑制T细胞增殖(P<0.001)和诱生IL-2活性(P<0.05),其抑制强度要明显大于雷公藤(P<0.001).实验提示苏木可作为免疫抑制性中药作进一步的研究和开发.     

小鼠闭合性创伤对脾淋巴细胞增殖活性的影响

本文采用闭合性创伤模型,观察了单纯创伤因素对小鼠脾淋巴细胞增殖活性的影响。结果显示,伤后脾淋巴细胞自发母细胞转化(SBT)活性增强,而分裂原刺激的母细胞转化(MSBT)活性相应降低。提示无外源性感染的单纯创伤对淋巴细胞功能存明显的影响。     

茴香霉素在小鼠淋巴细胞增殖和活化中的作用

目的研究茴香霉素在小鼠淋巴细胞增殖和活化中的作用。方法以活体染料羧基荧光素乙酰乙酸琥珀酰亚胺酯染色，建立了在多克隆刺激剂刀豆蛋白A（ConA）刺激下评价小鼠淋巴细胞增殖的模型，通过流式细胞术及MTT法分析茴香霉素在不同剂量下对淋巴细胞增殖的作用；采用碘化丙锭染色分析茴香霉素对ConA或佛波醇酯（PDB）加离子霉素（Ion）刺激的小鼠淋巴细胞周期变化的作用；利用荧光标记的单克隆抗体双染技术结合流式细胞仪检测茴香霉素对小鼠CD3^＋淋巴细胞早期及中期活化标志分子CD69和CIY25表达的影响。结果随着茴香霉素浓度从1．0ng／ml逐渐增至25．0ng，ml，ConA对淋巴细胞的促增殖作用逐渐减弱，以10．0ng，ml和25．0ng，ml茴香霉素的抑制作用最为明显，呈剂量依赖关系（r=-0．96，P〈0．01）。进一步发现，1．0—25．0ng/ml茴香霉素能够使ConA或PDB加Ion诱导的淋巴细胞停滞于G0/G1期，阻止其进入S期，且阻止作用随上述浓度的增加而增强，呈明显剂量依赖关系（r=-0．97，P〈0．01和r=-0．98，P〈0．01）。据此选用最佳剂量10．0ng/ml，茴香霉素能够明显抑制淋巴细胞表面分子CD69和CIY25的表达（P均〈0．01）。结论茴香霉素能明显抑制小鼠淋巴细胞的增殖和活化。     

CRF患者红细胞促外周血淋巴细胞增殖反应的实验研究

目的 :探讨慢性肾功能衰竭 (CRF)患者红细胞 (RBC)促外周血淋巴细胞 (PBL)增殖能力 ,了解CRF患者RBC免疫调控功能状况。方法 :CRF患者 (检测组 )和正常人 (对照组 )的RBC为效应细胞 ,正常人淋巴细胞为靶细胞 ,采用四甲基偶氮唑蓝 (MTT)比色法 ,检测红细胞调控淋巴细胞增殖反应的能力。结果 :健康人RBC促PBL增殖的作用明显 ,其增殖率为5 7 3%± 10 2 % ,而CRF患者RBC促PBL增殖作用较健康人明显低下 ,增殖率为 32 7%± 7 8% (P <0 0 0 1) ,透析后有较明显的改善 ,增殖率为 4 0 6 %± 11 6 % ,但仍低于健康人。结论 :CRF患者红细胞免疫调控功能低下。     

人胚胸腺激素抑制组分对成人淋巴细胞增殖的影响

在研究人胚胸腺素的基础上,通过对提取工艺的改革,获得了人胚胸腺素抑制组分及促进组分。我们用~3H—TdR掺入DNA法观察了人胚胸腺素抑制组分对成人淋巴细胞增殖的影响,试验结果表明,呈现明显的剂量依赖关系,即40μg的人胚胸腺素抑制组分对PHA诱导的淋巴细胞增殖有显著的抑制作用(P<0.001),当稀释至2μg或更低浓度时,淋巴细胞的增殖恢复到正常范围。而正常淋转无论是高浓度或稀释到较低浓度,都未显示出明显的抑制或促进作用。     

IL-12对慢性丙型肝炎病毒感染者淋巴细胞增殖的影响

目的：观察重组人白细胞介素１２（ｒｈＩＬ－１２）对慢性丙型肝炎病毒（ＨＣＶ）感染者淋巴细胞增殖的影响。方法：外周血单个核细胞（ＰＢＭＣ）分别与植物血浆凝素（ＰＨＡＳ）,ＨＣＶ抗的ｅ２２,ｅ１００－３、ｒｈＩＬ－１２和抗ＩＬ－１２抗体孵育５天后,加入胸腺嘧啶核苷（＾３Ｈ－ＴｄＲ）,然后收集细胞于液闪仪测定每分钟脉冲数（ｃｐｍ）。结果：不论是健康对照者（Ｐ〈０．０２５）,抑或慢性ＨＣＶ感染者（Ｐ〈０．     

应激对淋巴细胞增殖抑制机制的初探

本文在观察了应激小鼠体内抑制因子对淋巴细胞增殖抑制作用的基础上，从应激小鼠血清对淋巴细胞钙离子通道，细胞凋亡及ｃ－ｆｏｓ基因表达影响的３个方面，对其机制进行初步探讨，结果表明，应激小鼠血清可显著抑制ＣｏｎＡ激活的淋巴细胞钙离子通道开放，对淋巴细胞发生凋亡及ｃ－ｆｏｓ基因的表达无影响，本结果提示，抑制淋巴细胞钙离子通道开放，可能是应激小鼠血清对有丝分裂原诱导的淋巴细胞增殖抑制的机理之一。     

活体染料CFDA-SE在淋巴细胞增殖研究中的应用

目的 :探讨活体染料CFDA SE在淋巴细胞增殖研究中的应用价值。方法 :利用CFDA SE染色、荧光抗体标记和流式细胞术 ,检测淋巴细胞及其亚群在多克隆刺激剂作用下荧光强度的变化 ,并应用相关软件分析其增殖情况。结果 :PDB ion omycin或ConA刺激 4 8h后均出现淋巴细胞分裂 ,表现为CFSE荧光强度的系列减半。环孢菌素A(CsA)可抑制ConA促进淋巴细胞增殖的作用 ,CFSE荧光无减半。ConA刺激4 8h后 ,出现CD4 T细胞和CD8 T细胞增殖不同步的现象 ,72h后这一现象更加明显。经ModFitTM 软件拟合后 ,得到的各项增殖指标显示 ,ConA对CD8 T细胞的促增殖作用强于对CD4 T细胞的作用。结论 :CFDA SE染色结合荧光抗体标记和流式细胞术 ,是分析淋巴细胞增殖的有力工具     

Studying the Proliferation of Human Peripheral Blood T Lymphocytes in Serum-Free Medium

We compared the cultivation of human peripheral blood lymphocytes in serum-free medium Hybris-2 and RPMI 1640 medium with 10% fetal bovine serum in the presence of phytohemagglutinin and interleukin-2. The optimal concentration of phytohemagglutinin significantly differed in serum-free and serum-containing media (0.5 and 5 mg/ml, respectively). Both mitogens were more potent in stimulating the proliferation of lymphocytes in serum-free medium than in serum-containing medium. Strong proliferation of CD3⁺ and CD4⁺ T lymphocytes was observed in both media. The dynamics of other markers was similar in serum-free and serum-containing media. However, significant differences were revealed between individual donors. Our results indicate that the developed serum-free medium may be used in lymphocyte cultivation for scientific, diagnostic, and therapeutic purposes. Translated from Kletochnye Tekhnologii v Biologii i Meditsine, No. 1, pp. 10-15, 2009 An erratum to this article can be found at     

Modulation of Mitogen-Induced Proliferation of Autologous Peripheral Blood Lymphocytes by Human Alveolar Macrophages

Experiments were carried out to determine the effect of cocultivation of T-cell-enriched human peripheral blood lymphocytes with autologous alveolar macrophages on mitogen-induced proliferation as determined by [³H]thymidine uptake. Cells obtained by fiberoptic bronchoscopy and saline bronchial lavage from 14 normal volunteers were enriched for macrophages by adherence in plastic dishes for 1 h in RPMI 1640 medium supplemented with 10% fetal calf serum. Nonadherent mononuclear cells were prepared from heparinized venous blood after Ficoll-Hypaque sedimentation by passage over nylon wool columns. T-cell-enriched populations were incubated with and without alveolar macrophages, either in the presence or absence of phytohemagglutinin. In these experiments, the number of lymphocytes was held constant (10⁵ per well), while the number of alveolar macrophages was varied (0.1 × 10⁵ to 4.0 × 10⁵ per well). Alveolar macrophages generally tended to stimulate phytohemagglutinin-induced lymphoproliferation at lymphocyte/macrophage ratios of 10:1 but consistently and significantly suppressed proliferation at ratios which approach those usually observed in recovered human bronchial lavage fluid, namely, 1:4. The suppressive effect of alveolar macrophages was observed as early as 48 h after culture initiation, while the magnitude of suppression increased with time. Suppression did not appear to be due to alteration in lymphocyte viability, nor was it sensitive to indomethacin. These results indicate that human alveolar macrophages can modulate the in vitro proliferative response of autologous peripheral blood lymphocytes. This observation may have relevance to interactions between alveolar macrophages and bronchial lymphocytes in the human lung in vivo.     

Nigerooligosaccharides Augments Mitogen-Induced Proliferation and Suppresses Activation-Induced Apoptosis of Human Peripheral Blood Mononuclear Cells

Nigerooligosaccharides (NOS), a mixture of nigerose and nigerosylmaltooligosaccharides, consists of immunopotentiating oligosaccharides found in foodstuffs. We have previously reported that activation of peripheral blood mononuclear cells (PBMC) in response to concanavalin A (Con A) or a streptococcal preparation of OK-432 is augmented in healthy young adults and elderly subjects after the intake of NOS-supplemented syrup. A reappraisal of the data suggests that NOS augments proliferation but partly suppresses activation-induced apoptosis of PBMC in response to these mitogens. To confirm this hypothesis, PBMC from healthy male subjects were stimulated with Con A or OK-432 in the presence of nigerose at the concentrations at which it was detected in the blood of subjects who had ingested NOS-supplemented syrup. Cellular activation, specifically metabolic demand, viability and proliferation, was assessed from glucose consumption, by WST-1 colorimetry and by 5-bromo-2'-deoxy-uridine incorporation assay, respectively. The Con A-induced activation of PBMC in each measurement was significantly augmented by nigerose. OK-432-induced decreases in the viability of PBMC were significantly inhibited by nigerose. Stimulation of PBMC with Con A or OK-432 induced apoptosis, but nigerose suppressed such activation-induced cell death. These results indicated that nigerose activated PBMC in vitro in a manner similar to the process observed in vivo, providing further evidence for the effectiveness of consumption of NOS-supplemented syrup.     

Triazolobenzodiazepines Exert Immunopotentiating Activities on Normal Human Peripheral Blood Lymphocytes

Previous studies have demonstrated that benzodiazepines (BDZ) (e.g. diazepam) inhibit immune responsiveness. Since these drugs are largely used in psychiatric patients it is of great importance to verify the existence of different types of BDZ, which are not suppressive for the immune system. In this framework, our results indicate that alprazolam and triazolam, two triazolo-BDZ, do not modify in vitro phagocytosis and killing exerted by normal human polimorphonuclear cells and monocytes. On the contrary, they significantly enhance T lymphocyte-dependent antibacterial activity in normal donors. These data support the concept that triazolo-BDZ and, in particular, alprazolam may represent more appropriate drugs for the treatment of psychiatric patients (e.g. patients with phobic disorders and/or migraine) who display immunodeficits.     

Triazolobenzodiazepines Exert Immunopotentiating Activities on Normal Human Peripheral Blood Lymphocytes

Abstract

Previous studies have demonstrated that benzodiazepines (BDZ) (e.g. diazepam) inhibit immune responsiveness. Since these drugs are largely used in psychiatric patients it is of great importance to verify the existence of different types of BDZ, which are not suppressive for the immune system. In this framework, our results indicate that alprazolam and triazolam, two triazolo-BDZ, do not modify in vitro phagocytosis and killing exerted by normal human polimorphonuclear cells and monocytes. On the contrary, they significantly enhance T lymphocyte-dependent antibacterial activity in normal donors. These data support the concept that triazolo-BDZ and, in particular, alprazolam may represent more appropriate drugs for the treatment of psychiatric patients (e.g. patients with phobic disorders and/or migraine) who display immunodeficits.     

Carbohydrate Metabolism in PHA Stimulated Human Peripheral Blood Lymphocytes

Human peripheral blood lymphocytes (PBL) stimulated in vitro by phytohemoagglutinin (PHA) manifest augmented glycolysis and oxidation of glucose-1-¹⁴C, indicating an increased utilization of the pentose pathway. Lactic acid production, as index of increased glycolysis, follows the same kinetic of thymidine incorporation and can he easily quantitated by an enzymatic assay.     

Ag85B表位多肽对体外单核细胞增殖影响的研究

目的通过观察Ag85B不同表位多肽对人外周血单个核细胞（PBMC）增殖的影响,筛选出增殖活性较强的Ag85B表位多肽,为制备结核新疫苗提供理论基础。方法采用的实验方法为人工预测Ag85B的表位,并合成多肽。将不同表位多肽体外刺激人外周血单个核细胞,利用CCK-8法测定单核细胞同一时相的吸光值,计算各自的增殖指数,并与BCG、Ag85B抗原等作对比。结果①Ag85B表位多肽组的OD450值及增殖指数均高于阴性对照组,且高于传统抗原BCG;②5条多肽中,28aa多肽的OD450值及增殖指数最高,且显著高于BCG（POD450=0.001〈0.01,P增殖指数=0.013〈0.05）,但低于PHA;③28aa多肽的OD450值及增殖指数明显低于Ag85B,差异有统计学意义（POD450=P增殖指数=0.000〈0.01）。结论预测的Ag85B表位多肽抗原具有一定的免疫原性,其中28aa多肽可作为优势表位多肽,但是否可用于表位多肽疫苗候选抗原还需进一步研究。     

Alpha-Fetoprotein on Human Peripheral Blood Lymphocytes Does Not Block Complement-dependent Lymphocytotoxicity

Using a direct immunofluorescence assay, we showed that alpha-fetoprotein (AFP) both in purified form and in hepatocellular carcinoma (HCC) sera was capable of binding onto 10-20% of T lymphocytes and 5-10% of B lymphocytes in human peripheral blood when these lymphocytes were preincubated in AFP-positive fluids at 4 degrees C in the presence of sodium azide. But when the preincubation temperature was raised to 37 degrees C, most of the membrane-bound AFP was internalized or shed, and, consequently, less than 3% of the cells showed positive membrane fluorescence. In addition, binding of AFP onto lymphocyte surface membrane and the continuous presence of large amounts of AFP in these lymphocyte cultures did not interfere with the action of cytotoxic antibodies directed against HLA determinants on the lymphocyte surface.     

Reactivity of Antiserum to Human Brain with Peripheral Blood Lymphocytes

Antisera to human brain (AHBS) and human thymocytes (AHTS) were produced in rabbits and selectively absorbed to render them specific for T cells. After absorption AHBS, but not AHTS, lost most of its cytotoxic activity against T cells. Absorbed AHBS bound up to 95% of peripheral blood T lymphocytes as detected by indirect immunofluorescence and inhibited up to 46% of the lytic activity of AHTS; however, it was incapable of inhibiting the E-rosette formation of T lymphocytes. All 10 samples of human peripheral blood lymphocytes, pretreated with AHBS, were significantly suppressed in their response to antigens, but fewer samples were affected in their response to mitogens and to allogeneic stimulation, indicating diversity in the nature of the receptors involved in the cellular responses.