鼻咽癌病人血浆脂质过氧化物,IL—2及IL—2受体活性的研究

本文对不同病期鼻咽癌(NPC)病人血浆中脂质过氧化物—丙二醛(MDA)、外周血淋巴细胞产生IL-2的能力、IL-2受体活性进行初步探讨。结果显示NPC病人血浆中MDA含量较正常人明显增高(P<0.01),晚期病人比早期病人增高更明显(P<0.01);患者外周血淋巴细胞分泌IL-2的能力及IL-2受体活性较正常人明显下降(P<0.01)。这提示了NPC患者血浆中MDA的增高和肿瘤的发生发展有关,NPC患者免疫功能的下降可能与MDA的升高有关。     

IgA肾病患者IL—2的产生及IL—2受体的表达

本文对IgA肾病(IgAN)外周血淋巴细胞白细胞介素2(IL-2)的产生、受体的表达及免疫球蛋白的产生进行了研究。结果发现:外周血淋巴细胞产生IL-2的活性明显增高,IL-2受体表达亦明显增强并伴有免疫球蛋白产生增多,提示IgAN存在着细胞免疫功能的紊乱。     

黄芪对反复呼吸道感染患者体内SIL-2R、IL-8及Ig的影响

目的:探讨黄芪治疗反复呼吸道感染(RRI)过程患者体内白介素-2 受体(SIL-2R)、白介素-6(IL-6) 和免疫球蛋白(Ig) 的变化.方法:将60例RRI患者随机分为2组,2组均给予抗感染及对症治疗,治疗组在此基础上加用黄芪注射液治疗,采用双抗体夹心ELISA 法测定60例RRI患者血清SIL-2R 和IL-6;速率散射比浊法测定Ig.结果:RRI患者SIL-2R,IL-8水平明显高于正常组(P<0.01),Ig 水平低于正常组,治疗后2组SIL-2R、IL-8均下降(P<0.05),两组间比较有统计学意义(P<0.05),治疗前后,治疗组Ig水平明显上升,对照组则无明显变化,两组比较有统计学意义(P<0.01).结论:SIL-2R、IL-8及Ig参与了RRI 的免疫发病机制,黄芪可提高RRI患者的免疫力.     

血清IL-2、TNF-α及IL-13在类风湿关节炎诊断与治疗中的临床意义

为探讨血清IL-2、TNF-α及IL-13在类风湿关节炎(rheumatoid arthritis,RA)诊断与治疗中的临床意义,收集了30例RA患者外周血,其中经过常规治疗的患者27例,常规治疗合并生物制剂治疗的患者3例,以30例健康体检者为正常对照,采用酶联免疫吸附试验双抗体夹心法检测RA患者治疗前后血清中IL-2、TNF-α及IL-13的表达水平,并分析IL-2、TNF-α及IL-13与临床指标RF、CRP、ESR的相关性。结果发现RA患者治疗前血清IL-2、TNF-α及IL-13表达水平明显高于健康对照组(P<0.05),常规治疗后IL-2表达水平降低(P<0.05),常规治疗合并生物制剂治疗后IL-2、IL-13表达水平明显降低(P<0.05);RA患者发病期血清IL-2、TNF-α及IL-13与RF、CRP、ESR呈正相关(P<0.05)。该研究提示RA患者血清中IL-2、TNF-α及IL-13的表达在RA的诊断与治疗中有一定的临床意义。     

类风湿关节炎患者血清和关节液细胞因子水平的变化及临床意义

目的探讨幼年类风湿关节炎(JRA)及类风湿关节炎(RA)患者IL-6、IL-8、sIL-2R和TNF-α等细胞因子(CK)水平的变化,及其与风湿活动的传统指标血沉(ESR)和C-反应蛋白(CRP)的相关性。方法采用夹心ELISA法,对30例JRA和34例RA患者的血清中,4例JRA、7例RA、6例骨性关节炎(OA)和9例半月板损伤(MT)患者的关节液中IL-6、IL-8、sIL-2R和TNF-α的水平进行检测。结果①30例JRA、34例RA患者血清IL-6和sIL-2R的水平与对照组相差非常显著(P〈0.01);30例JRA患者血清IL-8水平与对照组比较相差显著(P〈0.05)。②JRA全身型、少关节型患者血清IL-8、sIL-2R的水平和JRA多关节型患者血清IL-6的水平与对照组相差非常显著(P〈0.01)。③4例JRA及7例RA患者关节液sIL-2R的水平和RA患者关节液的IL-6水平与对照组相差显著(P〈0.05)。④JRA患者血清IL-6和sIL-2R的水平与ESR和CRP的变化呈明显的相关关系(r值分别为0.532和0.621)。结论①IL-6、sIL-2R的水平与JRA、RA病的活动性有关,是类风湿活动性的主要指标。②sIL-2R不仅参与JRA和RA的全身病理损伤,而且是引起关节局部损伤的主要CK,IL-6也参与JRA关节局部的病理损伤,在RA关节局部损伤似乎更为重要。③IL-8主要参与JRA的全身病理损伤,对关节局部病理损伤似乎并不重要。     

类风湿关节炎T细胞亚群4-1BB的表达与其分泌TH1/TH2细胞因子的关系

目的探讨体外培养的类风湿关节炎(rheumatoid arthritis,RA)T淋巴细胞上共刺激分子4-1BB的表达,及与其分泌T_H1/T_H2细胞因子的关系。方法应用流式细胞术检测体外培养的30例RA患者和20例正常对照者T细胞活化前后4-1BB的表达,并且应用ELISA方法检测培养上清液中IFN-γ、IL-4水平。结果RA患者CD4~ T和CD8~ T细胞表达的4-1BB明显高于正常对照组(表达百分率分别为18.56%±4.08%和10.33%±2.13%;1.24%±0.12%和0.87%±0.09%,P<0.01),经抗CD3单抗体外刺激后CD4~ T和CD8~ T细胞表达的4-1BB均显著高于活化前(表达百分率为33.21%±4.29%和21.35%±8.12%,P<0.01)。RA患者CD4~ T/CD8~ T比值明显升高,而且与4- 1BB~ CD4~ T细胞数呈正相关关系(r=0.84,P<0.01),RA患者T淋巴细胞培养液上清中IFN-γ、IL-4均高于正常对照组(P<0.05),经抗CD3单抗体外刺激后上清液中IFN-γ及Ib-4均明显高于活化前,以IFN-γ升高最为显著(P<0.01)。而且抗CD3单抗刺激前后4-1BB~ CD4~ T细胞数与培养上清液中IFN-γ水平均呈正相关关系(r=0.721,r=0.487,P<0.05)。结论类风湿关节炎患者T细胞表达的4-1BB在类风湿关节炎的发生发展中具有重要意义,4-1BB可能通过对CD4~ T细胞活化,促使T_H1细胞因子分泌,参与关节炎症和免疫损伤。     

儿童肺结核病患者血清IL-2、SIL-2R和VEGF水平及临床意义

目的:探讨了儿童肺结核病患者血清白细胞介素-2(IL-2)、可溶性白细胞介素-2受体(SIL-2R)和血管内皮生长因子(VEGF)水平的变化及意义。方法:分别应用放射免疫分析和ELISA对68例儿童肺结核病患者进行了血清IL-2、SIL-2R和VEGF水平观察,并与35名正常健康人作比较。结果:活动性儿童肺结核病患者血清IL-2水平显著低于正常儿组(P<0.01),而SIL-2R和VEGF水平显著高于正常儿组(P<0.01)。结论:测定儿童肺结核病患者血清IL-2、SIL-2R和VEGF水平更有助于儿童肺结核病活动性的检测,对该病的预防和预后均有一定的意义。     

人外周血淋巴细胞IL-2R_α表达特性及动力学

本文研究了PHA诱导人外周血淋巴细胞(PBl)IL-2Rα表达的动力学,并探讨了TPA、5-氮胞苷、羟基脲、放线菌素D和秋水仙素等对PHA作用的影响。新分离的PBL几乎不表达IL-2Rα。接受PHA刺激12h时,IL-2Rα表达即明显增加,72h达高峰,随后逐渐降下,至第七天接近未刺激水平。TPA、5-氮胞苷可增加PHA的作用,羟基脲和秋水仙素不影响,而放线菌素D可抑制pHA的效应。这表明人PBL IL-2Rα表达在转录水平调节,与DNA合成及细胞有丝分裂关系不大。体外培养的淋巴母细胞再次接触上述试剂时与初次接触呈现相同的反应性。     

类风湿性关节炎及骨关节炎患者外周血单个核细胞及滑膜成纤维细胞中白细胞介素27水平增高

目的检测类风湿性关节炎(RA)及骨关节炎(OA)外周血单个核细胞(PBMC)和滑膜成纤维细胞(FLS)白细胞介素27(IL-27)及其受体的水平。方法收集20例RA、20例OA、20例正常人PBMC和4例RA及4例OA患者滑膜组织,培养FLS,采用实时荧光定量PCR法检测PBMC和FLS中IL-27 mRNA的水平;采用免疫细胞化学法检测RA、OA患者FLS IL-27Rα的表达。结果 RA及OA患者PBMC中IL-27 mRNA水平分别是正常人的1.81倍及2.07倍;RA患者和OA患者PBMC中IL-27mRNA水平无明显差异,但RA患者FLS中IL-27 mRNA是OA患者的3.74倍;RA组FLS的IL-27Rα较OA组水平增高。结论 RA、OA患者IL-27水平增加,且RA患者FLS中IL-27水平高于PBMC。     

TYK2 基因在类风湿关节炎中的表达及作用研究

目的:观察酪氨酸激酶2(TYK2)基因在类风湿关节炎(RA)患者中的表达,并探讨其与RA标志物、炎症因子及骨代谢指标的相关性。方法:选取RA患者55例、健康志愿者50例检测其血清类风湿因子(RF)、抗环瓜氨酸肽抗体(抗CCP)、IL-2、IL-17、IL-21、骨钙素(OST)、25羟基维生素D[25(OH)D]、β-胶原特殊序列(β-CTX)、总1型胶原氨基端延长肽(T-P1NP)水平,RT-PCR法检测TYK2 mRNA表达,并对其进行相关性分析。结果:与健康志愿者相比,RA患者外周血RF和抗CCP水平显著升高(P0.01),血清IL-2、IL-17和IL-21水平明显升高(P0.01),骨代谢指标OST、25(OH)D、T-P1NP水平明显降低,β-CTX水平明显升高(P0.01),外周血单核细胞TYK2 mRNA表达明显升高。TYK2 mRNA与RF、抗CCP、IL-2、IL-17、IL-21、β-CTX呈显著正相关,与OST、25(OH)D、T-P1NP呈显著负相关。结论:TYK2基因的表达与RA病情进展密切相关,可能通过调控与其相关的IL-2、IL-17和IL-21水平参与RA发展。     

A Comparative Study of Interleukin-1β Production and P2x7 Expression After Atp Stimulation by Peripheral Blood Mononuclear Cells Isolated From Rheumatoid Arthritis Patients and Normal Healthy Controls

Interleukin 1 beta (IL-1β) is a proinflammatory cytokine that is considered to play an important role in the progression of rheumatoid arthritis (RA). A stimulus such as ATP is necessary to cause the release of mature IL-1β, via activation of the P2X₇ receptor on monocytes. In this study, the production of IL-1β in whole blood after ATP stimulation and expression of P2X₇ receptors in RA and healthy subjects were examined. Blood samples from RA patients or healthy controls were stimulated with ATP in the presence of lipopolysaccharide (LPS). Supernatants were harvested and IL-1β levels were measured by enzyme-linked immunosorbent assay (ELISA). Expression of P2X₇ receptors was measured using flow cytometry. ATP induced significantly higher levels of IL-1β in LPS-activated RA blood samples compared to controls. A significant up-regulation of P2X₇ receptor expression on mononuclear cells was observed after overnight incubation with ATP without any significant differences between RA patients and normals. These data suggest that RA patient mononuclear cells are more sensitive to ATP stimulation than healthy individuals perhaps due to genetic polymorphism in the P2X₇ gene.     

The prostaglandin E2 increases the production of IL-17 and the expression of costimulatory molecules on γδ T cells in rheumatoid arthritis

γδ T cells play important roles in the development of rheumatoid arthritis (RA) through their antigen-presenting capacity, release of pro-inflammatory cytokines, immunomodulatory properties, interaction with CD4⁺CD25⁺ Tregs and promotion of antibody production by helping B cells. Although prostaglandin E2 (PGE2) was proved to have the ability to enhance the antigen-presenting function of dendritic cells and IL-17 production of CD4⁺ αβ T cells in RA, the role of PGE2 in γδ T cells from RA disease has not yet been clarified. The goal of this study was to determine the role of PGE2 in γδ T cells in RA. We first demonstrated that the population of γδT17 cells increased in patients with RA compared to healthy controls. Then, IL-17A level in patients with RA was shown to increase compared to healthy controls. After adding PGE2 to γδ T cells from patients with RA, the IL-17A level increased accordingly, and the expression of the costimulatory molecules, CD80 and CD86, on these cells also increased. These results suggest that PEG2 can increase the production of IL-17A and the expression of CD80 and CD86 on γδ T cells in patients with RA. These findings will benefit to explore new therapeutic targets for RA disease.     

Peripheral blood mononuclear cells from patients with rheumatoid arthritis spontaneously secrete vascular endothelial growth factor (VEGF): specific up-regulation by tumour necrosis factor-alpha (TNF-alpha) in synovial fluid.

This study was designed to investigate VEGF production from peripheral blood mononuclear cells (PBMC) from patients with rheumatoid arthritis (RA) compared with healthy controls and to identify the predominant cellular source in PBMC isolated from RA patients. The regulation of PBMC VEGF production by cytokines and synovial fluid (SF) was studied. PBMC were isolated from RA patients and healthy controls and stimulated with lipopolysaccharide (LPS), IL-1beta, IL-4, IL-6, IL-8, IL-10, TNF-alpha and transforming growth factor-beta (TGF-beta) isoforms for varying time points up to 72 h at 37 degrees C/5% CO2. The effect of SF on VEGF secretion by PBMC was also studied. Supernatant VEGF levels were measured using a flt-1 receptor capture ELISA. RA patients had significantly higher spontaneous production of VEGF compared with controls, and monocytes were identified as the predominant cellular source. RA PBMC VEGF production was up-regulated by TGF-beta isoforms and TNF-alpha and down-regulated by IL-4 and IL-10, with no effect observed with IL-1beta, IL-6 and IL-8. Antibody blocking experiments confirmed that TNF-alpha and not TGF-beta isoforms in SF increased VEGF secretion by RA PBMC. These results emphasize the importance of monocytes as a source of VEGF in the pathophysiology of RA. Several cytokines known to be present in SF can modulate the level of VEGF secretion, but the predominant effect of SF in VEGF up-regulation is shown to be dependent on TNF-alpha.     

Modulation of human T cell functions by surface sulphydryl groups: differential effects on IL-2 production and responsiveness.

An impermeable thiol blocker has been used to investigate the role of sulphydryl (SH) groups in the production of and responsiveness to IL-2 by normal human T lymphocytes. Surface SH blockade of mononuclear cells prior to incubation with mitogen (phytohaemagglutinin, concanavalin A, CD3 MoAb) had no effect on production of IL-2 but markedly impaired cellular responsiveness to exogenous IL-2. Studies using MoAbs indicated that this effect was accompanied by decreased expression of both the CD25 and p75 subunits of the IL-2 receptor. Blocking surface SH groups did not affect binding of IL-2 to p75 on unstimulated mononuclear cells, but inhibited binding to high-affinity receptors on a T lymphoma cell line. The data are consistent with the hypothesis that sulphydryl groups on the IL-2 receptor are required for its function and may be involved in the interaction of the CD25 and p75 subunits leading to generation of the high-affinity binding site. The surface thiol identified on the IL-2 receptor may be a candidate for oxidation on cells from patients with chronic inflammatory diseases such as rheumatoid arthritis and thus contribute to the aberrant function of T cells in these patients.     

Monocytes from patients with rheumatoid arthritis and type 2 diabetes mellitus display an increased production of interleukin (IL)-1β via the nucleotide-binding domain and leucine-rich repeat containing family pyrin 3(NLRP3)-inflammasome activation: a possible implication for therapeutic decision in these patients

A better understanding about the mechanisms involved in the pathogenesis of type 2 diabetes mellitus (T2D) showed that inflammatory cytokines such as tumour necrosis factor (TNF) and interleukin (IL)-1β play a pivotal role, mirroring data largely reported in rheumatoid arthritis (RA). IL-1β is produced mainly by monocytes (MO), and hyperglycaemia may be able to modulate, in the cytoplasm of these cells, the assembly of a nucleotide-binding domain and leucine-rich repeat containing family pyrin (NLRP3)-inflammosome, a cytosolic multi-protein platform where the inactive pro-IL-1β is cleaved into active form, via caspase-1 activity. In this paper, we evaluated the production of IL-1 β and TNF, in peripheral blood MO of patients affected by RA or T2D or both diseases, in order to understand if an alteration of the glucose metabolism may influence their proinflammatory status. Our data showed, after 24 h of incubation with different glucose concentrations, a significantly increased production of IL-1β and TNF in all evaluated groups when compared with healthy controls. However, a significant increase of IL-1β secretion by T2D/RA was observed when compared with other groups. The analysis of relative mRNA expression confirmed these data. After 24 h of incubation with different concentrations of glucose, our results showed a significant increase in NLRP3 expression. In this work, an increased production of IL-1β by MO obtained from patients affected by both RA and T2D via NLRP3-inflammasome activation may suggest a potential IL-1β targeted therapy in these patients.     

IL-2 regulation of soluble IL-2 receptor levels following thermal injury.

In the immunosuppressed burn patient serum levels of both IL-2 and a soluble form of IL-2 receptor alpha (sIL-2R alpha) are significantly elevated. Strikingly, the production of these markers by the in vitro activated patients' cells is decreased. This study examines the role of IL-2 in the decreased production of the sIL-2R alpha in vitro in patients with major burns (n = 18, 30 to greater than 70% total body surface area). Peripheral blood mononuclear cell (PBMC) cultures from patients with highly elevated serum sIL-2R alpha, and from healthy controls (n = 12) were activated with concanavalin A (Con A) at initiation. In patients' cultures mitogen-induced increments of sIL-2R alpha levels were significantly lower. There was a significant negative correlation (r = 0.64, P less than 0.001) between a high serum sIL-2R alpha level and a decreased lectin-induced sIL-2R alpha release in vitro. Low levels of sIL-2R alpha in patients' samples were not normalized by increasing the number of T lymphocytes. Also exogenous rIL-1 was without effect, whereas rIL-3 increased sIL-2R alpha release in some cultures. However, sIL-2R alpha levels were significantly increased in patients' cultures by (i) addition of exogenous IL-2; (ii) removal of adherent cells; (iii) addition of cyclooxygenase inhibitor, indomethacin; (iv) bypassing cell surface activation by the combination of the calcium ionophore A23187 and the phorbol ester 12-o-tetradecanoyl acetate. The cyclic AMP-elevating drug, forskolin, abrogated the ability of exogenous IL-2 to increase sIL-2R alpha production. Thus, in the burn patient, the reduced in vitro sIL-2R alpha release appears to relate to abnormalities in IL-2 production and action mediated through its functional surface receptor. Elevated levels of sIL-2R alpha in vivo may, therefore, reflect systemic activation of T lymphocytes in response to biologically active IL-2.     

Th1 (IL-2, interferon-gamma (IFN-γ)) and Th2 (IL-10, IL-4) cytokine production by peripheral blood mononuclear cells (PBMC) from patients with systemic lupus erythematosus (SLE)

We investigated the production of IL-2, IFN-γ, IL-10 and IL-4 by PBMC from 24 patients with SLE and 10 healthy individuals. Basal and mitogen-stimulated (lipopolysaccharide and phytohaemagglutinin (LPS + PHA)) cytokine production was determined in a whole blood assay (WBA). Supernatants were collected and assayed with specific ELISAs. Although the IL-2 and IFN-γ contents did not differ significantly between patients and controls under both conditions, statistically significant correlations were found between each cytokine and disease activity (SLAM index) after stimulation (respectively, r= 0.501, P = 0.01 and r = 0.631, P = 0.001). PBMC IL-10 production was significantly higher for patients than controls (P = 0.05), but no correlation between IL-10 levels and the SLAM index was obtained. IL-4 production was not statistically different between SLE patients and controls. For stimulated WBAs, the IL-10/IL-2 and IL-10/IFN-γ ratios were significantly correlated with disease severity (P = 0.02; P = 0.001, respectively). Overall, our data suggest that SLE is characterized by an elevated production of IL-10, reflecting the basal state of activation of the immune system. During exacerbation of SLE, IL-2 and IFN-γ are synthesized in larger amounts and may cause the tissue damage observed.