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。     

类风湿关节炎湿热痹阻型患者血清和关节液IL-1、IL-6、TNF-α的表达研究

目的检测类风湿关节炎(RA)湿热痹阻型患者血清和关节液中IL-1、IL-6、TNF-α的表达水平,分析IL-1、IL-6、TNF-α与炎性活动指标ESR、CRP、DAS28之间的关系,探索细胞因子与RA发生发展的关系。方法采用ELISA法检测RA湿热痹阻型患者外周血血清90例、RA湿热痹阻型患者关节液28例、骨性关节炎患者关节液30例及健康对照组血清30例IL-1、IL-6、TNF-α的表达,并常规方法检测ESR、CRP。利用SPSS11.5统计学软件进行统计学分析。结果 RA湿热痹阻型患者外周血血清中IL-1、IL-6及TNF-α表达水平显著高于健康组血清,差异有统计学意义(t=12.25,10.56,8.758;P0.05);RA湿热痹阻型患者关节液中IL-1、IL-6及TNF-α的表达水平显著高于外周血血清,差异有统计学意义(t=4.35,3.09,2.496;P0.05);RA湿热痹阻型患者关节液中IL-1、IL-6及TNF-α表达水平明显高于骨性关节炎关节液,差异有统计学意义(t=3.47,2.46,2.51;P0.05)。RA湿热痹阻型患者外周血血清和关节液中IL-1、IL-6及TNF-α的表达与ESR、CRP、DAS28无相关性(P0.05)。结论 RA湿热痹阻型患者外周血和关节液中IL-1、IL-6及TNF-α的异常升高可能参与了RA的发生发展。     

类风湿性关节炎患者IL-6、IL-18和CRP的水平变化及意义

目的：研究类风湿性关节炎（RA）患者血清白细胞介素-6（IL-6）、白细胞介素-18（IL-18）和C-反应蛋白（CRP）水平的变化及其临床意义。方法：收集84例RA患者,以70例健康体检者作对照。采用双抗体夹心酶联免疫吸附法测定血清IL-6、IL-18和免疫荧光法测定CRP的水平,并测定血小板计数（Plt）、血沉（ESR）、类风湿因子（RF）。结果：RA患者的血清Plt、ESR、RF、IL-6、IL-18和CRP的含量明显高于健康对照组（P〈0.01）。RA患者活动期上述指标含量（除RF外）均显著高于稳定期（P〈0.01）,Plt升高RA患者组与Plt正常组相比,RF、ESR、IL-6、IL-18和CRP水平均有明显统计学差异（P〈0.05）。结论：IL-6、IL-18和CRP在RA患者的疾病发展过程中发挥着重要作用,它们的水平变化与RA患者病情有关,联合动态监测有助于临床观察RA患者的病情变化和治疗效果。     

类风湿关节炎患者单个核细胞IL-23的表达及意义

目的:通过分析比较类风湿关节炎(Rheumatoid Arthritis,RA)患者血清和外周血单个核细胞(Peripheral bloodmononuclear cell,PBMC)IL-23的表达,探讨其在RA发病过程中的作用,以期为治疗开辟新途径。方法:应用ELISA方法检测RA、骨性关节炎(Osteoarthritis,OA)及正常对照组血清中IL-23和TNFα-蛋白水平,采用RT-PCR法测定各组PBMC中IL-23p19mRNA的表达,并作血清IL-23含量、PBMC IL-23p19mRNA的表达与血清TNFα-水平的相关分析。结果:RA组血清IL-23水平高于正常对照组(P=0.001),其他组间无统计学差异(分别为P=0.122,P=0.127);而各组血清TNFα-的水平均具有统计学差异,RA和OA组均高于对照组(P=0.000,P=0.042),RA组也高于OA组(P=0.013);RA组IL-23p19mRNA的表达高于OA和正常对照组(P=0.000,P=0.000),而OA组与正常对照组差异无统计学意义(P=0.628);血清中IL-23和TNFα-的相关性无统计学差异(r=0.212,P=0.262),而PBMC中IL-23p19mRNA的表达与血清TNFα-呈正相关关系(r=0.392,P=0.032)。结论:IL-23在RA患者PBMC中高表达,在RA的炎性发展过程起到了重要作用。     

Th22 细胞及其功能分子IL-22 在狼疮性肾炎中的作用研究

目的:观察狼疮性肾炎(LN)患者外周血中的Th22 细胞及其功能分子白细胞介素(IL)-22 的表达,并探讨其意义。方法:选择系统性红斑狼疮患者(无肾脏受累)38 例,狼疮性肾炎患者32 例,健康志愿者10 例,分别作为SLE 组、LN 组和健康对照组(HC 组)。应用流式细胞仪检测其外周血中Th22 细胞,酶联免疫吸附实验检测患者血清中IL-22 的水平。比较三组Th22 细胞和血清中IL-22 水平差异,分析LN 组Th22 细胞和IL-22 与SLE 疾病活动指数评分(SLEDAI)的相关性。结果:SLE 组和LN 组患者外周血中Th22 细胞较健康对照组明显升高(P<0.01),血清中IL-22 水平也较健康对照组明显升高(P<0.01);Th22 和IL-22 在LN 组较SLE 组要稍微高表达,但差异均无统计学意义(P>0.05);LN 组患者Th22 细胞比例和IL-22水平与对应患者的疾病活动性评分SLEDAI 呈正相关。结论:Th22 细胞和其功能分子IL-22 参与SLE 和LN 的发病和进展。     

正五聚体蛋白3、IL-6和CRP水平与RA疾病活动度的相关性

为探讨正五聚体蛋白3(pentraxin 3, PTX-3)、IL-6和CRP水平与RA疾病活动度的相关性,以90例RA患者为研究组,其中疾病低度活动者30例(33.3%),中度活动者44例(48.9%),高度活动者16例(17.8%)。同期纳入60例健康者为对照组,分析研究组与对照组血清中PTX-3、IL-6及CRP水平的差异。结果显示,研究组中PTX-3和CRP水平远高于对照组(P0.001)。随着RA病情的发展,疾病活动指数DAS 28评分越高的患者,IL-6和CRP水平越高(均P0.001),同时,ESR在高度活动组中水平更高(P0.001)。此外,低、中、高度活动组中PTX-3水平均远高于对照组(均P0.001)。CRP、IL-6和ESR水平与DAS 28评分呈低度正相关(r=0.654,P0.001;r=0.555,P0.001;r=0.557,P0.001),IL-6与ESR水平也呈正相关(r=0.489,P0.001)。由此PTX-3、IL-6、CRP可作为RA的生物标志物,几项指标的联合监测可有效反映病情发展状况,为临床治疗提供实验依据。     

IL-23及其受体与类风湿关节炎疾病活动的相关性分析

为探讨血清IL-23水平与类风湿关节炎(rheumatoid arthritis,RA)病情活动相关性及对患者外周血NK细胞功能的影响,选取本院2015年2月至2016年4月收治的RA患者80例作为观察组,选择健康者80例作为对照组,对两组入选者行血清IL-23、外周血NK细胞及相关指标检测,回顾性分析检测结果显示观察组患者IL-23显著高于对照组(t=87.217,P0.05);观察组IL-23R在NK细胞的表达率为(1.23±0.28)%,显著低于对照组的(4.53±1.07)%(t=21.097,P0.05);稳定与轻度、中度、重度活动组以及X线分期Ⅰ、Ⅱ、Ⅲ、Ⅳ期组患者IL-23水平均呈递增趋势,且两两对比差异显著(P0.05);观察组患者IL-23水平与RF、ESR、CCP、CRP、DAS28评分均表现为显著正相关(P0.05),由此IL-23水平与RA病情进展密切相关。     

类风湿关节炎患者血清中IL-17 表达及IL-17F 基因位点rs763780 多态性分析

目的:了解类风湿关节炎(RA)患者血清中IL-17、类风湿因子(RF)及C反应蛋白(CRP)的表达,并探讨IL-17F基因位点rs763780多态性与RA发病易感相关性。方法:收集2016年1月～2018年10月来深圳市福田区风湿病专科医院就诊的RA患者176例为研究组,并选取同期来院体检的健康人群110名为对照组,分别检测血清中IL-17、RF及CRP含量,并采用聚合酶链反应(PCR-RFLP)技术对IL-17F基因位点rs763780多态性进行分析。结果:RA组血清中IL-17、RF及CRP含量分别为(36.73±11.06)pg/ml,(58.05±14.92)U/ml和(32.15±13.96)mg/L,明显高于对照组的(14.52±6.35)pg/ml,(10.24±5.32)U/ml和(6.81±3.25)mg/L,差异均有统计学意义(t=3.035 7～5.320 6,P0.05);RA组CC基因型和C等位基因检出率分别为19.32%和41.48%,明显高于对照组的6.36%和26.82%,差异有统计学意义(χ~2=3.916 5～5.054 7,P0.05);RA组IL-17F基因位点rs763780 CC基因型中的IL-17水平为(42.96±13.64)pg/ml,明显高于其他基因型,差异有统计学意义(P0.05),而三种基因型血清中RF和CRP水平之间差异均无统计学意义(P0.05)。结论:RA患者血清中IL-17,RF及CRP水平明显高于健康人群,同时RA患者IL-17F基因位点rs763780 CC基因型及C等位基因检测率明显高于健康人群,可能是导致本地区RA发病的危险遗传易感基因之一。     

IL-22在类风湿关节炎中的表达及临床意义

目的：探讨RA患者IL-22的表达水平并分析其临床意义。方法：流式细胞术检测RA患者（50例）、OA患者（15例）、健康对照组（15例）外周血及RA关节滑液Th22细胞所占比例,ELISA法检测血清及RA关节滑液中IL-22水平。检测RA患者疾病活动性、临床指标、骨破坏情况,分析其与IL-22水平相关性。两组间比较用t检验,相关性分析采用Pearson直线相关分析法,多组样本比较用Kruskal-Wallis H检验。结果：RA患者外周血中Th22细胞比例高于OA组（t=2.290,P=0.021）及健康对照组（t=2.524,P=0.015）;RA患者血清IL-22水平高于OA患者（t=2.560,P=0.014）及健康对照组（t=2.768,P=0.009）。RF阳性RA血清IL-22水平高于RF阴性RA（t=2.322,P=0.035）。抗CCP抗体阳性RA血清IL-22水平高于抗CCP抗体阴性RA（t=2.504,P=0.015）。RA血清IL-22水平与ESR、RF、DAS28评分呈正相关关系（r分别为0.312、0.314、0.332,P<0.05）。RA患者血清IL-22水平随骨侵蚀的加重呈递增趋势,X线不同分期之间差异有统计学意义（H=9.14, χ²_0.05（3）=7.81,H>χ²_0.05(3),P<0.05)。有关节积液RA患者血清IL-22水平高于无关节积液RA患者（t=2.587,P=0.012）,关节滑液IL-22水平高于血清（t=2.668,P=0.011）,与关节滑液Th22细胞比例无相关性（r=0.287,P=0.065）。结论：RA患者血清及关节滑液IL-22水平增高,与疾病活动性及骨破坏相关,可成为病情评估与骨破坏的指标。IL-22可能为RA治疗的新靶点。     

早期RA患者血清炎症指标与临床特征、转归的相关性研究

为探讨早期RA患者血清炎症指标与临床特征、转归的相关性,选择2010年6月至2015年6月于赣南医学院第三附属医院收治的72例RA患者作为RA组,另选择70例同期健康体检者为健康组。采用免疫比浊法检测血清CRP及RF,ELISA双抗体夹心法检测抗环瓜氨酸肽(cyclic citrullaminopeptide,CCP)抗体,魏氏法检测ESR。比较RA患者服药前后临床指标及转归情况,同时采用Pearson相关性分析法对RA患者血清炎症指标与临床特征、转归的关系进行分析。结果显示,RA组CRP、ESR、RF及抗CCP抗体水平均显著高于健康组(P0.05)。治疗后RA患者晨僵时间、休息痛、健康评估问卷等临床指标和CRP、RF、ESR等炎症指标均显著低于治疗前(P0.05)。所有患者治疗后病情均有不同程度好转,早期RA患者转归分为三类:43例缓慢逐步进展,20例进展迅速,9例可控制。DAS28积分与CRP、ESR、RF及抗CCP抗体的相关系数分别为0.266、0.255、0.291和0.382(P0.05);转归与CRP、ESR、RF及抗CCP抗体的相关系数分别为0.195、0.269、0.288和0.355(P0.05)。多元逐步回归分析结果显示,CRP、ESR、RF及抗CCP抗体与DAS28积分及转归呈正相关。提示联合检测CRP、ESR、RF及抗CCP抗体对于评估早期RA患者疾病活动度及转归具有一定价值。     

IL-10 subfamily members: IL-19, IL-20, IL-22, IL-24 and IL-26

It has been reported that the CD4+ T cell is a very important source of interleukin 10 (IL-10), while CD8+ cells produce low amounts. IL-10 exerts several immune stimulating, as well as inhibitory effects. There are at least five novel human IL-10 family-related molecules: IL-19, IL-20, IL-22, IL-24, and IL-26. Activated T cells produce IL-19, IL-22 and IL-26, while IL-24 is produced by activated monocytes and T-cells. IL-20 induces cheratin proliferation and Stat-3 signal transduction pathway, while IL-22 induces acute-phase production by hepatocytes and neonatal lethality with skin abnormalities reminiscent of psoriasic lesions in humans. In addition, IL-22 mediates inflammation and binds class II cytokine receptor heterodimers IL-22 RA1/CRF2-4. This cytokine is also involved in immuno-regulatory responses. IL-26 (AK155) is a novel cytokine generated by memory cells and is involved in the transformed phenotype of human T cells after infection by herpes virus. All these new IL-10 subfamily member cytokines are strongly involved in immune regulation and inflammatory responses.     

The extended IL-10 superfamily: IL-10, IL-19, IL-20, IL-22, IL-24, IL-26, IL-28, and IL-29

Cytokines are involved in virtually every aspect of immunity and inflammation. A cascade of responses evolves after cytokine activation, although optimal function might ultimately involve several complementary cytokines. Understanding the function of individual cytokines is complicated because their role can vary depending on the cellular source, target, and phase of the immune response. In fact, numerous cytokines have both proinflammatory and anti-inflammatory potential, with the contrasting outcome observed being determined by the immune cells present and their state of responsiveness to the cytokine. These issues make the study of cytokine biology daunting, particularly so for IL-10 and IL-10-related genes. The IL-10 superfamily is highly pleiotropic. These genes are linked together through genetic similarity and intron-exon gene structure. Significant commonality exists not only through shared receptors but also through conserved signaling cascades. However, its members mediate diverse activities, including immune suppression, enhanced antibacterial and antiviral immunity, antitumor activity, and promotion of self-tolerance in autoimmune diseases.     

IL-21 and IL-21 receptor

Interleukin (IL)-21 is a new member of the type I cytokine superfamily. Although it is most homologous to IL-15, it has a unique receptor chain, IL-21R, that pairs with the γ-common cytokine receptor chain. The first experiments examining the biology of the IL-21 pathway reveal that it is a cytokine with effects on natural killer (NK) cells, T cells, and B cells. Mice deficient in the IL-21 R have also been made, and are being examined for the effects of the IL-21/IL-21R pathway in vivo. Here we summarize our current knowledge of this new cytokine pathway, and its role in innate and adaptive immunity.     

IL-4、IL-13蛋白表达及抗IL-4、IL-13人源单链抗体的筛选

目的:表达IL-4和IL-13蛋白,从人源单链抗体文库中分别筛选抗IL-4和抗IL-13单链抗体.方法:采用RT-PCR从健康志愿者外周血单核细胞(PBMC) mRNA中扩增IL-4和IL-13 cDNA;构建硫氧还蛋白融合表达载体,转化大肠杆菌BL21,IPTG诱导表达并对表达产物进行纯化鉴定.以生物素化的IL-4和IL-13为抗原从前期构建的人源抗体文库中采用噬菌体展示技术分别筛选抗IL-4和抗IL-13人源单链抗体(scFv).结果:扩增的IL-4 cDNA大小为280 bp,表达的融合蛋白大小为27 kD左右.扩增的IL-13 cDNA大小为252 bp,表达的融合蛋白大小为25 kD左右.分别以生物素化的IL-4和IL-13蛋白为抗原,采用噬菌体展示技术对人源抗体文库进行3轮富集后,分别有大约37％的scFvs与IL-4有结合特性,有约27％的scFvs与IL-13有结合特性.筛选了4株分别与IL-4和IL-13结合能力强的单链抗体进行了Westem blot鉴定和测序.结论:成功筛选到抗IL-4和抗IL-13人源性单链抗体.     

Ovarian follicular concentration of IL-12, IL-15, IL-18 and p40 subunit of IL-12 and IL-23

BACKGROUND: The aim of the study was to determine the presence of interleukin (IL)-12, IL-15, IL-18 and p40 subunit of IL-12/IL-23 in follicular fluid from spontaneous cycles and the relation between the concentration of selected cytokines and IVF-embryo transfer outcome. METHODS: IVF-embryo transfer and enzyme immunoassay (EIA) (R&D Systems, Minneapolis, MN, USA and MBL, Nagoya, Japan) were used. RESULTS: Follicular fluid of women included in the IVF-embryo transfer procedure contained common p40 subunit of IL-12/IL-23 (median 70.1 pg/ml), IL-15 (median 1.3 pg/ml) and IL-18 (median 38.2 pg/ml). There was a significant negative correlation between follicular fluid concentrations of IL-15 and IL-18 (R=-0.392, P=0.003). Significantly higher concentrations of common p40 subunit of IL-12/IL-23 (median 79.8 pg/ml) were found in the follicular fluid taken from follicles containing oocytes, when compared with those without an oocyte (median 44.5 pg/ml, P=0.006). Patients who achieved clinical pregnancy had significantly decreased concentration of IL-15 (median 0.8 pg/ml) compared with patients without successful IVF-embryo transfer outcome (median 1.4 pg/ml, P=0.047). CONCLUSION: Follicular fluid collected from spontaneous cycles contains detectable levels of p40 subunit of IL-12/IL-23, IL-15 and IL-18. Increased concentrations of p40 subunit of IL-12/IL-23 in follicles containing oocytes suggest an important role of this cytokine in reproduction. Possible negative value of IL-15 as a predictor of IVF-embryo transfer success remains to be determined.     

Proinflammatory cytokines (IL-17, IL-6, IL-18 and IL-12) and Th cytokines (IFN-gamma, IL-4, IL-10 and IL-13) in patients with allergic asthma

Allergen-reactive T helper type-2 (Th2) cells and proinflammatory cytokines have been suggested to play an important role in the induction and maintenance of the inflammatory cascade in allergic asthma. We compared the plasma concentrations of novel proinflammatory cytokines IL-17 and IL-18, other proinflammatory cytokines IL-6 and IL-12, Th2 cytokines IL-10 and IL-13, and intracellular interferon-gamma (IFN-gamma) and IL-4 in Th cells of 41 allergic asthmatics and 30 sex- and age-matched health control subjects. Plasma cytokines were measured by enzyme-linked immunosorbent assay. Intracellular cytokines were quantified by flow cytometry. Plasma IL-18, IL-12, IL-10, IL-13 concentrations were significantly higher in allergic asthmatic patients than normal control subjects (IL-18: median 228.35 versus 138.72 pg/ml, P < 0.001; IL-12: 0.00 versus 0.00 pg/ml, P = 0.001; IL-10: 2.51 versus 0.05 pg/ml, P < 0.034; IL-13: 119.38 versus 17.89 pg/ml, P < 0.001). Allergic asthmatic patients showed higher plasma IL-17 and IL-6 concentrations than normal controls (22.40 versus 11.86 pg/ml and 3.42 versus 0.61 pg/ml, respectively), although the differences were not statistically significant (P = 0.077 and 0.053, respectively). The percentage of IFN-gamma-producing Th cells was significantly higher in normal control subjects than asthmatic patients (23.46 versus 5.72%, P < 0.001) but the percentage of IL-4 producing Th cells did not differ (0.72 versus 0.79%, P > 0.05). Consequently, the Th1/Th2 cell ratio was significantly higher in normal subjects than asthmatic patients (29.6 versus 8.38%, P < 0.001). We propose that allergic asthma is characterized by an elevation of both proinflammatory and Th2 cytokines. The significantly lower ratio of Th1/Th2 cells confirms a predominance of Th2 cells response in allergic asthma.     

IL-1, IL-18, and IL-33 families of cytokines

Summary: The interleukin-1 (IL-1), IL-18, and IL-33 families of cytokines are related by mechanism of origin, receptor structure, and signal transduction pathways utilized. All three cytokines are synthesized as precursor molecules and cleaved by the enzyme caspase-1 before or during release from the cell. The NALP-3 inflammasome is of crucial importance in generating active caspase-1. The IL-1 family contains two agonists, IL-1α and IL-1β, a specific inhibitor, IL-1 receptor antagonist (IL-1Ra), and two receptors, the biologically active type IL-1R and inactive type II IL-1R. Both IL-1RI and IL-33R utilize the same interacting accessory protein (IL-1RAcP). The balance between IL-1 and IL-1Ra is important in preventing disease in various organs, and excess production of IL-1 has been implicated in many human diseases. The IL-18 family also contains a specific inhibitor, the IL-18-binding protein (IL-18BP), which binds IL-18 in the fluid phase. The IL-18 receptor is similar to the IL-1 receptor complex, including a single ligand-binding chain and a different interacting accessory protein. IL-18 provides an important link between the innate and adaptive immune responses. Newly described IL-33 binds to the orphan IL-1 family receptor T1/ST2 and stimulates T-helper 2 responses as well as mast cells.     

IL-12 and IL-23 in health and disease

Food allergy affects up to 6% of children and 3–4% of adults in Westernized countries, and is the most common cause of outpatient anaphylaxis in most studies. The mainstay of treatment is strict avoidance of the offending allergens and education regarding the use of emergency medication in cases of accidental ingestions or exposures. While these approaches are generally effective, there are no definitive treatments that cure or provide long-term remission from food allergy. However, with recent advances in characterizing food allergens and understanding humoral and cellular immune responses in food allergy, several therapeutic strategies are being investigated. Potential treatments include allergen-specific immunotherapy as well as allergen-nonspecific approaches to downregulate the overall allergic response in food-allergic individuals.