骨关节炎与免疫关联性初探

为了探讨免疫因素是否介导骨关节炎的发病 ,应用直接免疫荧光法检测了 6例晚期骨关节炎患者滑膜液中浸润淋巴细胞分化抗原的表达 ,同时用微量酶联免疫技术测定了这些患者滑膜液中IL 10、IL 12和sFasL的含量。结果显示关节滑膜液中的淋巴细胞以CD8+ 细胞为主 (6 7 0 %± 14 6 %) ,远远高于CD4+ 细胞 (15 0 %± 12 8%) ,两者相差显著 (P <0 0 1)。CD4+ /CD8+ 比值 <1。进一步分析表明其T细胞受体主要为αβ型TCR (6 4 0 %± 12 6 %)。滑膜液中IL 12含量 (2 0 3 84±49 2 )pg/ml高于IL 10 (37 8± 14 5 )pg/ml,有明显统计学差异 (P <0 0 1)。外周血中IL 10和IL 12的含量分别为 (4 3 37±14 2 )pg/ml和 (4 5 5 8± 10 6 )pg/ml。血清中sFasL含量较高 (15 3 90± 5 6 )pg/ml,而滑膜液中则很低 (<5pg/ml,P <0 0 1)。提示在骨关节炎的晚期CD8+ 淋巴细胞和IL 12增高可能参与了关节的免疫损伤。     

RA患者滑膜液及血清中IFN-γ、IL-10、IL-12与TRAIL的病理学意义

目的:研究IFN-γ、IL-10、IL-12及TRAIL在类风湿关节炎(RA)病理过程中的作用。方法:采用夹心ELISA法检测46名RA患者关节滑膜液和血清中IFN-γ、IL-10、IL-12及TRAIL的浓度。结果:RA患者关节滑膜液中IFN-γ、IL-12及TRAIL浓度(ng/L)分别为(106.2±7.8)、(57.7±3.5)和(166.5±12.3);RA患者血清中IFN-γ、IL-12及TRAIL浓度(ng/L)分别为(56.3±7.4)、(35.1±12.7)和(69.5±8.3);正常对照血清中IFN-γ、IL-12及TRAIL浓度(ng/L)分别为(31.1±4.4)、(25.2±2.6)和(41.2±4.8);RA患者关节滑膜液IFN-γ高于其血清含量(P<0.05)及正常对照组(P<0.01);RA患者关节滑膜液和血清中IL-10浓度和正常对照差异无统计学意义。结论:在RA疾病中IFN-γ和IL-12浓度明高于正常人,表明RA发病过程中细胞因子分泌格局以Th1型为主,并且滑膜液比外周血更为典型。RA患者关节滑膜液和血清中TRAIL浓度增高,说明RA疾病过程中伴随有凋亡分子的参与。     

类风关滑膜浸润T细胞特性与致病机制研究

为研究类风湿关节炎时关节滑膜浸润性T细胞生物学特性与致病机制 ,对 10例RA患者滑膜液中淋巴细胞的免疫表型、细胞因子分泌格局与趋化因子受体表达进行了分析。用双色荧光标记法分别测定滑膜液中和外周血淋巴细胞表型与趋化因子受体表达。用ELISA方法检测滑膜液与外周血中IFN γ、IL 10、IL 4与IL 12的含量。结果是滑膜液中的CD4 + T淋巴细胞为 4 0 0 %± 11% ,CD8+ T细胞为 34 0 %± 6 % ,CD4 + 与CD8+ T细胞的比值为 1 2 ,显著低于外周血中CD4 /CD8的比值。滑膜液中CD3和CD2 5双阳性的活化T细胞占 16 %± 6 0 %。趋化因子受体CCR5表达较低 ,与外周血无明显差异。但CX CR3表达水平较高 ,为 16 %± 4 0 % ,远远高于外周血 (仅为 0 5 %± 0 3% )。IFN γ在滑膜液中含量很高 ,达 (36 6 7± 4 3 2 )pg/ml,而外周血中含量仅为 (2 0 1± 3 2 )pg/ml。IL 4含量未能测得 (<15pg/ml ) ,与外周血相似。IL 12含量为 (4 19 9±89 2 )pg/ml,远高于外周血中的含量 (6 5 32± 34 2 )pg/ml。IL 10含量为 (187 7± 34 5 )pg/ml,高于外周血中的含量 (85±12 7)pg/ml。在所测细胞因子中 ,关节滑膜液中IFN γ和IL 12的含量与外周血相比具有显著的统计学差异。表明RA关节滑膜液中有相当数量的T细胞浸润。这些T细胞     

SLE患者IFN-γ、IL-4及IL-12水平变化及意义

从Th1/Th2细胞代表性细胞因子IFN γ/IL 4和IL 12水平的变化分析SLE患者Th1/Th2细胞的偏移现象。本实验通过半定量PCR法和ELISA酶联免疫检测法检测上述三种细胞因子含量。实验结果显示 ,SLE患者IL 4/IFN γ基因水平比值为0 32 8,高于对照组 (0 0 7)。ELISA检测显示IL 12在SLE患者血清中为 (38 39± 15 1)pg/ml,低于对照组 (84 97± 13 7)pg/ml(P <0 0 5 )。结论 :SLE患者Th1/Th2细胞因子平衡失调 ,IL 4细胞因子基因水平增高 ,同时血清中IL 12水平降低。     

类风湿性关节炎患者关节滑膜液浸润的T细胞表达特性

目的 :为研究类风湿性关节炎 (RA)患者关节滑膜液浸润的淋巴细胞介导自身免疫病的特性 ,分析了 2 2例RA患者滑膜液中淋巴细胞的免疫表型、对II型胶原的反应频率及IL 10、IL 12的分泌格局。方法 :用流式细胞术分别测定滑膜液和外周血淋巴细胞表型 ,并采用国际标准半有限稀释法分析了关节滑膜液中浸润的淋巴细胞对II型胶原 (CII)的反应频率 ,同时用ELISA方法检测了滑膜液与外周血中IL 10与IL 12含量。结果 :滑膜液中的T淋巴细胞的表型分别为CD4 (39 6 %±10 5 % )和CD8细胞 (36 4 %± 16 4 % ) ,CD4 CD8细胞比值显著低于外周血 ,且同时表达CD16和CD5 6的活化NK细胞占15 5 %± 11 1%。T细胞受体谱取用表明仍以αβTCR为主 (6 9 6 %± 15 7% )。有意义的是 :滑膜液中的T细胞对CII的反应频率为 15 2× 10 - 6 ,远远高于外周血 (4 0× 10 - 6 )。IL 12含量为 (4 19 9± 89 2 )pg ml,IL 10含量为 (187 7± 34 5 )pg ml,与外周血中这 2种细胞因子的含量〔分别为IL 12 :(6 5 32± 34 2 )pg ml和IL 10 :(85± 12 7)pg ml〕比较 ,具有显著的统计学差异。结论 :上述实验结果表明这种具有表达特性的浸润T细胞介导了RA患者关节滑膜组织的免疫损伤。     

IL—12以及IL—12P40对子宫内异症患者NK细胞功能的免疫调节

目的通过观察正常人及子宫内膜异位症(EM)患者外周血及腹腔液中IL-12、IL-12P40含量变化,分析其分子诱导NK细胞对子宫内膜细胞的细胞毒效应的影响,探讨IL-12、IL-12P40分子与EM患者NK细胞功能低下的相关性.方法ELISA酶联免疫检测IL-12、IL-12P40含量以及MTT释放法检测NK细胞毒活性.结果①EM患者腹腔上清液可使正常人外周血NK细胞杀伤活性(16.80%±3.6%)明显下降(8.37%±4.5% )(P＜0.05).② EM患者IL-12含量血清为66.38±12.6 pg/ml,低于对照组84.97±13.7 pg/ml(P＜0.05);腹腔液中为77.76±14.6 pg/ml,低于对照组106.92±10.7 pg/ml(P＜0.05).③EM患者IL-12P40含量血清为35.64±10.6 pg/ml, 与对照组无明显差异;腹腔液含量为79.76±12.6 pg/ml, 高于对照组40.54±10.0 pg/ml(P＜0.05), 并与疾病的程度呈负相关.④IL-12能增强NK细胞对子宫内膜细胞的杀伤,并呈剂量依赖.在10 ng/ml浓度诱导后杀伤率为31.90%,高于对照组的16.80%.⑤IL-12P40能拮抗IL-12对NK细胞活性的诱导,当加入IL-12P40后,使IL-12诱导的活性(29.3%)下降为19.36%.结论子宫内膜异位症患者腹腔液中IL-12P40含量的增高可能与NK细胞功能下降有关.     

子宫内膜异位症并不孕患者IL-10，IL13含量变化及其临床意义

目的检测IL-10和IL-13在子宫内膜异位症并不孕患者腹腔液及外周血中含量并探讨其意义。方法电视腹腔镜手术中采集EMS并不孕及对照组的腹腔液,术前空腹抽取外周血,采用ELISA法检测细胞因子水平。EMS并不孕组及对照组中,分别抽取38份腹腔液、外周血测IL-10及IL-13。结果EMS并不孕组和对照组腹腔液IL-10的含量分别为（15.8±12.9）pg/ml和（10.6±5.7）pg/ml,外周血中分别为（13.8±9.9）pg/ml和（8.6±5.7）pg/ml,两组比较均无统计学意义（P〉0.05）;EMS并不孕组和对照组腹腔液IL-13的含量分别为（93±9.6）pg/ml和（114±26）pg/ml,外周血中分别为（78±7.6）pg/ml和（102±29）pg/ml,两组比较均有统计学意义（P〈0.05）。结论异位子宫内膜细胞可抑制IL-13的产生,进而营造免疫抑制微环境,干扰免疫调节功能。     

较小剂量131Ⅰ联合中药治疗Graves病后血清sFas/sFasL变化及其与TRAb关系的探讨

目的探讨Graves病(GD)用较小剂量131Ⅰ联合中药治疗前后血清sFas/sFasL(可溶性细胞凋亡因子-1/sFas配体)变化的规律及其与TRAb的关系.方法31例GD患者,用较小剂量131Ⅰ(85.1～207.2MBq,平均140.6MBq)联合中药进行治疗,用酶联免疫吸附试验(ELISA)检测GD患者治疗前及治疗后不同时期血清sFas与sFasL含量,求出sFas/sFasL,并与正常对照组对比,分析GD患者血清sFas和sFasL含量,求出sFas/sFasL,并与正常对照组对比,分析GD患者血清sFas和sFasL与TRAb的相关关系.结果①GD患者治疗前sFas(179.8±64.2)pg/ml明显高于临床治愈期(104.2±23.5)pg/ml,P＜0.05和正常对照组(110.6±18.1)pg/ml,P＜0.05,甲低时的sFas(117.0±20.6)pg/ml与正常对照比无显著性差异.sFasL治疗前(353.8±92.8)pg/ml同样明显高于临床治愈期(261.1±85.3)pg/ml,P＜0.05和正常对照组(265.6±62.1)pg/ml,P＜0.05.甲低时的sFasL(257.1±55.3)pg/ml与正常对照比无显著性差异.②GD患者治疗前的sFas/sFasL(0.51±0.23)和甲低时(0.46±0.13)皆大于正常对照(0.41±0.16,P＜0.05).sFas和sFasL皆与TRAb呈正相关(r分别为0.52和0.49,P＜0.05).③较小剂量131Ⅰ联合中药治疗GD近期疗效良好,有效率100%(31/31),一次性治愈率74.2%(23/31),总治愈率83.9%(26/31).结论131Ⅰ联合中药治疗后sFas与sFasL呈规律性变化,治疗前和出现甲低时sFas呈优势表达,间接提示细胞凋亡与GD的治疗转归密切相关.TRAb可作为GD诊断和疗效观察的良好指标.     

白细胞介素-37在类风湿性关节炎患者血清表达水平的研究

目的 通过检测类风湿性关节炎(RA)患者血清中细胞因子白细胞介素(IL)-37、肿瘤坏死因子-α(TNF-α)、IL-18、炎性指标红细胞沉降率以及C-反应蛋白(C-reactionprotein,CRP)水平,观察RA患者临床数据包括压痛关节数,肿胀关节数以及DAS28评分等,探讨RA患者血清IL-37水平升高的意义以其在RA发病机制中可能的作用.方法 80例RA患者、80例健康对照患者,通过酶联免疫吸附试验(ELISA)法检测其血清中细胞因子水平.结果 RA患者血清中IL-37[(40.33±11.25)pg/mL]、TNF-α[(110.41 ±35.37) pg/mL]、IL-18[(121.73±29.22) pg/mL]水平以及红细胞沉降率(ESR)[(42.31±15.02) mm./h]、CRP[(38.31±17.22) mg/L]水平明显高于对照组IL-37[(18.21±5.72) pg/mL]、TNF-α[(30.19±6.82) pg/mL]、IL-18[(55.47±7.29) pg/mL]水平,组间差异有统计学意义(P＜0.05).IL-37的表达与TNF-α、IL-18水平呈正相关(相关系数r=0.981,P=0.001).结论 IL-37在RA患者体内高表达并与其它几种炎性因子的表达具有相关性,IL-37可能作为炎性抑制因子参与了RA的发生、发展.     

特异性免疫治疗对哮喘和鼻炎患者血清炎性细胞因子的影响

目的通过检测对屋尘螨过敏的的哮喘和/或鼻炎患者在特异性免疫治疗或常规药物治疗过程中血清白介素(IL)-5、IL-10、IL-17、INF-γ及TNF-α等5种细胞因子的水平,探讨特异性免疫治疗对过敏性哮喘和鼻炎患者血清炎性细胞因子的影响。方法选取44例在广州呼吸疾病研究所接受标准化屋尘螨特异性免疫治疗(SIT)满1年的哮喘和/或鼻炎患者作为SIT组,50例对屋尘螨过敏并未接受SIT治疗的哮喘和/或鼻炎患者作为病例组,另外选取23例无过敏、无哮喘与鼻炎的健康自愿者作为对照组,用悬浮芯片方法检测三组人群血清中IL-5、IL-10、IL-17、INF-γ及TNF-α等5种细胞因子的水平。结果服从正态分布的数据以均数±标准差表示,偏态分布的数据以中位数(四分位间距数)表示。FEV1%(%)在SIT组、病例组及对照组中分别为93.05±15.62、87.0±13.51、95.51±7.84,三组比较显示无显著差异(P>0.05);AHR在这三组中分别为2(0,3)、1(0,2)、0(0,0),三组比较显示SIT组及病例组均显著高于对照组(P<0.05),SIT组与病例组比较无显著差异(P>0.05);血清IL-5(pg/mL)在SIT组、病例组及对照组中的水平分别为1.46(0.62,2.71)、1.95(1.18,2.46)、0.34(0.01,0.83),其中SIT组显著低于病例组(P=0.001),病例组显著低于对照组(P<0.001);IL-10(pg/ml)在SIT组、病例组及对照组中的水平分别为1.6(0.95,2.69)、1.25(0.93,1.8)、0.43(0.21,1.07),SIT组与病例组显著高于对照组(P<0.001);TNF-α(pg/mL)在SIT组、病例组及对照组中的水平分别为10.81(2.18,31.3)、3.39(1.78,6.83)、0.09(0.09,1.18),其中SIT组显著高于病例组与对照组(P=0.004),病例组又显著高于对照组(P<0.001)。IL-17与INF-γ在三组受试者的血清中均不可测出。结论哮喘和鼻炎患者血清中IL-5、TNF-α水平显著高于正常人,说明IL-5和TNF-α在哮喘和/或鼻炎的致病中发挥重要作用;关于SIT对炎性细胞因子的影响还有待我们进一步的研究。     

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.     

Selective activation of mast cells in rheumatoid synovial tissue results in production of TNF-α, IL-1β and IL-1Ra

Objectives and Design: To study the consequences of mast cell activation in human synovial tissue. Methods: Synovial tissue was obtained from 18 RA patients and mast cells was selectively activated in synovial tissue explant cultures. Expression of TNF-α, IL-1β and IL-1Ra were determined and tissue distribution of IL-1β was studied. Results: Compared to untreated synovia, selective activation of synovial mast cells increased significantly the production of TNF-α (0.49 ± 0.88 vs. 4.56 ± 3.18 pg/mg wet tissue, p < 0.001) and IL-1β (0.058 ± 0.032 vs. 2.55 ± 1.98 pg/mg wet tissue, p = 0.013). The expression of TNF-α and IL-1β mRNA increased significantly (19-fold (p = 0.009) and 13-fold (p = 0.031), respectively). Mast cell activation induced IL-1β expression in particular in nearby CD68 positive synovial macrophages. Secretion of IL-1Ra was also increased but to a lesser degree than that of IL-1β. Conclusions: Synovial mast cells produce proinflammmatory cytokines and may thus contribute to the inflammation in RA. Received 21 August 2006; returned for revison 13 October 2006; accepted by J. Di Battista 8 November 2006     

Increased IL-2, IL-4 and interferon-gamma (IFN-gamma) in steroid-sensitive nephrotic syndrome.

We investigated the production of cytokines by peripheral blood mononuclear cells (PBMC) and serum cytokine concentrations in children with steroid-sensitive idiopathic nephrotic syndrome (SSNS). PBMC from patients off treatment were collected during remission and relapse and cultured in medium alone or stimulated with calcium ionophore plus phorbol myristate acetate. Control PBMC were taken from healthy age-matched children. IL-2 was measured by bioassay, IL-4 by immunoradiometric assay, and IL-8 and IFN-gamma by ELISA. After 24 h culture without stimulation, IL-2, IL-4 and IFN-gamma were not detectable in the supernatant in any of the children. After stimulation, the supernatant concentrations of IL-2 (median 172 U/ml at 24 h) and IL-4 (160 pg/ml at 24 h; 210 pg/ml at 72 h) were significantly increased in relapse compared with remission (IL-2 37 U/ml; IL-4 65 pg/ml and 60 pg/ml) and controls (IL-2 69 U/ml; IL-4 40 pg/ml and 40 pg/ml) (P < 0.05). The concentration of IFN-gamma was not significantly increased in relapse compared with remission and controls (600, 325, and 145 U/ml, respectively, at 72 h). IL-8 concentrations were similar in relapse, remission and controls with stimulation (median 32, 40 and 40 ng/ml, respectively) and without (30, 17 and 10 ng/ml). IL-2 was not detectable in serum, but IL-4, IL-8 and IFN-gamma were measurable in about half the patients, both in relapse and remission, though were virtually undetectable in controls. We conclude that relapse of SSNS in children is associated with T lymphocyte activation with release of IL-2, IL-4 and IFN-gamma.     

Profile of Cytokines in Synovial Fluid Specimens from Patients with Arthritis. Interleukin 8 (IL-8) and IL-6 Correlate with Inflammatory Arthritides

The synovial fluid aspirated from patients with symptomatic arthritis was analyzed for the presence of tumor necrosis factor (TNF), interleukin 6 (IL-6) and interleukin 8 (IL-8). All three cytokines were found in both inflammatory and non-inflammatory arthritides: IL-8 levels ranged from less than 20 to 38,990 pg/ml, IL-6 from less than 10 to 72,300 pg/ml and TNF from less than 4 to 61 pg/ml. No inhibitors of cytokine activity were found. IL-8 and IL-6 were present in significantly higher levels in patients with inflammatory arthritis compared to patients with osteoarthritis, and there was significant correlation between the IL-6 and IL-8 levels. These findings document the presence of multiple cytokines in the synovial fluid specimens of patients with arthritis, and demonstrate that higher cytokine levels accompany inflammatory arthritis.     

Interrelationships between interleukin (IL)-1, IL-6 and IL-8 in synovial fluid of various arthropathies

High levels of many cytokines, including interleukin (IL)-1, IL-6 and IL-8, were found in various arthropathies suggesting that they play a role in the pathogenesis of disease, although their relationship with the type and activity of disease is still not clear. The synovial fluid (SF) of 24 patients with rheumatoid arthritis (RA), 19 with psoriatic arthritis (PA) and 33 with osteoarthritis (OA) was analyzed for IL-1, IL-6 and IL-8. The highest concentration of the three cytokines was found in the SF of RA. IL- detectable levels (>-20 pg/ml) were observed in 8/24 (33.3%) patients with RA, in one patient with PA but in no patient with OA.IL-6 (mean±SD) (1610.37±1781.65 pg/ml) was higher in RA than in PA (672.47±867.40 pg/ml,p=0.043) and OA (89.45±120.52 pg/ml,p=0.0001). IL-8 (1042.72±698.64 pg/ml) was higher in RA than in PA (660.36±625.11 pg/ml,p=0.03) and OA (89.9±45.88 pg/ml,p=0.0001). A correlation between IL-1, IL-6 and IL-8 was found in RA. In all patients a correlation between IL-6 and IL-8 levels was found; moreover, these two cytokines were associated with SF indices of inflammation, such as white blood cells (WBC) count and total protein (TP) concentration.Out findings suggest that these interrelationships play a role in the evolution of more severe erosive arthropathy such as RA.