丝氨酸蛋白酶3及蛋白酶活化受体-2激动剂对韦格纳肉芽肿病患者树突状细胞样单核细胞的影响

目的 探索丝氨酸蛋白酶3(PR3)对人外周血树突状细胞(DC)样单核细胞-CD14+CD16high单核细胞的蛋白酶活化受体(PAR)-2表达、成熟及细胞因子产生的作用.方法 密度梯度离心法分离健康人和韦格纳肉芽肿病(WG)患者外周血单个核细胞(PBMC);分别以脂多糖、PR3、胰蛋白酶、PAR-2激动肽(PAR-2-AP)、脂多糖+PR3、脂多糖+胰蛋白酶在体外刺激PBMC 24h;流式细胞仪检测刺激后的CD14+CD16high单核细胞表面PAR-2及CD80、CD83、人类白细胞抗原(HLA)-DR的表达;酶联免疫吸附试验(ELISA)检测培养上清中细胞因子白细胞介素(IL)-6的分泌.统计学分析采用Mann Whitney 非参数检测方法.结果 PR3、胰蛋白酶和PAR-2-AP对外周血DC样单核细胞表面PAR-2和CD80、CD83、HLA-DR表达均无影响;脂多糖能够显著升高健康人细胞表面PAR-2[(5.8±1.5)%升至(24.5±4.5)%,P=0.002]及WG患者和健康人CD80、CD83、HLA-DR的表达;PR3、胰蛋白酶、PAR-2-AP及脂多糖均能诱导IL-6的分泌.结论 PR3和PAR-2通路激动剂不能诱导DC样单核细胞的PAR-2的表达及成熟,但能诱导IL-6分泌.

Abstract：

Objective To assess the effects of protease 3(PR3)and protease-activated receptor (PAR)-2-activator on the maturation and functions of peripheral blood dendritic cell(DC)-like monocytes.Methods Density gradient centrifugation was used to isolate peripheral blood mononuclear cells(PBMC)from Wegener's granulomatosis(WG)patients and healthy controls(HC).PBMC were stimulated by LPS,human PR3,trypsin,PAR-2-agonist peptide (PAR-2-AP),LPS+PR3 or LPS+trypsin for 24 h.Flow cytometry was used to analyze the expression of PAR-2,CD80,CD83,HLA-DR on stimulated DC-like monocytes-CD14+CD16high monocytes.ELISA kit was used to test the concentration of IL-6 in the culture supernants.Mann-Whitney non-parameteric test was used fur statistical analysis.Resuits No effect of PR3,trypsin and PAR-2-AP on the expression of PAR-2,CD80,CD83,HLA-DR of DC-like monoeytes was found.LPS could significantly induce PAR-2 expression in HC[from(5.8±1.5)%to(24.5±4.5)%,P=0.002]and the expression of CD80,CD83,HLA-DR in HC and WG;PR3,trypsin,PAR-2-AP and LPS could all stimulate the secretion of IL-6.Conclusion PR3 and PAR-2 pathway-activators can not promote PAR-2expression and maturation of DC-tike monocytes,but they can induce the secretion of IL-6.     

韦格纳肉芽肿

韦格纳肉芽肿又称韦格纳肉芽肿病 ( WG)。 193 6年首先由 Wegener描述。主要表现为呼吸道肉芽肿性坏死性血管炎及弥漫性坏死性血管炎 ,肾小球肾炎常见 ,为一种少见严重的多系统受累疾病。WG原因不明 ,早期病变内常有急性白细胞破碎性血管炎的改变 ,以后发现 Ig G、Ig A及 CIC异常。近年发现抗中性粒细胞浆抗体与 WG相关 ,这些都支持本病为一种自身免疫性疾病。WG在白种人多见 ,在美国估计其发病率为百万分之三。多见于男性 ,常起病于 3 0～ 4 0岁。少数患者起病急骤 ,多数呈现慢性病程。本病无特异的前驱症状 ,病情呈进行性发展。常…     

韦格纳肉芽肿一例

患者,男性,58岁,因咳嗽、咳痰2个月,右胸痛,关节痛1个月入院.2个月前受凉后出现咳嗽、咳少量黄痰,自服感冒药等疗效欠佳.1个月前无明显诱因出现右侧胸背部疼痛,伴肩、肘、腕、踝等关节疼痛,到外院诊治,行肺CT检查发现两肺多发性团块、结节,考虑炎性假瘤可能,予抗感染及对症支持治疗半个月,病情无明显好转,为求进一步诊治而到我院呼吸科就诊.     

误诊为韦格纳肉芽肿病的T细胞恶性淋巴瘤一例

韦格纳肉芽肿病 (Wegener′sgranulomatosus,WG)是肉芽肿性坏死性血管炎 ,以主要累及上、下呼吸道和肾脏为其特点。现将 1例误诊为WG的累及上呼吸道和肾脏的T细胞恶性淋巴瘤报道如下。患者 :男性 ,2 2岁。因反复鼻塞、脓涕 8年 ,加重伴发热1个月于 2 0 0 3年 6月 17日入院。 1995年开始出现反复鼻塞、流脓涕 ,偶有涕中带血。 1998年于当地医院查为“双侧鼻息肉” ,行手术摘除 (无病理资料 ) ,术后出现高热 ,体温最高达 4 1℃ ,经病原学检查为“阴沟杆菌、霉菌”感染 ,用抗感染治疗后缓解。此后鼻塞、脓涕仍时有发作。 5月 10日无明显诱因…     

韦格纳肉芽肿病并发肾动脉瘤2例及文献复习

目的总结分析韦格纳肉芽肿病(Wegener's granulomatosis,WG)合并肾动脉瘤患者的临床特征和治疗预后。方法报道本院2例WG合并肾动脉瘤,并复习文献总结其临床特征、治疗及预后。结果综合文献报道WG合并动脉瘤共计15例(包括本文2例),其中男性14例,所有患者均有上呼吸道、肺和肾脏受累。13例动脉瘤出现在发病初期,并发动脉瘤最具提示意义的症状为腹痛;15例患者中4例累及大动脉(主动脉),11例累及中等动脉,其中肾动脉分支受累7例;9例发生动脉瘤破裂,其中3例死于腹腔大出血。肾动脉瘤破裂临床表现为腹痛和出血性贫血,腹部B超和CT提示肾周血肿,肾动脉造影可见多发动脉瘤;在联合应用大剂量肾上腺糖皮质激素和环磷酰胺的基础上,弹簧栓塞破裂的肾动脉以及输血支持治疗可挽救患者生命。结论韦格纳肉芽肿病合并肾动脉瘤为罕见但危及生命的并发症。WG患者出现腹痛时要警惕肾动脉瘤形成或破裂的可能,及时行肾动脉造影有利于明确诊断;破裂动脉瘤可予弹簧栓塞等处理,同时联合大剂量糖皮质激素和环磷酰胺治疗,以改善预后。     

韦格纳肉芽肿病T细胞表达及分泌细胞因子的研究—Th1型免疫反应

目的：通过分析Ｔ细胞细胞因子的表达和分泌，探讨韦格纳肉芽肿病（ＷＧ）的免疫反应类型。方法：用有限稀释法从ＷＧ患者鼻粘膜活检标本（ＮＢＳ）、支气管肺泡灌洗液（ＢＡＬ）及外周血中克隆出Ｔ细胞。用ＲＴＰＣＲ及ＥＬＩＳＡ检测其细胞因子的表达和分泌。结果：所有７株从ＮＢＳ克隆的Ｔ细胞只表达和分泌Ｔｈ１型细胞因子（ＩＦＮγ）。从ＢＡＬ克隆的３株Ｔ细胞，２株为Ｔｈ１型，另１株为Ｔｈ０型（表达和分泌ＩＦＮγ及ＩＬ４）。而从外周血中克隆的２７株Ｔ细胞。Ｔｈ１型（９株）多于Ｔｈ２型（２株），但多数（１６株）为Ｔｈ０型。另外，ＷＧ患者外周血中未分类的多克隆ＣＤ＋４和ＣＤ＋８Ｔ细胞产生ＩＦＮγ的量明显高于正常对照者，而ＩＬ４表达和分泌则与正常无差别。结论：ＷＧ的免疫病理过程主要表现为Ｔｈ１型免疫反应     

“韦格纳肉芽肿”重新命名

2011年,美国风湿病学会、美国肾脏病学会及欧洲风湿病学会联合提出将"韦格纳肉芽肿"更名为"肉芽肿性多血管炎",本文对于该疾病命名的历史沿革,病理学特征及与其他血管炎的关系进行论述,新命名更能反映出本病的临床和病理生理学特征,指引研究方向。     

重视韦格纳肉芽肿病的诊断与治疗

血管炎病作为一大类风湿性疾病 ,在我国很长一段时间内未被大多数临床医师所认识 ,其临床的复杂性和诊治的困难性也一直是困扰我们风湿病专科医师的一个问题。近几年由于风湿病学在我国的普及发展 ,血管炎病已逐渐引起人们的重视。张乃峥教授于本刊 1998年第 3期发表了“由三个个案报道引起的思考———论血管炎病” ,提出要重视血管炎病的诊断与治疗。去年中华医学会风湿病学分会召开了全国血管炎的专题研讨会 ,这些促进了人们对血管炎病的认识。韦格纳肉芽肿病 (Wegener′sgranulomatosis ,WG)是一种坏死性肉芽肿性血管炎 ,近几年在我国…     

Endothelial protease-activated receptor-2 induces tissue factor expression and von Willebrand factor release

A second protease-activated receptor (PAR-2) that could be activated by trypsin or more physiologically by mast cell tryptase has been recently cloned. Both the structure and activation mechanism of PAR-2 was similar to the functional thrombin receptor (PAR-1). Although many effects of the coagulation protease thrombin on the vascular endothelium could be attributed to PAR-1 activation, very little is known about the physiological and pathophysiological role of PAR-2. We investigated whether stimulation of PAR-2 on endothelial cells induced two cellular responses that play a central role in primary and secondary haemostasis: the release of high molecular weight von Willebrand factor (hmw-VWF) from Weibel-Palade bodies and the de novo synthesis of tissue factor (TF) mRNA and protein. Human umbilical vein endothelial cells (HUVEC) were incubated with agonists for PAR-2 at 37 degrees C. Both trypsin and SLIGKV increased TF mRNA and activity and induced the release of hmw-VWF due to elevated levels of cytosolic Ca2+. Trypsin (10 nm) induced a 6-fold increase of TF mRNA and reduced time until fibrin clot formation to 37%, indicating trebling of the cell surface located TF activity. Stimulation of HUVEC with the PAR-2 agonist peptide SLIGKV induced a dose-dependent increase of TF mRNA up to 6 times and TF activity up to 3 times. Release of hmw-VWF was achieved both after incubation of HUVEC with trypsin and SLIGKV and was directly depending on intracellular Ca2+ mobilization. To make results comparable to the functional thrombin receptor, homologous experiments were carried out using the PAR-1 agonists thrombin and SFLLRN.     

Protease-activated receptor-2 antagonists and agonists

Interest in the development of specific antagonists of the protease-activated receptors are significant, however, achieving such goals remain extremely challenging. Considerable efforts have been directed at developing specific antagonists of the first elucidated member of this receptor family, namely the thrombin receptor, PAR-1. However, significantly less effort has been directed at the second member of the family, PAR-2 due in part to lack of clarity concerning its activating protease(s), and uncertainty concerning its physiological and pathophysiological roles in disease pathways. This review will briefly summarize what is known about the activating protease(s), the potential (patho)physiological roles for PAR-2 and structure-activity relationships that have been developed for PAR-2 agonists and antagonists in relationship to agonists and antagonists developed for the other protease-activated receptors.     

Limited versus severe Wegener's granulomatosis: baseline data on patients in the Wegener's granulomatosis etanercept trial

OBJECTIVE: To report baseline data on 180 patients with Wegener's granulomatosis (WG) enrolled in the WG Etanercept Trial (WGET), and to examine demographic and clinical differences between patients with limited disease and those with severe disease. METHODS: Definitions of limited and severe disease were agreed upon by consensus of the investigators at a pretrial meeting and were incorporated into the protocol as a stratification criterion. These data were applied prospectively to the WGET patient cohort, based on clinical features and the intention to treat patients according to disease activity. Data related to disease onset, date of diagnosis, clinical features, antineutrophil cytoplasmic antibody assays, tissue biopsy findings, and other medical history were collected on a baseline medical history form. Physician-investigators from each center participated in the development of this form, and all were certified in its use prior to the start of the trial. Selected data on patients who were screened for the trial but were not enrolled were also collected. RESULTS: Several significant differences between the limited and severe disease subsets were observed. Patients with limited disease were nearly a decade younger at disease onset compared with patients with severe disease. Thirty-three percent of patients with severe disease were women, compared with 58% of those with limited disease. Despite their younger age at symptom onset, patients with limited disease tended to have longer disease duration, a greater likelihood of experiencing exacerbation of previous disease following a period of remission, and a higher prevalence of destructive upper respiratory tract disorders at the time of enrollment (e.g., saddle-nose deformity). Patients with limited WG were less likely than those with severe disease to have antibodies to either proteinase 3 or myeloperoxidase. Patients with severe disease had a higher likelihood of previous thyroid disease, particularly either Graves' disease or Hashimoto thyroiditis, suggesting the possibility of different pathogenetic factors within these disease subsets. Other observed differences between these subsets, such as the greater frequency of alveolar hemorrhage in the severe disease group, were related to the a priori definitions of limited and severe disease. CONCLUSION: There are significant differences between patients with limited WG and those with severe WG with regard to sex, age, the likelihood of recurrent disease, the risk of damage in certain organ systems, and, possibly, etiologic factors. These differences (and perhaps other differences that are currently unrecognized) in patient subsets may have implications for mechanisms of pathogenesis, prognosis, response to treatment, and the design of future clinical investigations.     

Cardiac involvement in patients with Wegener's granulomatosis

Wegener's granulomatosis (WG) is a systemic necrotizing vasculitis, which could potentially affect any organ system. However, there have only been a few reports on cardiac involvement. We described the echocardiographic findings in nine patients affected by WG. A complete M-mode, two-dimensional, Doppler and color-Doppler transthoracic echocardiogram was performed in nine patients (seven females and two males) affected by WG. In each patient, cardiac abnormality, for example, valvular damage, left ventricular global systolic dysfunction, or pericardial effusion, was detected. In particular, heart valve disease was found in eight patients, and in three cases, aortic valve insufficiency, which was severe enough to require surgical valve replacement, was observed. Cardiac involvement in patients with WG is common. In particular, there is a high frequency of aortic valve abnormalities. Thus, an echocardiographic study should be routinely performed. Received: 21 October 1999 / Accepted: 28 April 2000     

Accelerated atherosclerosis in patients with Wegener's granulomatosis

BACKGROUND: Several autoimmune disorders are complicated by excess cardiovascular disease. In addition to traditional risk factors, non-traditional risk factors such as endothelial activation and excessive vascular remodelling might be determinants of the progression of atherosclerosis in patients with an autoimmune disease. OBJECTIVE: To evaluate whether patients with Wegener's granulomatosis (WG) have an increased prevalence of atherosclerosis and to determine predisposing factors. METHODS: 29 WG patients (19 men; mean (SD) age, 53 (14) years) with inactive disease and 26 controls (16 men; age 53 (15) years) were studied. Common carotid intima-media thickness (IMT) was measured by ultrasound. In all individuals traditional risk factors for cardiovascular disease were determined. High sensitivity C reactive protein (hsCRP) was measured. Endothelial activation was assessed by measuring thrombomodulin, vascular cell adhesion molecule-1, and von Willebrand factor. As a marker of vascular remodelling matrix metalloproteinases (MMP-3 and MMP-9) and TIMP-1 were measured. RESULTS: IMT was increased in WG patients compared with controls (p<0.05). No differences in traditional risk factors and endothelial activation markers between patients and controls were found. Levels of hsCRP, MMPs, and TIMP-1 were increased in WG patients (p<0.05). CONCLUSIONS: Increased IMT found in WG patients cannot be explained by an increased prevalence of traditional risk factors. Although endothelial activation markers in WG patients with inactive disease were not increased, the raised levels of hsCRP, MMPs, and TIMP-1 suggest that enhanced inflammation and excessive vascular remodelling are contributing factors in the development of accelerated atherosclerosis in WG.     

Wegener's granulomatosis in patients with rheumatoid arthritis

OBJECTIVE: To describe 2 cases of coexisting rheumatoid arthritis (RA) and Wegener's granulomatosis (WG), and to summarize the clinical and serological data for all 6 patients reported in the English literature since 1966. METHODS: Medline review over a 35-year period (1966-2002) revealed 4 reported cases of RA associated with WG. Patients were diagnosed based on symptoms, radiographic changes, bronchoalveolar lavage fluid analysis, hematuria, serology, and biopsy. We describe 2 additional cases of WG developing in Caucasian women with RA. These are the first reported patients to possess positive antineutrophil cytoplasmic antibodies (ANCA) and autoantibodies to proteinase 3 (PR3). RESULTS: All 6 cases of coexisting RA and WG were female. The diagnosis of RA preceded WG diagnosis in all cases; mean age at RA onset was 43.7 +/- 15.0 years, duration of RA prior to WG diagnosis 7.9 +/- 9.1 years. Clinical findings included erosive articular disease on radiographs (n = 4; 67%), positive rheumatoid factor (n = 6; 100%), upper respiratory involvement (n = 5; 83%), lower respiratory signs (n = 4; 67%), renal involvement (n = 2; 33%), and positive ANCA (n = 2/3; 67%). Five patients were treated with corticosteroids and cyclophosphamide, with clinical improvement. CONCLUSION: Although rare, WG may develop in patients with preexisting RA and may present with end-organ involvement.     

Monocyte activation in patients with Wegener's granulomatosis

OBJECTIVE: Wegener's granulomatosis (WG) is an inflammatory disorder characterised by granulomatous inflammation, vasculitis, and necrotising vasculitis and is strongly associated with anti-neutrophil cytoplasmic antibodies (ANCA). Activated monocytes/macrophages are present in renal biopsy specimens and participate in granuloma formation by synthesising and secreting a variety of chemoattractants, growth factors, and cytokines. In view of these findings, in vivo monocyte activation was evaluated in patients with WG and the findings related to parameters of clinical disease activity. METHODS: Monocyte activation was analysed by measuring plasma concentrations of soluble products of monocyte activation, that is neopterin and interleukin 6 (IL6), by ELISA, and by quantitating the surface expression of activation markers on circulating monocytes by flow cytometry. RESULTS: Twenty-four patients with active WG were included in this study. Ten of these patients were also analysed at the time of remission. Twelve patients with sepsis served as positive controls, and 10 healthy volunteers as negative controls for monocyte activation. Patients with active disease had increased monocyte activation compared with healthy controls as shown by increased concentrations of neopterin (p < 0.0001) and increased surface expression of CD11b (p < 0.05) and CD64 (p < 0.05). In those patients with increased concentrations of IL6 during active disease plasma concentrations of IL6 decreased during follow up when patients went into remission (p < 0.0001). In addition, neopterin (r = 0.37, r = 0.44), IL6 (r = 0.37, r = 0.60) and CD63 expression (r = 0.39, r = 0.45) correlated significantly with disease activity as measured by the Birmingham Vasculitis Activity Score and C reactive protein values, respectively. Compared with patients with sepsis, all markers of monocyte activation in patients with vasculitis were lower. CONCLUSION: It is concluded that disease activity in WG correlates with the extent of activation of monocytes, compatible with their role in the pathophysiology of this disease.