Noronha I L, Krüger C, Andrassy K, Ritz E, Waldherr R
Department of Pathology, Ruperto-Carola University, Heidelberg, Germany.
Kidney Int. 1993 Mar;43(3):682-92. doi: 10.1038/ki.1993.98.
Humoral and cellular immune mechanisms are thought to be involved in various forms of vasculitis and glomerulonephritis. Recent clinical and experimental results point to a role of cytokines in ANCA-positive vasculitides. We analyzed tumor necrosis factor-alpha (TNF-alpha), interleukin-1 beta (IL-1 beta) and interleukin-2 receptors (IL-2R) in renal biopsies and in plasma from 22 patients with Wegener's granulomatosis and microscopic polyangiitis. Kidney biopsies were examined by immunocytochemistry, polymerase chain reaction and in situ hybridization. Immunoreactive TNF-alpha, IL-1 beta and/or IL-2R positive infiltrating cells were observed in 21 of 22 biopsies. TNF-alpha, IL-1 beta and IL-2R staining was evident in the interstitium and at periglomerular and perivascular sites. The number of positive cells was markedly increased in biopsies with active lesions. Positive cells were also present in cellular and fibrocellular crescents, surrounding tuft necrosis and in the walls of arteries and arterioles with acute vasculitic lesion. Some tubular epithelial cells stained for TNF-alpha and IL-1 beta. TNF-alpha, IL-1 beta and IL-2R positive infiltrating cells correlated with the presence of histologically active renal lesions. The evaluation of TNF-alpha and IL-1 beta expression at the mRNA level assessed by the polymerase chain reaction demonstrated specific transcripts for TNF-alpha and IL-1 beta in all six cases analyzed. In situ hybridization studies showed an increased expression of mRNA for TNF-alpha and IL-1 beta in infiltrating mononuclear cells, in epithelial cells of Bowman's capsule and in some tubules, predominantly of patients with active renal lesions. The results at the mRNA level correlated with the immunocytochemical findings. Compared to healthy individuals higher TNF-alpha plasma levels were observed in patients with vasculitis (34.4 +/- 16.6 pg/ml (SEM) vs. 1.9 +/- 0.7 pg/ml in controls; P < 0.01). All patients presented a marked increase in sIL-2R plasma levels (3512 +/- 485 U/ml vs. 397 +/- 21 U/ml in healthy controls; P < 0.001). IL-1 beta was not detected in most plasma samples. Elevated TNF-alpha and sIL-2R plasma levels were related to active renal lesions. Our study clearly demonstrates that in ANCA-positive vasculitis TNF-alpha and IL-1 beta are produced in situ by activated infiltrating mononuclear cells and resident renal cells. Intrarenal localization of cytokine producing cells and the correlation between cytokine production and histological signs of activity suggest that TNF-alpha and IL-1 beta are important locally acting mediators in the vasculitic/glomerulonephritic process.
体液免疫和细胞免疫机制被认为参与了各种形式的血管炎和肾小球肾炎。最近的临床和实验结果表明细胞因子在抗中性粒细胞胞浆抗体（ANCA）阳性血管炎中发挥作用。我们分析了22例韦格纳肉芽肿病和显微镜下多血管炎患者肾活检组织及血浆中的肿瘤坏死因子-α（TNF-α）、白细胞介素-1β（IL-1β）和白细胞介素-2受体（IL-2R）。通过免疫细胞化学、聚合酶链反应和原位杂交对肾活检组织进行检查。在22例活检组织中的21例观察到免疫反应性TNF-α、IL-1β和/或IL-2R阳性浸润细胞。TNF-α、IL-1β和IL-2R染色在间质、肾小球周围和血管周围部位明显。在有活动性病变的活检组织中，阳性细胞数量明显增加。在细胞性和纤维细胞性新月体、肾小球节段坏死周围以及有急性血管炎病变的动脉和小动脉壁中也存在阳性细胞。一些肾小管上皮细胞TNF-α和IL-1β染色阳性。TNF-α、IL-1β和IL-2R阳性浸润细胞与组织学上有活性的肾脏病变的存在相关。通过聚合酶链反应评估的mRNA水平上TNF-α和IL-1β表达的研究表明，在所分析的所有6例病例中均有TNF-α和IL-1β的特异性转录本。原位杂交研究显示，在浸润的单核细胞、鲍曼囊上皮细胞和一些肾小管中，TNF-α和IL-1β的mRNA表达增加，主要见于有活动性肾脏病变的患者。mRNA水平的结果与免疫细胞化学结果相关。与健康个体相比，血管炎患者血浆中TNF-α水平更高（34.4±16.6 pg/ml（标准误），对照组为1.9±0.7 pg/ml；P<0.01）。所有患者血浆中可溶性IL-2R（sIL-2R）水平均显著升高（3512±485 U/ml，健康对照组为397±21 U/ml；P<0.001）。大多数血浆样本中未检测到IL-1β。血浆中TNF-α和sIL-2R水平升高与活动性肾脏病变相关。我们的研究清楚地表明，在ANCA阳性血管炎中，TNF-α和IL-1β由活化的浸润单核细胞和肾脏固有细胞原位产生。细胞因子产生细胞的肾内定位以及细胞因子产生与活动组织学征象之间的相关性表明，TNF-α和IL-1β是血管炎/肾小球肾炎过程中重要的局部作用介质。
