de Carli M, D'Elios M M, Mariotti S, Marcocci C, Pinchera A, Ricci M, Romagnani S, del Prete G
Division of Clinical Immunology and Allergology, Institute of Clinica Medica 3, University of Florence, Italy.
J Clin Endocrinol Metab. 1993 Nov;77(5):1120-4. doi: 10.1210/jcem.77.5.8077301.
Lymphocytic infiltration of muscular and connective tissues of the retroorbital (RO) space is a histological hallmark of Graves' ophthalmopathy (GO). We have characterized some phenotypical and functional features of T cells derived from RO infiltrates of four GO patients who were submitted to orbital decompression. Fragments of RO tissue were cultured for 7 days in IL-2-conditioned medium in order to generate T cell lines of in vivo activated T cells. Phenotypical analysis of freshly isolated peripheral blood (PB) lymphocytes both from patients and four healthy controls showed a predominance of CD4+ T cells (CD4/CD8 ratios 1.9:2.5), whereas RO-derived T cell lines displayed almost equal proportions of CD4+ and CD8+ cells (CD4/CD8 ratios 0.9:1.2). RO T cell lines and PB T cells from patients and controls were then cloned using a high-efficiency cloning procedure. The phenotypical and functional features of 153 T cell clones (TCC) derived from RO infiltrates were examined and compared with those of 166 and 236 TCC derived from the PB of patients and controls, respectively. CD4/CD8 ratios ranged from 0.8-1.4 in the series of RO-derived TCC and from 1.9-2.2 in the corresponding series of PB-derived TCC. Assessment of lectin-dependent cytolytic activity showed similar proportions of cytotoxic clones in TCC derived from the PB of patients (37%) and controls (38%); most of the cytolytic TCC was CD8+. In contrast, the proportion of cytolytic RO TCC was markedly higher (106/153 = 69%), including 100% of CD8+ and the majority (59/79 = 75%) of CD4+ clones. When compared to TCC derived from the PB of both patients and controls, RO TCC showed remarkably high proportions of both CD8+ and CD4+ clones with a Th1-like cytokine profile, as documented by their ability to secrete IL-2, IFN-gamma, and tumor necrosis factor-alpha (TNF-alpha), but not IL-4 or IL-5. This study provides evidence that cytolytic T cells with Th1 profile of cytokine production predominate in RO infiltrates of GO, a pattern quite similar to those previously described in thyroid infiltrates of Hashimoto's thyroiditis or Graves' disease. The peculiar cytokine secretion profile of RO T cells may be of importance in the pathogenesis of both the tissue alterations and fibrogenic process observed in GO.
眼眶后（RO）间隙肌肉和结缔组织的淋巴细胞浸润是格雷夫斯眼病（GO）的组织学特征。我们已对4例接受眼眶减压的GO患者RO浸润部位来源的T细胞的一些表型和功能特征进行了鉴定。将RO组织碎片在白细胞介素-2条件培养基中培养7天，以生成体内活化T细胞的T细胞系。对患者和4名健康对照者新鲜分离的外周血（PB）淋巴细胞进行表型分析，结果显示CD4+ T细胞占优势（CD4/CD8比率为1.9:2.5），而RO来源的T细胞系中CD4+和CD8+细胞比例几乎相等（CD4/CD8比率为0.9:1.2）。然后，使用高效克隆程序对患者和对照者的RO T细胞系及PB T细胞进行克隆。检查并比较了153个RO浸润部位来源的T细胞克隆（TCC）与分别来自患者和对照者PB的166个及236个TCC的表型和功能特征。在RO来源的TCC系列中，CD4/CD8比率范围为0.8 - 1.4，在相应的PB来源的TCC系列中为1.9 - 2.2。凝集素依赖性细胞溶解活性评估显示，患者PB来源的TCC（37%）和对照者PB来源的TCC（38%）中细胞毒性克隆比例相似；大多数细胞溶解TCC为CD8+。相比之下，RO来源的细胞溶解TCC比例明显更高（106/153 = 69%），包括100%的CD8+克隆以及大多数（59/79 = 75%）的CD4+克隆。与患者和对照者PB来源的TCC相比，RO TCC显示出具有Th1样细胞因子谱的CD8+和CD4+克隆比例显著更高，这通过它们分泌白细胞介素-2、干扰素-γ和肿瘤坏死因子-α（TNF-α）的能力得到证明，但不分泌白细胞介素-4或白细胞介素-5。本研究提供了证据，表明具有Th1细胞因子产生谱的细胞溶解T细胞在GO的RO浸润中占主导，这种模式与先前在桥本甲状腺炎或格雷夫斯病的甲状腺浸润中描述的模式非常相似。RO T细胞独特的细胞因子分泌谱可能在GO中观察到的组织改变和纤维化过程的发病机制中具有重要意义。
