muramidase and Graves-Ophthalmopathy

Graves' orbitopathy (GO) is an autoimmune inflammatory ocular complication and one of the most frequent manifestations of Graves' disease (GD). Clinical judgment of GO is subjective sometimes leading to clinical and therapeutic challenges. Better tools to diagnose this severe complication are warranted.. The aim of the present study was to evaluate tear levels of LYZ, LACRT and AZGP1 in GD patients with or without GO, as possible biomarkers for GO. Tear samples were collected from GD patients with moderate-to-severe GO (n = 21) and no clinical signs of GO (n = 21). Additionally, 18 GD patients with mild GO and 9 patients without GO were included in a further part of the study.. Tear levels of LYZ (p < 0.001), LACRT (p = 0.004) and AZGP1 (p = 0.001) were significantly elevated in GD patients with moderate-to-severe GO compared to GD patients without GO. The discriminatory power of the three biomarkers, combined in a panel was confirmed by ROC plot analysis, with an AUC value of 0.93 (sensitivity of 95%; specificity of 80%). Since LYZ showed the best performance in discriminating between GD patients with (moderate-to-severe) and without GO (in combination with limited sample volume available), LYZ levels were also measured in tears from GD patients with mild GO and without GO. Significantly higher levels of LYZ were measured in GD patients with mild GO compared to those without GO (p = 0.003).. We have established a novel three-protein biomarker panel that is able to discriminate between GD patients with and without GO, which might aid in diagnostic evaluation of GO as well as an indicator for disease activity.

Topics: Adult; Aged; Biomarkers; Diagnosis, Differential; Enzyme-Linked Immunosorbent Assay; Eye Proteins; Female; Glycoproteins; Graves Disease; Graves Ophthalmopathy; Humans; Male; Middle Aged; Muramidase; ROC Curve; Seminal Plasma Proteins; Severity of Illness Index; Tears; Young Adult; Zn-Alpha-2-Glycoprotein

To explore the differential expressions of lysozyme C and lactoferrin in tears of thyroid-associated ophthalmopathy (TAO) patients versus healthy subjects by proteomics.. Tear samples were obtained from patients with active period TAO and age and gender-matched healthy subjects without symptoms of ocular surface. Then they were divided into patient and control groups. Then tear samples of two groups were analyzed. sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of 15% gel was performed to determine the different protein bands in sample groups. And different labeled protein bands were collected for in-gel tryptic digestion. Mass spectrometry was employed to determine the protein components from different protein bands. Then Scaffold search engine was used for analyzing the results of mass spectrometry and identifying specific proteins.. Based on mass spectrometric analysis of different protein bands, most proteins were down-regulated or became absent in TAO patients. Both lysozyme C and lactoferrin were up-regulated. Identification of protein relative quantitative ratio (patient/control): lysozyme C: 4.88, lactoferrin: 1.61.. Lysozyme C and lactoferrin are two important effectors of tear function and metabolism. Both are up-regulated in TAO patients' tears. Thus both are probably involved in inflammatory process of TAO and play synergistic roles in the pathogenesis of disease.

Topics: Down-Regulation; Electrophoresis, Polyacrylamide Gel; Graves Ophthalmopathy; Humans; Lactoferrin; Mass Spectrometry; Muramidase; Proteomics; Tears; Thyroid Diseases; Up-Regulation

Proteomics and mass spectrometry are useful tools for peptide screening in body fluids. In thyroid-associated orbitopathy (TAO), evidence for lacrimal gland involvement with altered composition of tears has been reported. Our objective was to detect and evaluate potential changes in the proteomic patterns of tear fluid in TAO.. Tear fluid was collected from 45 patients with TAO and 15 healthy controls. Tear proteins were analyzed using surface-enhanced laser desorption/ionization time-of-flight mass spectrometry, and peptides were identified using matrix-assisted laser desorption/ionization time-of-flight technology.. Peptides with molecular weights 3808 Dalton (Da, p=0.004), 3734 Da (p=0.034), and 3837 Da (p=0.042), respectively, were downregulated in patients with TAO versus controls. They were identified as proline-rich protein 4 (PRP4) or as its variant nasopharyngeal carcinoma-associated PRP4. The peptide 3837 Da correlated positively with the basal secretory test (r=0.506, p<0.001) and negatively with the clinical activity score (r = -0.334, p<0.05) and age (r=-0.431, p<0.001). Also, a 12,003-Da peptide was downregulated (p=0.019) in patients and identified as ß2-microglobulin. This peptide decreased in tear fluid with increased clinical severity of TAO (p=0.027). In comparison, a 5815-Da peptide was upregulated (p=0.045) and identified as lysozyme C. When differentiating between treated and untreated patients with TAO, an 11,770-Da peptide (p=0.0072) that was also upregulated was identified as cystatin S.. Altered regulation of proinflammatory and protective proteins in tears of patients with TAO was demonstrated, reflecting an autoimmune- and/or inflammatory-induced dysfunction of the lacrimal gland.

Topics: Adult; Aged; beta 2-Microglobulin; Down-Regulation; Eye Proteins; Female; Graves Ophthalmopathy; Humans; Lacrimal Apparatus; Male; Middle Aged; Muramidase; Peptides; Proteomics; Salivary Cystatins; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Tears; Up-Regulation