Endocrine Abstracts

The cellular structures lipid peroxidation related to immune system dysfunction seems to have a key role in autoimmune theory of Graves’ disease (GD). Aim: to investigate chemiluminescent (CL) activity of blood neutrophilic granulocytes in patients with GD. Materials and methods: Twenty six women, mean age 40.7±13.2, with manifestation of GD up to antithyroid therapy and 30 sex- and age-matched healthy subjects were examined. Serum thyroid hormone concentrations were measured by RIA. The median of TSH, fT4 anti-TPO concentration in GD patient were 0.17 (0.01; 0.72) mU/l, 17.9 (13.1; 30.3) pmol/l and 374.0 (223.0; 817.4) IU/ml respectively. The neutrophilic granulocytes were isolated from heparinized whole blood by double density gradient centrifugation in the ficoll-urografin: ρ=1.077 g/cm³ for lymphocytes separation, ρ=1.119 g/cm³ for the neutrophils isolation. The spontaneous and zymosan-induced (ZiCL) was studied for 90 minutes on 36-channel chemiluminescent analyzer “CL3606”. The following characteristics have been identified: time to maximum (Tmax), maximum intensity value (Imax), reflecting the maximum reactive oxygen species (ROS) level synthesis and the area under the curve (S), describing total synthesis of ROS during 90 min. of the study. Results: Whole blood CL was significantly greater in GD patients than in controls. Increased Imax of luminol-dependent spontaneous- and Zi-CL (P<0.05) of neutrophils was observed. The lucigenin-dependent CL study revealed that patients with GD have reduced the Tmax of ZiCL. In healthy controls the concentration of TSH negatively correlated with Tmax of lucigenin-dependent spontaneous(r=−0.89, P=0.019) and induced (r=−0.94; P=0.005) CL, the fT4 level is also negatively correlated with S (r=−0.83, P=0.042) of lucigenin-dependent spontaneous CL, Tmax (r=−0.83, P=0.042) and S (r=−0.94; P=0.005) of lucigenin-dependent induced CL of neutrophils. In GD patients interaction in pairs of anti-TPO/Tmax (r=−0.70, P=0.036) and anti-TPO/Imax(r=−0.72, P=0.030) of luminol-dependent induced CL was observed. Conclusion. Hyperactivity and reactivity of peripheral blood neutrophils in GD certify by respiratory explosion in spontaneous and induced of secondary ROS level synthesis. Enhanced excitability to phagocytosis-related stimuli can mediate the autoimmune mechanism in GD.

2. The relationship between indicators of respiratory burst of neutrophils and thyroid hormone when BG is lost, but there is a moderate negative correlation between CL response and the level of at-TPO.

Keywords: graves disease, neutrophilic granulocyte, chemiluminescence, reactive oxygen species