Immunologically mediated cytotoxicity against human eye muscle cells in Graves' ophthalmopathy

Abstract

The possible roles of antibody-mediated complement-dependent cytotoxicity (AMC), antibody-dependent killer (K) cell-mediated cytotoxicity (ADCC), and spontaneous, natural killer (NK) cell-mediated cytotoxicity (NKC) against human eye muscle cells in the pathogenesis of Graves' ophthalmopathy were investigated, using as targets human eye muscle cells, by 51Cr release assays. AMC was not demonstrated in serum from any patient or normal subject. In ADCC assays, eye muscle cell lysis was significantly increased in serum from patients with Graves' ophthalmopathy compared to those with Graves' hyperthyroidism without eye disease and normal subjects. ADCC tests were positive (percent specific lysis greater than the upper limit of normal) in 5 of 13 patients with Graves' ophthalmopathy using serum diluted 1:48 and in 4 of 10 patients using serum diluted 1:6. There was no correlation between the extent of lysis of human eye muscle and that of human (abdominal) skeletal muscle and no difference between patients with Graves' ophthalmopathy and normal subjects in assays in which abdominal muscle cell targets were used. The degree of killing in ADCC tests was independent of the source of K cells, being similar in assays using effector cells from the patient, another patient, or a normal subject. ADCC activity was partially absorbed by thyroid, orbital connective tissue and eye muscle membranes, and eye muscle cells, but not by liver membranes of thyroglobulin. Four of 8 human monoclonal antibodies reactive with eye muscle membrane antigens were cytotoxic in ADCC assays. A noncytotoxic monoclonal antibody blocked the ADCC effect of serum from a patient with Graves' ophthalmopathy, while a cytotoxic monoclonal antibody enhanced killing. NKC against eye muscle cell targets was depressed in cells from hyperthyroid and euthyroid patients with Graves' ophthalmopathy compared to that in normal subjects. Demonstration of ADCC against human eye muscle cells in some patients with Graves' ophthalmopathy suggests that this may be a mechanism for the eye muscle cell damage characteristic of this disorder. Inability to demonstrate cytotoxicity in a greater proportion of patients may reflect the lack of specific criteria to identify patients with active eye muscle inflammation and the unsuitability of currently available tests for the detection of serum antibodies against eye muscle membrane antigens. The mechanism for depressed NK cell-mediated cytotoxicity against eye muscle cells in this disorder is not known.