Failure to demonstrate cell-mediated immunity to orbital tissue antigens and epitopic fragments of a 64 kDa protein in the majority of patients with thyroid-associated ophthalmopathy.

Abstract

We have studied a possible role of T cell sensitization to eye muscle antigens in patients with thyroid-associated ophthalmology (TAO). Peripheral blood mononuclear cell (PBMC) proliferation in response to crude porcine orbital tissue antigens, partially purified porcine eye muscle membrane proteins and predicted epitopic fragments of the recombinant 64 kDa protein 1D, was determined in patients with TAO and thyroid autoimmunity without eye disease. When membrane and cytosol fractions were used as antigen PBMC from 43% of patients with TAO but only 12.5% of normal subjects were responsive to a crude orbital connective tissue membrane fraction, although this difference was not significant. We were unable to demonstrate specific recognition of partially purified eye muscle membrane fractions; although most of the fractions tested were occasionally recognized by T cells from patients with ophthalmopathy, this was also the case for patients with autoimmune thyroid disease without ophthalmopathy and normal subjects. We did not clearly identify epitopic sequences within the 1D protein, most of the predicted peptides tested being recognized not only by T cells from a small proportion of patients with TAO, but also by those from some patients with autoimmune thyroid disease without ophthalmopathy and normal subjects. It is noteworthy however that approximately 22% of TAO patients, but no normal subjects, were positive to one or more of three peptides, suggesting that reactivity to the 1D protein may play a role in the pathogenesis of the eye disorder in some patients with TAO. The inconsistent and generally low T cell responses to crude and purified antigens noted in a few patients with TAO could be explained by low numbers of specifically sensitized lymphocytes in peripheral blood.