Detection of interleukin-6 and interleukin-1 production in human thyroid epithelial cells by non-radioactivein situhybridization and immunohistochemical methods

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Abstract
Human endocrine thyroid epithelial cells have been described to produce cytokines in vitro. In order to determine whether they do so in vivo during thyroiditis. parallel studies on mRNA expression with a non-radioactive in situ hybridization technique and immunohislochemical detection for the protein were performed on frozen sections of thyroid samples from autoimmune thyroiditis (Graves' disease and Hashimoto's thyroiditis), non-toxic goitre and normal thyroid tissue. cDNA probes were sulphonated and their hybridization with mRN A was detected with a sulphonyl-specific monoclonal antibody. This signal was amplified and visualized with the alkaline phosphatase anti-alkaline phosphatase (APAAP) system. The protein products were detected with immuno-purified rabbit F(ab′)2 antibody fragments recognizing recombinant human cytokines, visualized by the immuno-peroxidase technique. Each sample was studied at the two levels. Both interleukin-6 mRNA and protein were found in the endocrine cells. There was no obvious difference between autoimmune thyroiditis and non-toxic goitre. However, normal thyroid epithelial cells produced less interleukin-6. Interleukin-1α mRNA and its protein were found in epithelial cells from Hashimoto's thyroiditis samples, but not in the others, except one Graves' disease sample, in which only mRNA was detected. Interleukin- 1β was not detected in these cells, its mRNA was only found in one of the Graves' disease samples. These cytokines were also detected in some infiltrating cells.