Different Cytokine mRNA Profiles in Graves' Disease, Hashimoto's Thyroiditis, and Nonautoimmune Thyroid Disorders Determined by Quantitative Reverse Transcriptase Polymerase Chain Reaction (RT-PCR)

Abstract

Intrathyroidal lymphocytes are a source of cytokines thought to stimulate or maintain the immune process within the thyroid in Graves' disease (GD) and Hashimoto's thyroiditis (HT). Quantitative assessment of the cytokine profile may provide important clues as to the Th1/Th2 balance prevailing in these diseases. We analyzed cytokine mRNA expression levels in thyroid tissue samples from 13 patients with GD, 2 with HT, 5 with nontoxic multinodular goiter (NTG), and 4 with thyroid autonomy (nodular = TAnod and perinodular = TAperi tissue) using multispecific competitor fragments with primer sequences for IL-1 beta, IL-2, IL-4, IL-6, IL-8, IL-10, IFN-gamma, CD25, and CD3 delta-chain mRNA. Patients with GD were subdivided into two groups according to their serum levels of antibodies to thyroperoxidase (anti-TPO; GDhigh > 4000 U/mL, GDlow < or = 200 U/mL). These levels correlated positively with the CD3 delta-chain mRNA levels (r = 0.83) and with the T cell infiltration (r = 0.71) as determined by immunohistochemistry. Patients with GDhigh demonstrated 2- to 4-fold higher IL-4 mRNA levels (as compared to all other investigated groups) and significantly higher IL-10 mRNA levels as compared to HT, GDlow, and TAnod patients. Patients with GDhigh also had significantly higher levels of IFN-gamma, IL-1 beta, IL-8, and CD25 mRNA as compared to GDlow. The highest IFN-gamma, IL-2, and CD25 mRNA levels were found in HT. The lowest mRNA levels of all the investigated groups were detected in TAnod. No significant differences in IL-6 and IL-8 mRNA levels were found between most of the patient groups. In summary, patients with GDhigh showed a shift to a more Th2-driven cytokine pattern. In contrast, the increase mRNA levels of Th1-related cytokines found in HT indicate predominantly T cell-mediated cytotoxic processes.

Iodine-131 therapy alters the immune/inflammatory responses in the thyroids of patients with Graves' disease

“…In patients with GD, IL-6 is produced by infiltrating immune cells and thyrocytes (25). Serum IL-6 levels in patients with GD have been reported to be significantly higher than those in healthy controls (21,26). However, the clinical significance of serum IL-6 levels remains controversial.…”

“…Wahrenberg et al (21) have determined that the serum level of IL-6 indicates the tendency of inflammatory response. Furthermore, Heuer et al (26) claim that the elevated IL-6 concentration may reflect disease severity. In the present study, it was demonstrated that serum IL-6 levels in patients with GD were significantly higher than those in the control group.…”

The effects of CD40- and interleukin (IL-4)-activated CD23+ cells on the production of IL-10 by mononuclear cells in Graves’ disease: the role of CD8+ cells

“…There was no difference between the high and low basal secretion groups as far as clinical findings are concerned. Heuer et al [29] reported that patients with Graves' disease could be subdivided into two groups according to their serum levels of anti-TPO antibodies. Patients with an anti-TPO antibody titre of more than 4000 U/ml showed greater T cell infiltration and higher levels of IL-4 and IL-10 mRNA in their thyroid tissues compared to those with antibody of less than 200 U/ml.…”

Expression of tumour necrosis factor-alpha (TNF-α) mRNA and protein in pathological thyroid tissue and carcinoma cell lines

“…0046-4 . 6 fg/l) to the RT-PCR reaction mixtures containing aliquots of cDNA according to the method of Platzer et al [16] and described in detail by Heuer et al [17]. Since the CF and the target cDNA were amplified with the same primers (Table 1), amplification products obtained from the CF are shorter in size than the target cDNA products.…”