Expression of genes for certain enzymes of pyrimidine and purine salvage pathway in peripheral blood leukocytes collected from patients with Graves' or Hashimoto's disease

Abstract

Increased activities of some enzymes, which participate in pyrimidine and purine salvage pathway, were found in blood fractions of patients suffering from different autoimmunological diseases, thyroid diseases included. The aim of the study was to estimate the expression of genes, specific for deoxycytidine kinase (dCK, EC 3.7.1.74), thymidine kinase 1 (TK1; EC 2.7.1.21), and adenosine deaminase (ADA, EC 3.5.4.4) in blood leukocytes, collected from patients with autoimmunological thyroid diseases (AITD), i.e., Graves' or Hashimoto's disease. The total mRNA was isolated from peripheral blood leukocytes and, afterwards, submitted to reverse transcription (RT), with the following amplification of genes encoding for particular examined enzymes and beta-actin, as a supervisory gene [RT-polymerase chain reaction (RT-PCR)]; ADA gene was amplified with the use of three different primer pairs (ADA3, ADA4, and ADA5). PCR products were electrophoresed in 8% polyacrylamide gel and then, submitted to densitometric analysis. The levels of expression of all the examined genes in leukocytes from patients with either Graves' or Hashimoto's disease were significantly increased when compared to those in controls; above a twofold elevation of expression of TK1, ADA4, and ADA5 genes was observed. In conclusion, the changes of activities of salvage enzymes in patients with AITD occur likely at transcription level; the measurement of gene expression for purine and pyrimidyne salvage enzymes may likely help explain the mechanism of autoimmune diseases, being also significant in the diagnostics and/or monitoring of AITD.

The association between serum adenosine deaminase levels and Graves’ disease

“…In some autoimmune diseases, such as psoriasis ( 7 ), autoimmune hepatitis ( 8 ), inflammatory bowel disease ( 9 ), and rheumatic disease ( 10 ), high serum concentrations of ADA were observed. Meanwhile, increased ADA activity was also observed in the peripheral blood leucocytes of patients with GD ( 11 ) and monocytes of patients with Hashimoto disease ( 12 ). Therefore, serum ADA level may have the potential to be an indicator for the evaluation and monitoring of GD.…”

Moonlighting Adenosine Deaminase: A Target Protein for Drug Development

“…Changes in normal levels of ADA have been reported in a wide variety of pathologies . For example, alterations in the amount of one or both isoenzymes have been observed in a number of diseases such as hereditary hemolytic and Diamond Blackfan anemias, tuberculosis, systemic lupus erythematosus, inflammatory responses, rheumatoid arthritis, heart diseases, acquired immunodeficiency syndrome, autoimmunological thyroid diseases, some types of carcinoma, and neurological disorders, such as multiple sclerosis, meningitis, fibromyalgia, depression, and panic disorder …”

Clinical biomarkers for thyroid immune-related adverse events in patients with stage III and IV gastrointestinal tumors

“…Our investigation identified a higher baseline level of ADA (>14.05 U/L) as an independent biomarker associated with thyroid irAEs. ADA, a key enzyme involved in purine metabolism and immune system regulation, has been associated with thyroid autoimmune diseases at the transcriptional level ( 14 ). It can activate T lymphocytes ( 15 ) as well as alleviate immunosuppressive effects by degrading adenosine ( 16 ).…”