Zmiana ekspresji mRNA dla CTLA-4, CD28, VDR i CD45 w limfocytach T u osób z chorobą Hashimoto — badanie pilotowe

Introduction: CD28/T-cell receptor (TCR)/cytotoxic T-lymphocyte antigen 4 (CTLA4) complex controls T-cell tolerance and autoimmunity in Hashimoto’s thyroiditis (HT). In addition, CD45 protein tyrosine phosphatase (PTPase) and vitamin D receptor (VDR) cooperatively interact with the TCR complex to affect autoimmune processes central to the pathogenesis of HT. Nevertheless, their role in HT aetiology has been less well established. In this study, we aimed to explore mRNA expression levels of CTLA4, CD28, CD45, and VDR in T-cells, across different outcomes of HT. Material and methods: The study included 45 HT patients and 13 euthyroid, healthy controls. T-lymphocytes were isolated from peripheral blood mononuclear cells, total mRNA was extracted from T-cells, and gene expression was studied by reverse transcription-polymerase chain reaction (RT-PCR) and ImageQuant method relative to glyceraldehyde-3-phosphate dehydrogenase RT-PCR products. Results: Nominally higher expression levels of VDR, CTLA4, CD28, and CD45RAB mRNA were found in unsorted T-lymphocytes of healthy controls when compared to the HT patients. No difference was observed between hypothyroid/untreated, spontaneously euthyroid and LT4-treated HT patients. VDR mRNA expression was linked to both T3 levels and CTLA4 gene expression, whilst CD45RB mRNA expression coincided with CTLA4 and CD28 transcript levels. Conversely, older age and lower T3 levels were associated with increased abundance of CD45R0 isoform in HT patients. Conclusions: The results suggest a cross talk between endocrine and immune functions in HT pathology: an altered peripheral T cell mRNA profile with reduced VDR, CTLA4, CD28, and CD45RAB transcript levels is accompanied by age-related shift from naive to memory/late-differentiated T cell CD45R mRNA signature and associated with thyroid hormone status in the HT patients.Wstęp: Kompleks antygenu CD28/receptora limfocytów T (TCR)/antygenu 4 związan ego z limfocytem T cytotoksycznym (CTLA4) reguluje tolerancję limfocytów T oraz autoimmunogenność w chorobie Hashimoto (HT). Ponadto białkowa fosfataza tyrozynowa (PTPase) CD45 oraz receptor witaminy D (VDR) wchodzą w interakcję z kompleksem TCR, modyfikując procesy autoimmunologiczne mające podstawowe znaczenie w patogenezie HT. Jednak rola tych cząsteczek w etiologii HT nie została dokładnie ustalona. Celem autorów badania była ocean poziomów ekspresji mRNA dla CTLA4, CD28, CD45 i VDR w limfocytach T w zależności od różnego statusu HT. Materiał i metody: Do badania włączono 45 chorych na HT i 13 zdrowych osób z prawidłową czynnością tarczycy. Limfocyty T wyizolowano spośród komórek jędnojądrzastych krwi obwodowej, wyekstrahowano z nich całkowity mRNA i określono ekspresję genów za pomocą łańcuchowej reakcji polimerazowej z odwrotną transkryptazą (RT-PCR) i metody ImageQuant związanej produktami reakcji RT-PCR dehydrogenazy aldehydu 3-fosfoglicerynowego. Wyniki: U osób zdrowych stwierdzono nominalnie wyższy poziom ekspresji mRNA dla VDR, CTLA4, CD28 i CD45RAB w niesortowanych limfocytach T niż u chorych na HT. Nie zaobserwowano żadnych różnic między chorymi na HT z niedoczynnością tarczycy/nieleczonymi, u których samoistnie nastąpiło przywrócenie eutyreozy, i stosującymi leczenie LT4. Ekspresja VDR mRNA była powiązana zarówno ze stężeniem T3, jak i ekspresją genu CTLA4, natomiast ekspresja mRNA dla CD45RB wiązała się z poziomami transkryptu CTLA4 i CD28. Z kolei starszy wiek i niższe stężenia T3 wiązały się ze zwiększoną ilością izoformy CD45R0 u chorych na HT. Wnioski: Uzyskane wyniki sugerują interferencje między czynnością wewnątrzwydzielniczą i immunologiczną w patologii HT: zmiana profilu mRNA obwodowych limfocytów T z ograniczeniem poziomu transkryptu białek VDR, CTLA4, CD28 i CD45RAB współistnieje z zależnym od wieku przesunięciem sygnatury mRNA limfocytów CD45R od komórek naiwnych do komórek pamięci immunologicznej/ zróżnicowanych i jest powiązana ze stężeniami hormonów tarczycy u chorych na HT

PAX8-PPARg Oncogene in Follicular Thyroid Tumors: RT-PCR and Immunohistochemical Analyses

US-guided fine needle aspiration cytology is currently the best diagnostic tool for thyroid nodules. However, it is not sensitive and specific enough for differentiating between benign and malignant follicular tumors. A potentially useful marker for this differentiation is the PAX8-PPARg rearrangement, identified in follicular thyroid carcinomas, but not in follicular adenomas or other types of thyroid tumors. The aim of this research was to determine the clinical significance of the PAX8-PPARg oncogene in diagnostics follicular thyroid tumors. The study included 62 patients with follicular or Hürthle cell tumors. Gene expression was determined by reverse transcription-polymerase chain reaction (RT-PCR) from paraffin embedded tissues, and PCR products were checked using the agarose gel electrophoresis. The immunohistochemical analysis was performed on archive paraffin embedded tissues with the monoclonal PPARã antibody. The statistical analysis has indicated that neither the expression of PAX8-PPARg mRNA, nor the immunohystochemical analysis with the PPARg antibody correlate with the patohystological diagnosis. The oncogene PAX8-PPARg has not met the expectations as a reliable tumor marker for differentiation between benign and malignant thyroid tumors, which makes the only reliable histological criteria – capsular and vascular invasion

PAX8-PPARg Oncogene in Follicular Thyroid Tumors: RT-PCR and Immunohistochemical Analyses

US-guided fine needle aspiration cytology is currently the best diagnostic tool for thyroid nodules. However, it is not sensitive and specific enough for differentiating between benign and malignant follicular tumors. A potentially useful marker for this differentiation is the PAX8-PPARg rearrangement, identified in follicular thyroid carcinomas, but not in follicular adenomas or other types of thyroid tumors. The aim of this research was to determine the clinical significance of the PAX8-PPARg oncogene in diagnostics follicular thyroid tumors. The study included 62 patients with follicular or Hürthle cell tumors. Gene expression was determined by reverse transcription-polymerase chain reaction (RT-PCR) from paraffin embedded tissues, and PCR products were checked using the agarose gel electrophoresis. The immunohistochemical analysis was performed on archive paraffin embedded tissues with the monoclonal PPARã antibody. The statistical analysis has indicated that neither the expression of PAX8-PPARg mRNA, nor the immunohystochemical analysis with the PPARg antibody correlate with the patohystological diagnosis. The oncogene PAX8-PPARg has not met the expectations as a reliable tumor marker for differentiation between benign and malignant thyroid tumors, which makes the only reliable histological criteria – capsular and vascular invasion

The effects of suppressive therapy of difuse euthyroid goitre with levothyroxine (comparation on effects in younger and middle-aged patients)

U prospektivnoj studiji kojom je obuhvaćeno 67 bolesnika s difuznom eutireoidnom strumom, starosne dobi od 17 do 21 god. (grupa I n=28) i dobi od 33 do 45 god. (grupa II, n=39), liječenih godinu dana sa 100 ug tiroksina dnevno, ocijenjeni su učinci supresijske terapije ultrazvučnom volumetrijom i palpacijskim nalazom. Nakon 3, 6, 9 i 12 mjeseci praćenja uslijedilo je značajno smanjenje volumena strume u obje grupe bolesnika u odnosu na početnu veličinu strume. U obje grupe volumen struma najviše se smanjio u prvih 6 mjeseci primjene terapije. Poslije 6 mjeseci prosječno smanjenje volumena struma u grupi I iznosilo je 19.2%, u grupi II 28.6%, a nakon 12 mjeseci u grupi I volumen struma smanjen je za 23.5%, a u grupi II za 32.3% u odnosu na početni volumen. U grupi I nakon mjesec dana (u grupi II nakon dva mjeseca) uočen je značajan porast tiroksina (T4) i njegove slobodne frakcije (fT4) u odnosu na normalne vrijednosti prije početka primjene supresijske terapije, dok se vrijednosti trijodtironina (T3) i njegove slobodne frakcije (fT3) nisu mijenjale. U obje grupe u prvih 6 mjeseci i nakon godinu dana terapije tiroksinom ustanovljena je učinkovitost u smanjenju strume sa statistički signifikantnom razlikom u korist grupe II.In a prospective study 67 patients with diffuse euthyroid goitre, between 17 and 21 years old (group I, n=28) and between 33 and 45 years old (group II, n=39), were treated for one year with 100 levothyroxine daily, have been evaluated effects of supressive therapy measured by sonography and palpation. After 3, 6, 9 and 12 months the reduction of the goitre volume was significantly different from the initial volume in both groups. In both forms of treatment maximum volume decrease was already seen within the first six months. After treatment for 6 months the mean decrease of thyroid volumes was 19.2% in group I and 28.6% in group II. After treatment for 12 months the mean decrease of thyroid volumes was 23.5% in group I and 32.3% in group II compared to the initial volume. In group I after 1 month (in group II after 2 months) a more significant increase of T4 and of fT4 values within the normal range was observed immediately after supressive therapy. There were no changes of the T3 and fT3 values. In both groups it has been found out that in the first 6 months and after 1 year therapy with levothyroxine exist a comparable efficiency for the reduction of goiters with statistically significant differences in favour group II

Thyroid Autoimmunity and Infertility

Autoimmune thyroid disease (AITD) is one of the most common endocrinopathies and is more prevalent in women. The circulating thyroid antibodies and the hypothyroidism that often follows AITD have effects on many tissues. The endometrium and ovaries are not spared, and therefore this common morbidity might have an impact on fertility. Despite challenging data interpretation and contradictory results, a general takeaway from published studies is that there is a higher incidence of elevated levels of thyroid-stimulating hormone (TSH) and the presence of thyroid antibodies among infertile women. While a single specific and direct pathophysiological mechanism through which autoimmune thyroid disease causes infertility has not been identified, there are multiple gynecological comorbidites that might perpetuate infertility (endometriosis, premature ovarian failure, polycystic ovaries) and defective immunological functions (a shift to a proinflammatory Th1 response, increased levels of natural killer cells, cross-reactivity of antigens etc.) that are affecting fertility. There is insufficient evidence that suggests levothyroxine (LT4) treatment can help women suffering from AITD conceive and carry out a pregnancy

The effects of suppressive therapy of difuse euthyroid goitre with levothyroxine (comparation on effects in younger and middle-aged patients)

U prospektivnoj studiji kojom je obuhvaćeno 67 bolesnika s difuznom eutireoidnom strumom, starosne dobi od 17 do 21 god. (grupa I n=28) i dobi od 33 do 45 god. (grupa II, n=39), liječenih godinu dana sa 100 ug tiroksina dnevno, ocijenjeni su učinci supresijske terapije ultrazvučnom volumetrijom i palpacijskim nalazom. Nakon 3, 6, 9 i 12 mjeseci praćenja uslijedilo je značajno smanjenje volumena strume u obje grupe bolesnika u odnosu na početnu veličinu strume. U obje grupe volumen struma najviše se smanjio u prvih 6 mjeseci primjene terapije. Poslije 6 mjeseci prosječno smanjenje volumena struma u grupi I iznosilo je 19.2%, u grupi II 28.6%, a nakon 12 mjeseci u grupi I volumen struma smanjen je za 23.5%, a u grupi II za 32.3% u odnosu na početni volumen. U grupi I nakon mjesec dana (u grupi II nakon dva mjeseca) uočen je značajan porast tiroksina (T4) i njegove slobodne frakcije (fT4) u odnosu na normalne vrijednosti prije početka primjene supresijske terapije, dok se vrijednosti trijodtironina (T3) i njegove slobodne frakcije (fT3) nisu mijenjale. U obje grupe u prvih 6 mjeseci i nakon godinu dana terapije tiroksinom ustanovljena je učinkovitost u smanjenju strume sa statistički signifikantnom razlikom u korist grupe II.In a prospective study 67 patients with diffuse euthyroid goitre, between 17 and 21 years old (group I, n=28) and between 33 and 45 years old (group II, n=39), were treated for one year with 100 levothyroxine daily, have been evaluated effects of supressive therapy measured by sonography and palpation. After 3, 6, 9 and 12 months the reduction of the goitre volume was significantly different from the initial volume in both groups. In both forms of treatment maximum volume decrease was already seen within the first six months. After treatment for 6 months the mean decrease of thyroid volumes was 19.2% in group I and 28.6% in group II. After treatment for 12 months the mean decrease of thyroid volumes was 23.5% in group I and 32.3% in group II compared to the initial volume. In group I after 1 month (in group II after 2 months) a more significant increase of T4 and of fT4 values within the normal range was observed immediately after supressive therapy. There were no changes of the T3 and fT3 values. In both groups it has been found out that in the first 6 months and after 1 year therapy with levothyroxine exist a comparable efficiency for the reduction of goiters with statistically significant differences in favour group II