Rola infekcji wirusem Epstein-Barr’a w rozwoju autoimmunologicznych chorób tarczycy

Introduction: Autoimmune thyroid diseases, including Graves’ and Hashimoto’s thyroiditis, are the most frequent autoimmune disorders. Viral infection, including Epstein-Barr virus (EBV), is one of the most frequently considered environmental factors involved in autoimmunity. Its role in the development of AITD has not been confirmed so far.Material and methods: Surgical specimens of Graves’ and Hashimoto’s diseases and nodular goitres were included in the study. The expression of EBV latent membrane protein 1 (LMP1) was analysed by immunohistochemistry, with the parallel detection of virus-encoded small nuclear non-polyadenylated RNAs (EBER) by in situ hybridisation.Results: In none of the Graves’ disease specimens but in 34.5% of Hashimoto’s thyroiditis cases the cytoplasmic expression of LMP1 was detected in follicular epithelial cells and in infiltrating lymphocytes. EBER nuclear expression was detected in 80.7% of Hashimoto’s thyroiditis cases and 62.5% of Graves´ disease cases, with positive correlation between LMP1 and EBER positivity in all Hashimoto’s thyroiditis LMP1-positive cases.Conclusions: We assume that high prevalence of EBV infection in cases of Hashimoto’s and Graves’ diseases imply a potential aetiological role of EBV in autoimmune thyroiditis. The initiation of autoimmune thyroiditis could start with EBV latency type III infection of follicular epithelium characterised by LMP1 expression involving the production of inflammatory mediators leading to recruitment of lymphocytes. The EBV positivity of the infiltrating lymphocytes could be only the presentation of a carrier state, but in cases with EBER+/ LMP1+ lymphocytes (transforming latent infection) it could represent a negative prognostic marker pointing to a higher risk of primary thyroid lymphoma development. (Endokrynol Pol 2015; 66 (2): 132–136)Wstęp: Autoimmunologiczne choroby taczycy, w tym zapalenia tarczycy Graves’a i Hashimoto, są najczęstszymi zaburzeniami autoimmunologicznymi. Infekcja wirusowa, w tym wirusem Epstein-Barr’a (EBV), jest jednym z najczęściej rozważanych czynników środowiskowych łączonych z autoimmunologią. Jego rola w rozwoju autoimmunologicznyh chorób taczycy (AITD) nie została do tej pory potwierdzona.Materiały i metody: Zbadano chirurgicznie pobrane preparaty od chorych na chorobę Gravesa, chorobę Hashimoto i wole guzowate. Ekspresja latentnego białka błonowego 1 (LMP1) EBV była zanalizowana metodami immunohistochemicznymi, z równoczesnym wykrywaniem kodowanych wirusem, małych, jądrzastych, niespoliadenylowanych RNA (EBER) poprzez hybrydazcję in situ.Wyniki: W żadnym z przypadków choroby Gravesa nie stwierdzono w komórkach pęcherzykowych i naciekających limfocytach ekspresji LMP1w cytoplazmie, ale wykryto ją w 34,5% przypadkach choroby Hashimoto. Jądrowa ekspresja EBER została wykryta w 80,7% przypadków zapalenia tarczycy Hashimoto i w 62,5% choroby Gravesa. Zauważono również dodatnią korelację pomiędzy LMP1 i występowaniem EBER we wszystkich przypadkach choroby Hashimoto LMP1+.Wnioski: Autorzy uwazają, że powszechne występowanie infekcji EBV w chorobach Hashimoto i Gravesa sugeruje potencjalną rolą etiologiczną EBV w rozwoju autoimmunologicznych zapaleń tarczycy. Zapoczątkowaniem autoimmunologicznych zapaleń tarczycy może być utajona infekcja EBV typu III, charakteryzująca się ekspresją LMP1 w komórkach pęcherzykowych i związana z produkcją mediatorów zapalnych prowadzącą do migracji limfocytów. Obecność EBV w naciekach limfocytarnych może być jedynie charakterystyczna dla nosiciela wirusa ale w przypadku obecności limfocytów EBER+/LMP1+ (przekształcajcych infekcję utajoną) może być negatywnym markerem prognostycznym wskazując podwyższone ryzyko rozwoju pierwotnego chłoniaka tarczycy. (Endokrynol Pol 2015; 66 (2): 132–136

Report of three novel germline CYLD mutations in unrelated patients with Brooke-Spiegler syndrome, including classic phenotype, multiple familial trichoepitheliomas and malignant transformation

Brooke-Spiegler syndrome is a rare autosomal-dominant genetic disorder characterized by multiple adnexal tumors, including cylindromas, spiradenomas, spiradenocylindromas and trichoepitheliomas. It is caused by germline CYLD mutations commonly leading to a premature stop codon. We here report on 3 novel CYLD mutations in 3 unrelated BSS patients, including the classic phenotype, multiple familial trichoepitheliomas phenotype and malignant transformation. These included c.1821_1826+1delinsCT/L607Ffs*9, c.2666A>T/p.D889V and c.2712delT/p.905Kfs*8. By extending the spectrum of CYLD mutations, better understanding of the molecular mechanisms of BSS can be gained, which might later assist in finding new treatment options

Gene-Directed Enzyme/Prodrug Therapy of Rat Brain Tumor Mediated by Human Mesenchymal Stem Cell Suicide Gene Extracellular Vesicles In Vitro and In Vivo

MSC-driven, gene-directed enzyme prodrug therapy (GDEPT) mediated by extracellular vesicles (EV) represents a new paradigm—cell-free GDEPT tumor therapy. In this study, we tested the efficacy of yeast cytosine deaminase::uracilphosphoribosyl transferase (yCD::UPRT-MSC)-exosomes, in the form of conditioned medium (CM) to inhibit the growth of C6 glioblastoma cells both in vitro and in vivo. MSCs isolated from human adipose tissue, umbilical cord, or dental pulp engineered to express the yCD::UPRT gene secreted yCD::UPRT-MSC-exosomes that in the presence of the prodrug 5-fluorocytosine (5-FC), inhibited the growth of rat C6 glioblastoma cells and human primary glioblastoma cells in vitro in a dose-dependent manner. CM from these cells injected repeatedly either intraperitoneally (i.p.) or subcutaneously (s.c.), applied intranasally (i.n.), or infused continuously by an ALZET osmotic pump, inhibited the growth of cerebral C6 glioblastomas in rats. A significant number of rats were cured when CM containing yCD::UPRT-MSC-exosomes conjugated with 5-FC was repeatedly injected i.p. or applied i.n. Cured rats were subsequently resistant to challenges with higher doses of C6 cells. Our data have shown that cell-free GDEPT tumor therapy mediated by the yCD::UPRT-MSC suicide gene EVs for high-grade glioblastomas represents a safer and more practical approach that is worthy of further investigation

HLA-G 14bp Ins/Del Polymorphism, Plasma Level of Soluble HLA-G, and Association with IL-6/IL-10 Ratio and Survival of Glioma Patients

HLA-G is an immune checkpoint molecule with immunosuppressive and anti-inflammatory activities, and its expression and level of its soluble form (sHLA-G) may play an important role in tumor prognosis. The HLA-G 14bp ins/del polymorphism and the plasma level of soluble HLA-G (sHLA-G) were investigated by a polymerase chain reaction and ELISA, respectively, in 59 glioma patients. A significantly higher proportion of glioma patients had the 14 nt insert in both homozygous and heterozygous states compared to the control group. Glioma patients also had higher plasma levels of sHLA-G. Patients with methylated MGMT promoters had lower levels of sHLA-G than those with unmethylated MGMT promoters. The level of sHLA-G negatively correlated with the overall survival of patients. Glioblastoma patients who survived more than one year after diagnosis had lower levels of sHLA-G than those surviving less than one year. Patients with sHLA-G levels below the cut-off value of 40 U/mL survived significantly longer than patients with sHLA-G levels above 40 U/mL. The levels of sHLA-G were also negatively correlated with the level of IL-6 (p = 0.0004) and positively with IL-10/IL-6 (p = 0.046). Conclusion: The presence of the 14 nt insert in both homozygous and heterozygous states of the HLA-G 14bp ins/del polymorphism is more frequent in glioma patients and the elevated plasma levels of sHLA-G are negatively associated with their survival