O,O'-diethyl-(S,S)-ethylenediamine-N,N'-di-2-(3-cyclohexyl)propanoate dihydrochloride enhances influx of effective NK and NKT cells in murine breast cancer

Background/Aim. O,O'-diethyl-(S,S)-ethylenediamineN,N'-di-2-(3-cyclohexyl)propanoate dihydrochloride (DE-EDCP) has been found to possess promising cytotoxic activity against various tumor cell lines. Also, DE-EDCP reduces tumor progression by several mechanisms such as triggering tumor cell death and inhibition of cell proliferation. The aim of present study was to further evaluate antitumor activity of DE-EDCP by investigating effects on migratory potential of tumor cells and anti-tumor immune response. Methods. Migratory potential of DE-EDCP was evaluated by scratch wound assay. Female BALB/c mice were inoculated with 4T1 breast cancer cells and treatment with DE-EDCP started five days following orthotopic tumor implantation. The frequency and phenotype of tumor-infiltrating natural killer (NK) and natural killer T (NKT) cells were analyzed by flow cytometry. Results. DE-EDCP inhibited migratory potential of highly metastatic 4T1 cells. DE-EDCP facilitated accumulation of CD3+CD49+ NKT cells and CD3-CD49+ NK cells in tumor microenvironment. DE-EDCP treatment led to significant decrement of tumor infiltrating anergic NKT cells expressing cytotoxic T-lymphocyte-associated protein 4 (CTLA-4), killer cell lectin like receptor G1 (KLRG-1) and programmed cell death protein-1 (PD-1). Mice given DE-EDCP had significantly increased percentages of tumoricidal fas ligand (FasL) positive NK cells. Conclusion. DE-EDCP inhibits murine breast cancer progression through direct effects on tumor cells and by facilitating anti-tumor immunity. DE-EDCP enhances accumulation, promotes tumoricidal phenotype and maintenances responsiveness of NK and NKT cells in 4T1 murine breast cancer

Auto imunska bolest štitaste žlezde - patogeneza Graves-ove bolesti i Hashimoto tireoiditisa

It is generally accepted that autoimmune thyroid disorders, Graves disease and Hashimoto thyroiditis, differ in pathogenesis and clinical implications. In Graves disease the basic pathogenetic mechanism is B lymphocyte activation which produce auto antibodies specific for TSH receptor (TSHR) which binding to thyrocyte membrane causes their long-termed stimulation which gives as a result the occurrence of hyperthyrosis. On the other hand in Hashimoto thyroiditis lymphocyte accumulation occurs and they cause gradual thyrocyte damage and hypothyrosis development. However, it was found that the reisa certain genetic predisposition for both autoimmune thyroid diseases and that they can appear among several members of the same family. Besides in the serum of the patients with Graves disease and Hashimoto thyroiditis the presence of autoantibodi esspecific for dominant thyroid autoantigenes: TSHR, thyroperoxidase (TPO) and thyroglobulin (Tg) can be found as indicators of the auto immune process in thyroid gland. For these reasons both autoimmune diseases of thyroid gland sometimes are marked with the common name: autoimmune thyroid disease. As cell and molecule mechanisms included in initiation of the autoimmune process in thyroid gland that define the type and natural course of disease have not completely been explained, in this review the data from the literature considering pathogenesis of autoimmune diseases of thyroid gland have been shown. After introductory considerations, dominant autoantigenes and autoantibodies (as indicators of autoimmune process in thyroid gland) are shown in details, as well as mechanisms included in effector phase of autoimmune process which cau se the thyroid cell damage. The role of disturbance in regulation of the apoptosis process is especially analyzed as they could effect the development of autoimmune diseases of thyroid gland.Prihvaćeno je shvatanje da se autoimunske bolesti štitaste žlezde Graves-ova bolest i Hashimoto tireoiditis razlikuju po patogenezi i kliničkim posledicama. U Graves-ovoj bolesti je osnovni patogenetski mehanizam aktivacija Blimfocita koji produkuju auto antitela specifična za TSH receptor (TSHR), čije vezivanje za membranu tireocita uzrokuje njihovu dugotrajnu stimulaciju, sa posledičnim nastankom hipertireoze. S druge strane, u Hashimoto tireoiditi su nastaje akumulacija limfocita koji prouzrokuju postepeno oštećenje tireocita i nastanak hipotireoze. Međutim utvrđeno je da za obe autoimunske tireoidne bolesti postoji određena genetska predispozicija i da se one mogu javiti kod više članova u istoj porodici. Osim toga, u serumu obolelih od Graves-ove bolesti i Hashimoto tireoiditi sa se može pokazati prisustvo autoantitela specifičnih za dominantne tireoidne autoantigene (receptor za TSH, tireoperoksidazu i tireoglobulin) koji predstavljaju pokazatelje autoimunskog procesa u štitastoj žlezdi. Iz tih razloga se obe autoimunske bolesti štitaste žlezde nekad označavaju zajedničkim nazivom: autoimunska bolest štitaste žlezde. Budući da ćelijski i molekulski mehanizmi koji su uključeni u inicijaciju autoimunskog procesa u štita stoj žlezdi, i opredeljuju vrstu i prirodni tok bolesti, nisu potpuno rasvetljeni, u ovom radu su prikazani podaci iz literature koji se odnose na patogenezu autoimunskih bolesti štitaste žlezde. Nakon uvodnih razmatranja, detaljno su prikazani dominantni autoantigeni i autoantitela (kao pokazatelji autoimunskog procesa u štitastoj žlezdi), kao i mehanizmi uključeni u efektorsku fazu autoimunskog procesa koji uzrokuju oštećenje tireocita. Posebno je analizirana uloga poremećaja u regulaciji procesa apoptoze u nastanku autoimunskih bolesti štitaste žlezde

Latent Murine Cytomegalovirus Infection Contributes to EAE Pathogenesis

ABSTRACT Viral infection has been identified as the most likely environmental trigger of multiple sclerosis (MS). There are conflicting data regarding the role of cytomegalovirus (CMV) in MS pathogenesis. We utilised experimental autoimmune encephalomyelitis (EAE)-resistant BALB/c mice and murine cytomegalovirus (MCMV), the murine homolog of CMV, to examine the mechanism by which viral infection enhances autoimmune neuroinflammation. Mice subjected to latent neonatal MCMV infection developed the typical characteristics of EAE. Similar to MS, the MCMV-infected EAE-induced mice developed infiltrates in the central nervous system (CNS) composed of similar percentages of CD4+ and CD8+ T cells. The influx of both Th 1 and Th 17 cells into the CNS of MCMV- infected EAE-induced mice was observed. Interestingly, the development of autoimmune neuroinflammation after latent MCMV infection was accompanied by a significant influx of Tc17 cells (CD8+IL-17+ and CD8+RoRγt+) but not Tc1, cells. Our results suggest that latent MCMV infection affects the development of inflammatory lymphocytes that exhibit encephalitogenic potential, thereby mediating increased CNS pathology following EAE induction, and that CMV represents a possible environmental factor in the pathogenesis of MS and other autoimmune disease

Korelacija koncentracija autoantitela specifičnih za tiroidnu peroksidazu određenih korišćenjem dva radioimunološka testa

Introduction. Thyroid peroxidase-specific autoantibodies (TPO Abs) are mostly measured in patients with autoimmune thyroid diseases. The aim of this study was to compare TPO Ab concentrations measured by two radioimmunoassays. Material and methods. Our investigation included 38 patients. Sera concentrations of TPO Abs were measured by using Cis biointernational (France) and Immunotech (Czech Republic) assays. Results. Concentrations obtained by two assays were extensively different. The values measured by Cis biointernational assay were higher than ones obtained by Immunotech assay. The statistical arrangement of results showed the direct correlation between the two assays, with the coefficient of agreement R=0.6239 (p lt 0.001). The analysis of relative values (ratio of measured and upper limit values given by the manufacturer) demonstrated the statistically significant difference (p=0.003) between values measured by Cis biointernational (18.94±37.22) and by Immunotech assay (4.22±8.22) concerning the distinction between normal and raised concentrations of TPO Abs. The agreement of results (enhanced or normal TPO Ab concentrations in both tests) was shown in 30 sera samples (78.95%), but in residual 8 sera (21.05%) normal TPO Ab concentrations were obtained by Immunotech, and enchanced by Cis biointernational assay. There is no difference in capability of distinction between normal and pathological results between the two tests (χ2=3.484, p gt 0.05). The highest concentration of TPO Ab measured by Cis biointernational assay was not the highest one in Immunotech assay, which might be a reflection of different specificity of antibodies used in two diagnostic tests. Conclusion. TPO Ab concentrations obtained by Cis biointernational and Immunotech assays are very different. In several sera samples, normal concentrations of TPO autoantibodies were obtained by Immunotech assay and enhanced by Cis biointernational assay. The highest value obtained by one is not the highest value measured by another assay we used.Autontitela specifična za tiroidnu peroksidazu autoantitela specifičnih za tiroidnu peroksidazu prevashodno se određuju radi dijagnoze autoimunih bolesti štitaste žlezde. Cilj rada bio je da se uporede koncentracije autoantitela specifičnih za tiroidnu peroksidazu dobijene korišćenjem dva testa: Cis bionternational (Francuska) i Immunotech (Češka Republika). Ispitivanjem je obuhvaćeno 38 ispitanika. Iako su se koncentracije autoantitela specifičnih za tiroidnu peroksidazu u ispitivanim serumima znatno razlikovale i u apsolutnim i u relativnim vrednostima, statističkom obradom rezultata pokazana je direktna korelacija rezultata merenja ova dva testa, sa koeficijentom R=0,6239 (p lt 0,001). Na osnovu analize relativnih vrednosti. pokazana je statistički značajna razlika (p=0,003) između srednjih vrednosti rezultata izmerenih testovima Cis (18,94±37,22) i Immunotech (4,22±8,22). Slaganje rezultata pokazano je u 78,95% seruma, dok je u 21,05% seruma testom Immunotech dobijena normalna, a testom Cis biointernational granična ili povećana koncentracija antitela. Statistički podaci su pokazali da se testovi ne razlikuju po razdvajanju patoloških od normalnih vrednosti (χ2=3,484, p gt 0,05). Iako koncentracije autoantitela specifičnih za tiroidnu peroksidazu izmerene pomoću testova Cis biointernational i testom Immunotech pokazuju značajan stepen korelacije, njihove i apsolutne i relativne vrednosti znatno se razlikuju

Hepatoprotective Effect of Mixture of Dipropyl Polysulfides in Concanavalin A-Induced Hepatitis

The main biologically active components of plants belonging to the genus Allium, responsible for their biological activities, including anti-inflammatory, antioxidant and immunomodulatory, are organosulfur compounds. The aim of this study was to synthetize the mixture of dipropyl polysulfides (DPPS) and to test their biological activity in acute hepatitis. C57BL/6 mice were administered orally with DPPS 6 h before intravenous injection of Concanavalin A (ConA). Liver inflammation, necrosis and hepatocytes apoptosis were determined by histological analyses. Cytokines in liver tissue were determined by ELISA, expression of adhesive molecules and enzymes by RT PCR, while liver mononuclear cells were analyzed by flow cytometry. DPPS pretreatment significantly attenuated liver inflammation and injury, as evidenced by biochemical and histopathological observations. In DPPS-pretreated mice, messenger RNA levels of adhesion molecules and NADPH oxidase complex were significantly reduced, while the expression of SOD enzymes was enhanced. DPPS pretreatment decreased protein level of inflammatory cytokines and increased percentage of T regulatory cells in the livers of ConA mice. DPPS showed hepatoprotective effects in ConA-induced hepatitis, characterized by attenuation of inflammation and affection of Th17/Treg balance in favor of T regulatory cells and implicating potential therapeutic usage of DPPS mixture in inflammatory liver diseases

Murine cytomegalovirus infection induces susceptibility to eae in resistant BalB/c Mice

In contrast to C57BL/6 mice, BALB/c mice are relatively resistant to the induction of experimental autoimmune encephalomyelitis (EAE) after challenge with MOG35–55 peptide. Here, we provide the first evidence that infection with murine cytomegalovirus (MCMV) in adulthood abrogates this resistance. Infected BALB/c mice developed clinical and histological signs similar to those seen in susceptible C57BL/6 mice. In addition to CD4+ cells, large proportion of cells in the infiltrate of diseased BALB/c mice was CD8+, similar with findings in multiple sclerosis. CD8+ cells that responded to ex vivo restimulation with MOG35–55 were not specific for viral epitopes pp89 and m164. MCMV infection favors proinflammatory type of dendritic cells (CD86+CD40+CD11c+) in the peripheral lymph organs, M1 type of microglia in central nervous system, and increases development of Th1/Th17 encephalitogenic cells. This study indicates that MCMV may enhance autoimmune neuropathology and abrogate inherent resistance to EAE in mouse strain by enhancing proinflammatory phenotype of antigen-presenting cells, Th1/Th17, and CD8 response to MOG35–55

Potential preservation of dental pulp stem cells

Dental pulp stem cells (DPSCs) as postnatal stem cells have recently been described. They are clonogenic cells, capable for self-renewal with high proliferative potential. Their multilineage potential and plasticity enables their differentiation into different kind of cells, such as osteoblasts, chondrocytes, adipocytes, muscle cells, neural cells, odontoblasts, cementoblasts and ameloblasts. DPSCs are an important human stem cells source, especially in patients who lost their chance for umbilical cord blood isolation and preservation. As these cells became useful for tissue engineering and cell therapy, proper mode of their preservation also became important. The most important points in the cryopreservation and recovery procedure are: growth phase of harvested cells, number of cells, the proper cryopreservative concentration and serum concentration. The cryopreservation process includes the following general components: harvesting of the cells, addition of cryopreservative, the freezing procedure, the thawing procedure and assessment of the viability prior to transplantation. There is no single and perfect cryopreservation method. Further investigations should be regarding capability of DPSCs and their differentiated cells to recover and restart proliferation, differentiation and new tissue production for therapeutic use after cryopreservation

Docking studies, cytotoxicity evaluation and interactions of binuclear copper(ii) complexes with s-isoalkyl derivatives of thiosalicylic acid with some relevant biomolecules

The numerous side effects of platinum based chemotherapy has led to the design of new therapeutics with platinum replaced by another transition metal. Here, we investigated the interactions of previously reported copper(II) complexes containing S-isoalkyl derivatives, the salicylic acid with guanosine-5′-monophosphate and calf thymus DNA (CT-DNA) and their antitumor effects, in a colon carcinoma model. All three copper(II) complexes exhibited an affinity for binding to CT-DNA, but there was no indication of intercalation or the displacement of ethidium bromide. Molecular docking studies revealed a significant affinity of the complexes for binding to the minor groove of B-form DNA, which coincided with DNA elongation, and a higher affinity for binding to Z-form DNA, supporting the hypothesis that the complex binding to CT-DNA induces a local transition from B-form to Z-form DNA. These complexes show a moderate, but selective cytotoxic effect toward colon cancer cells in vitro. Binuclear complex of copper(II) with S-isoamyl derivative of thiosalicylic acid showed the highest cytotoxic effect, arrested tumor cells in the G2/M phase of the cell cycle, and significantly reduced the expression of inflammatory molecules pro-IL-1β, TNF-α, ICAM-1, and VCAM-1 in the tissue of primary heterotopic murine colon cancer, which was accompanied by a significantly reduced tumor growth and metastases in the lung and liver

The effect of propylthiouracil on function of phagocytic peripheral blood cells in persons with thyroid hyperfunction

Introduction. It is known that hyperthyroidism as well as thyrosuppressive therapy can influence the cells of immunological system. Objective. To examine the function of phagocyte cells in persons with hyperthyroidism and to examine if propylthiouracil (PTU) influences this function. Method. The study included 15 patients with hyperthyroidism and 10 healthy persons. The parameters of phagocytic activity of mononuclear and polymorphonuclear leucocytes were tested by method of ingestion of particles of inactivated yeast labeled with neutral-red. Results. It was demonstrated that patients with hyperthyroidism, before the onset of therapy as well as 14 days after introduction of PTU, had decreased number of leucocytes (before PTU: 6.7±3.2Ч109/l, after PTU: 6.1±2.0Ч109/l and control: 8.0±1.6Ч109/l; p=0.039), PMN leucocytes (before PTU: 3.9±2,4 Ч109/l, after PTU: 3.5±1.6Ч109/l and control: 4.8±0.9Ч109/l; p=0.037) and number of phagocyte PMN cells (before PTU: 0.9±0.9Ч109/l, after PTU: 0.9±0.7Ч109/l and control: 1.3±0.6 Ч109/l; p<0,05), but they had increased index of phagocytosis (before PTU: 2.0±0.2, after PTU: 1.9±0.2 and control: 1.7±0.2; p=0.029), while capacity of phagocytosis remained unchanged (before PTU: 1.9±1.7Ч109/l, after PTU: 1.6±1.9Ч109/l and control: 2.4±1.4Ч109/l; p>0.05). The number of mononuclear leucocytes and parameters of phagocytic activity of mononuclear phagocytes in persons with hyperthyroidism did not change significantly in comparison with the control group. Conclusion. Patients with hyperthyroidism had decreased number of leucocytes, PMN leucocytes and number of phagocyte PMN cells, and increased index of phagocytosis, while capacity of phagocytosis remained unchanged. The number and parameters of phagocytic activity of mononuclear leucocytes did not change. PTU therapy had no effect on the examined parameters