[TREATMENT AND PREVENTION OF METASTASIS TO THE PRE- AND PARATRACHEAL LYMPH NODES IN CANCER OF THE LARYNX WITH SPREAD TO THE SUBGLOTTIC SECTION].

Based on the specificity of the subglottic cancer, the basic method of treatment is surgical, because many years of experience have shown the ineffectiveness of chemotherapy and the low effectiveness of radiation therapy. We have developed and introduced in practice the method of extended-combined laryngectomy is effective not only in terms of treating the underlying disease, but also for preventive purposes, reducing the risk of metastases in pre- and paratracheal lymphatic zones. The method of extended-combined laryngectomy includes mobilization of the larynx and its removal at level V or VI of the tracheal rings, lymph node dissection at level VI, both on the affected side and prophylactically from the opposite side, as well as removal of either the thyroid gland on the side of the lesion subglottic cancer, or at its circulatory location total removal of the gland. Despite the ongoing controversy over such a scale of operations, the effectiveness of extended-combined laryngectomy remains irrelevant, as indicated by a decrease in the frequency of metastasis recurrence from 65.5% to 15.3% with cancer of the subglottic localization and from 63.2% to 25.8% in the case of the spread of the tumor to the subglottic section.

ALTERATION IN THE CYTOKINE SECRETION PATTERN IN T CELLS OF PATIENTS WITH CYSTIC FIBROSIS CAUSED BY DNA METHYLTRANSFERASE INHIBITOR 5-AZACITIDINE.

Cystic fibrosis (CF) is the autosomal-recessive disorder caused by mutation in the cystic fibrosis transmembrane conductance regulator gene (CFTR). The Airway inflammation plays a central role in the progression of CF disease. Cystic fibrosis characterized by the overproduction of the pro-inflammatory cytokines and reduced expression of anti-inflammatory cytokines. Although the mechanisms of abnormal cytokine expression is still poorly understood, altered epigenetic regulations in T cells might contribute. In the present study we examined the expression of IFN-γ and IL-10 by CF T cells prior to and following 5-azaC treatment. In addition we investigated DNMTs levels in nuclear extracts of CD4+ T cells derived from CF and non-CF individuals. Seven CF patients (age: 5-12 years) were included in the study and compared to six age-matched healthy subjects (age: 6- 13 years). CD4+ T cells were isolated from PBMC using CD4 MicroBead kit (Miltenyi Biotec GmbH) and were cultured in RPMI 1640 medium at 37°C with 5% CO2, in presence or absence of 5-azacytidine. Concentrations of IL-10 and γ-INF in CD4+ T Cells were measured by ELISA (eBoiscience, san Diego, CA, USA). In our study we showed that 5 Azacytidine alters nuclear levels of DNMT 3a as well as modulates cytokine levels in CD4+ T cells derived from CF patients. After 5-azaC treatment secretion of IFN-γ was significantly decreased in CF T cells, while amount of IL-10 was elevated by ~2.5 times compared to untreated controls (P<0.05). In summary, data presented in this report demonstrates that epigenetic mechanisms such as DNA methylation may be considered as a one of the potential therapeutic target in a treatment of Cystic Fibrosis.

mGluR1 interacts with cystic fibrosis transmembrane conductance regulator and modulates the secretion of IL-10 in cystic fibrosis peripheral lymphocytes.

Cystic fibrosis (CF) is caused by the mutations in the gene encoding the cystic fibrosis transmembrane conductance regulator (CFTR) chloride channel. CFTR dysfunction in T cells could lead directly to aberrant immune responses. The action of glutamate on the secretion of IL-8 and IL-10 by lymphocytes derived from healthy subjects and cystic CF patients, as well as the expression of metabotropic glutamate receptor subtype 1 (mGluR1) in the membrane fractions of lymphocytes was investigated. Our results have shown that CF-derived T-cells in the presence of IL-2 produce more IL-8 and IL-10, than T-cell from healthy control. However, only in normal lymphocytes a significant increase (144%) in the IL-10 secretion during exposure to high concentration of glutamate (10(-4)M) was detected. Glutamate-dependent secretion of IL-10 was not inhibited either by NMDA-receptor (NMDAR), or by AMPA-receptor (AMPAR) antagonist. Only mGluR1 antagonist, LY367385, strongly decreases the production of IL-10. Furthermore, the content of mGluR1, as well as cystic fibrosis transmembrane conductance regulator-associated ligand (CAL), Na(+)/H(+) exchanger regulatory factor 1 (NHERF-1), was analyzed in plasma membrane of lymphocytes after immunoprecipitation of CFTR. We have found that normal, non-mutated CFTR, as well as mutated forms of CFTR were associated with metabotropic mGluR1, but the level of surface exposed mGluR1 in CF-lymphocytes was much lower than in normal cells. Besides, our results have shown that normal, non-mutated CFTR, as well as mutated forms of CFTR were associated with NHERF-1 and CAL; however in lymphocytes with CFTR mutation the amount of cell-surface expressed CFTR-CAL complex was greatly decreased. We have concluded that CFTR and mGluR1 could compete for binding to CAL, which in turn downregulates the post-synthetic trafficking of mGluR1 and decreases the synthesis of IL-10.

Glutamate decreases the secretion of IL-10 by peripheral blood lymphocytes in persons with autoimmune thyroiditis.

Human T lymphocytes expose ionotropic and metabotropic glutamate receptors, which control immune responses, cell activation, maturation, and death. Several cytokines release during inflammation which identification may have important physiological and clinical implications. Main biological function of IL-10 is limitation and termination of inflammatory responses and the regulation of differentiation and proliferation of several immune cells. Various inflammatory molecules regulated the secretion of IL-8 and IL-10, but the action of glutamate on the biosynthesis of cytokines is unknown. We have found that in peripheral blood lymphocytes glutamate at the concentrations within normal plasma levels (1 x 10(-5) M), as well as at lower concentration (0.3 x 10(-6) M) changes the secretion of immunosuppressive cytokine IL-10, whereas synthesis of proinflammatory chemokine, IL-8 did not changed significantly. Moreover, our results have shown that peripheral blood lymphocytes from patients with autoimmune thyroiditis release less IL-10 at both concentration of glutamate than peripheral blood lymphocytes from healthy persons. These data suggest that glutamate decrease the secretion of IL-10 by peripheral blood lymphocytes, especially in patients with autoimmune thyroiditis that may be responsible for prolongation of inflammation.

[The action of dextrorphan and sigma ligands on the spontaneous secretion of interleukins by jurkat cell line].

Sigma receptor was demonstrated to have at least two subtypes, mediating pharmacological effects of various preparations including psychoactive, neuroleptic, cardioprotector, anti-inflammatory, immunosuppressive compounds and several steroid hormones. The stimulation of sigma receptor induces transient increase of intracellular calcium and amplifies signals from different stimuli. Pentazocine, SKF 10 047, dextrorphan, and other sigma ligands including phencyclidine and haloperidol were investigated for their potential immunoregulatory properties. We have found, that pentazocine, SKF 10 047, dextrorphan reduce spontaneous secretion of IL-8, IL-6 and IL-10 and selectively changes synthesis of IL-4 by Jurkat human T lymphocyte cells lines. Dextrorphan significantly enhanced, pentazocine, haloperidol and phencyclidine had no effect, while SKF 10 047 suppressed production of IL-4. Spontaneous secretion of IL-4 and IL-8 correlates with synthesis of nitric oxide, suggesting that NO and transitory S-nitrosylation of up-stream proteins participate in the sigma ligand dependent expression of IL-4 and IL-8 genes.