The Involvement of Semaphorins in the Pathogenesis of Skin Diseases.

Semaphorins belong to a group of membrane and secretory proteins that act as ligands for several receptor families and are involved in modulating cell signaling pathways. They bind multimeric receptor complexes on the cell membrane to exert their effects and initiate unique intracellular signal transduction cascades. These proteins can influence several processes that are very important for cell function, such as cell division and differentiation. Semaphorins are involved in cell migration, apoptosis, cell adhesion, aggregation, and numerous immune processes due to their immunoregulatory effects. Semaphorins are expressed in keratinocytes, which is why they have become a target for studies on the pathogenesis of skin diseases. Most studies to date on the role of semaphorins in the pathogenesis of skin diseases have been carried out in cellular or animal models, and there are few clinical studies evaluating the role of semaphorins in the pathogenesis and therapy of skin diseases. In this narrative review, we summarized the current state of knowledge on the role of semaphorins in the pathogenesis of skin diseases and their potential importance as targets for therapy. We also tried to present the key findings and weaknesses of previous research in this field. The novelty of this article lies in the comprehensive presentation of the role of semaphorins in the pathogenesis of skin diseases, including the results of studies on cell cultures and animal models, elucidating the mechanisms and signaling pathways through which semaphorins affect the development of skin diseases, as well as on the presentation of the results of existing clinical trials evaluating the role of semaphorins in the pathogenesis of skin diseases, and as potential therapeutic targets.

The Impact of Regional Maximum Tolerated Interlesion Distance on the Long-Term Ablation Outcomes in Ablation Index Guided Pulmonary Vein Isolation for Atrial Fibrillation.

BACKGROUND: An adequate interlesion distance (ILD) applied during point-by-point pulmonary vein (PV) isolation for atrial fibrillation (AF) has never been established. We hypothesized that maximum tolerated ILD may differ between PV regions and may influence long-term ablation outcomes. METHODS: A total of 260 AF patients underwent PV isolation with 3D electroanatomical platform. Postablation, ILD values were classified into 5 groups (6-5.5 mm, 5.5-5.0 mm, 5.0-4.5 mm, 4.5-4.0 mm and <4.0 mm); the number of tags in each group was calculated and correlated with postablation AF recurrence (AFR). All measurements were performed globally for the entire encirclement around each individual PV and regionally for designated PV anatomic segments. RESULTS: Single-procedure freedom from AF was 81% for paroxysmal and 66% for persistent AF at 12 months. Global analysis showed that AFR was not related to median ILD nor the number of lesions within each ILD tag group for any PV. Segmental analysis showed that AFR was not related to median ILD. However, the presence of tags from the 5.5-6.0 mm ILD group located on the posterior aspect of right upper PV (RUPV) correlated with AFR. This was confirmed in a multivariable logistic regression model. CONCLUSIONS: Maximum tolerated ILD of 6.0 mm translated into well-accepted ablation results. However, the study suggests that it may be inadequate at the posterior aspect of RUPV.

Immunotherapy of lung adenocarcinoma patient with Peptide-pulsed dendritic cells: a case report.

Immunotherapy with ex vivo generated dendritic cells (DCs) is reported to be of low toxicity and of diverse effectiveness in cancer treatment. The synthetic antigens are frequently used for immunotherapy especially for patients with stable disease after prior treatment. We described the effect of peptide-loaded DCs-based immunotherapy on patient with recurrent surgically resected adenocarcinoma with bronchoalveolar feature with co-existing of Takayasu arteritis and chronic hepatitis B. In January 2010, 61-year-old patient received subcutaneously four bi-weekly vaccinations of DCs loaded with MUC1 and MAGE-3 epitopes. Additionally, he received three bi-weekly booster vaccinations after 7 months from the first course of immunotherapy. Delayed-type hypersensitivity test was positive only for MAGE-3 antigen. The evidence expansion of MAGE-3-specific CD8(+) cells after first vaccination and after third vaccination during boosters injections was observed (from 0.08% before vaccination to 0.5% after first vaccination; from 0.05% before booster vaccination to 0.24% after third injection). Computed tomography scans performed after first course and after booster course of vaccination until April 2011 did not shown any presence of lung tumour or metastases. Based on clinical factors (no completed wedge-resection and recurrent character of cancer) as well as on the presence of the tumour-antigen-specific immunological response, we could speculated that immunotherapy prolonged disease free-survival in our patient. Over 16 months from first vaccination, the patient remains without symptoms of cancer.

[Evaluation of Th1/Th2 lymphocyte balance and lipopolysaccharide receptor expression in asthma patients]. / Ocena równowagi limfocytów Th1/Th2 oraz ekspresji receptorów dla lipopolisacharydu u chorych na astme.

INTRODUCTION: An increase in the number of asthma patients which has recently been observed depends on their place of residence and their occupation. This suggests that both external factors and genetic predispositions affect the development of the disease. The contact with bacterial lypopolysaccharide (LPS) may suppress the development of asthma among rural inhabitants. The mechanism of LPS effect most probably consists in the activation of macrophages and granulocytes by TLR4 and CD14 receptors for the production of cytokines, which affect Th1/Th2 balance. The objective of the study was the evaluation of CD14 and TLR4 expression on mononuclear cells and the analysis of Th1/Th2 balance in peripheral blood among asthma patients. MATERIAL AND METHODS: The study group covered 22 patients with bronchial asthma (mean age 45 +/- 15), and was conducted by the method of flow cytometry with the use of fluorochrome-labelled monoclonal antibodies. CD14 and TLR expression was assessed in peripheral blood monocytes. Th1/Th2 balance was determined by the measurement of intracellular IL-2, IFN-gamma, IL-4 and IL-10 expression in T-helper cells after culture with the stimulation of cytokine production. RESULTS: A negative correlation was noted between TLR4 expression and the percentage of Th2 lymphocytes, while a positive correlation was observed between expression of TLR4 and percentage of Th1 cells. No relationship was found between CD14 expression on monocytes and the percentage of Th1 and Th2 lymphocytes. CONCLUSIONS: An increased percentage of lymphocytes with TLR4 expression is associated with the change in Th1/Th2 balance in favour of Th1 lymphocytes in asthma patients.

Phenotype of dendritic cells generated in the presence of non-small cell lung cancer antigens - preliminary report.

Therapeutic outcomes of definitively treated non-small-cell lung cancer (NSCLC) are unacceptably poor. It has been proposed that the manipulation of dendritic cells (DCs) as a "natural" vaccine adjuvant may prove to be a particularly effective way to stimulate antitumor immunity. Presently, there is no standardized methodology for preparing vaccines and many questions concerning the optimal source and type of antigens as well as maturation state and activity of DCs are still unsolved. The study population comprised of ten patients with histologically confirmed NSCLC (mean age: 67.63 +/- 6.15 years). Resected small tumor pieces were placed in tissue culture dishes containing different growth factors in order to obtain pure cancer cells. Seven days after the operation, the PBMC were collected and monocytes were purified by the adherence to culture dishes. Monocytes were cultured in RPMI 1640 medium supplemented with 10% of autologous plasma in the presence of rhIL-4 and rhGM-CSF to generate immature autologous (DCs). TNF-alpha with or without tumor cells' lysate were added to maturation of DCs. After 7 days of culture, DCs were harvested and the expression of CD1a, CD83, CD80, CD86 and HLA-DR antigens were analyzed by flow cytometry. We discovered higher (p=0.07) percentage of semimature DCs in tumor cell lysate culture in comparison with TNF-alpha culture (21.22 +/- 16.82% versus 11.27 +/- 11.64%). The expression of co-stimulatory and maturation markers (CD86, CD83 and HLA-DR) was higher on DCs from the culture with tumor cell lysate compared with TNF-alpha culture as a control. Specimen of NSCLC's culture prepared in this way could generate differences in DCs phenotype, which may have an influence on the therapeutic and protective antitumor immunity of the vaccine. Our research seems to be the next step in the development of DC-based vaccine. We are going to continue the investigation to start the preparation of a pattern of immunological vaccine against lung cancer.

Tumor necrosis factor receptors (TNFRs) on T lymphocytes and soluble TNFRs in different clinical courses of sarcoidosis.

INTRODUCTION: The release of tumor necrosis factor (TNF-alpha) is increased in sarcoidosis patients. TNF-alpha exerts its effect by binding to specific cell surface receptors. There are only fragmentary data concerning the expression of tumor necrosis factor receptors (TNFRs) on bronchoalveolar lavage fluid (BALF) and peripheral blood (PB) lymphocytes. The aim of the study was to evaluate TNFRI (CD120a) and TNFRII (CD120b) expression on T cells and the level of soluble TNFRs in specimens of patients with different clinical manifestation and clinical outcome of sarcoidosis. MATERIAL AND METHODS: We examined 49 patients with newly diagnosed pulmonary sarcoidosis. TNFRI and TNFRII density on CD4+ and CD8+ BALF and PB cells surface was estimated using monoclonal antibodies and a flow cytometry technique. The level of TNFRs in PB serum and BALF cell culture supernatant (CCS) was measured using ELISA. Immunological analyses were also performed on PB samples collected from 10 healthy volunteers. RESULTS: The level of soluble TNFRI (sTNFRI) in PB serum was similar in sarcoidosis patients and healthy subjects, whereas the concentration of sTNFRII in serum was significantly higher in the sarcoidosis group (P<0.001). Patients without acute symptoms of sarcoidosis, patients with radiological stage II/III as well as patients with further disease progression showed a tendency to higher levels of sTNFRs in PB serum and lower levels of sTNFRs in BALF CCS compared to Löfgren syndrome and radiological stage I subjects, and patients with spontaneous resolution of sarcoidosis. More than 80% of BALF and PB lymphocytes of sarcoidosis patients expressed both CD120a and CD120b antigens. The percentage of double-positive CD4+CD120a+ and CD4+CD120b+ cells in PB was significantly higher (P<0.005) in sarcoidosis patients than in healthy subjects. The highest percentage of CD4+CD120a+ and CD4+CD120b+ lymphocytes in BALF was determined in patients with acute disease, and in PB of patients with further spontaneous improvement. CONCLUSION: The evaluation of sTNFRs and TNFRs expression on T-helper cells may be useful in the estimation of sarcoidosis activity.

The clinical significance of interleukin 18 assessment in sarcoidosis patients.

INTRODUCTION: Sarcoidosis is a multisystemic disease of unknown etiology characterized by the formation of immune granulomas in involved organs. The cytokine profile in inflamed lesions of sarcoidosis is mainly determined by T helper 1 (Th1) cells. Interleukin 18 (IL-18) is primarily a monocyte/macrophage-derived cytokine. IL-18 has been recently identified as an IFNgamma-inducing factor. The cytokine plays an important role in the induction of Th1 response and it may be responsible for sarcoidosis progression. The aim of the study was to assess the usefulness of IL-18 estimation in the sarcoidosis diagnosis and the disease course prognosis. MATERIAL AND METHODS: The diagnosis of sarcoidosis was established in 88 patients (the mean age of 38.1+/-10.8 years). We measured IL-18 level in plasma and bronchoalveolar lavage fluid (BALF) cell culture supernatant (CCS) using the enzyme-linked immunoassay technique (ELISA). We also performed the flow cytometric analysis of BALF lymphocyte phenotype. Statistica 5.0 and non-parametric tests: the Mann-Whitney U-test and the Spearman correlation test, were used for statistical analysis. RESULTS: The patient group consisted of 55 subjects without acute symptoms of sarcoidosis, 14 patients with acute Löfgren syndrome and 19 subjects with Löfgren syndrome in the past. Lung hilar lymphadenopathy was diagnosed in 49 patients and lung interstitial changes in 39 subjects. After 6-month-observation, 49 patients were in remission, 20 subjects manifested persistent disease and 19 patients had sarcoidosis progression. Plasma IL-18 level was significantly (P<0.0001) higher in sarcoidosis patients (383+/-250pg/ml) than in control subjects (146+/-72pg/ml). Plasma IL-18 level was similar both in subjects with Löfgren syndrome and in other patients. However, IL-18 level in BALF CCS was significantly (P<0.05) lower in Löfgren syndrome patients than in subjects without acute manifestation of the disease. The highest IL-18 level in plasma was found in patients with disease progression, in subjects with lung interstitial changes and in patients with extrapulmonary manifestation of the disease. We observed a positive correlation between plasma IL-18 level and the percentage of BALF lymphocytes (R=0.202, P=0.06) as well as the percentage of activated HLA DR+T cells (R=0.23, P<0.05). There was a negative correlation between the IL-18 level in BALF CCS and the percentage of BALF CD3-positive and CD4-positive lymphocytes (R=-0.27, -0.23, P<0.05). CONCLUSION: IL-18 may play a significant role in the prolongation of sarcoidosis course. Its estimation may become a good prognostic factor, which should be analyzed together with other factors useful in sarcoidosis monitoring.